• Volume 38,Issue 6,2014 Table of Contents
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    • The effect of acutehandling stress on the stress-related neuroendocrine factor in Coilia nasus

      2014, 38(6):803-812. DOI: 10.3724/SP.J.1231.2014.49029

      Abstract (2447) HTML (0) PDF 3.08 M (1564) Comment (0) Favorites

      Abstract:The present study aimed to investigate the changes in mRNA levels for the stress-related neuroendocrine factor in response to the acute handling stress in Coilia nasus.The radioimmunoassay(RIA)and chemical luminal immunoassay(CLIA)were introduced to detect the cortisol levels in plasma and head-kidney.Fish in stress group showed a significant elevation of cortisol concentration in both plasma(P<0.01)and head-kidney(P<0.05).The plasma cortisol concentration and head-kidney cortisol concentration have risen by 56.48% and 49.68%,respectively,suggesting that the acute stress response was instantly induced after handling.Moreover,the partial sequences of C.nasus corticotropin-releasing hormone(CRH),urotensin I(UI)and proopiomelanocortin(POMC)were cloned by homology cloning,and C.nasus brain CRH,POMC and UI mRNA levels were determined by real-time quantitative PCR(RT-qPCR).The CRH(P<0.01)and the POMC(P<0.05)mRNA expressions were significantly suppressed after a half-hour handling stimulation.But there was no significant change in brain UI mRNA level.Taken together,the acute stress response in C.nasus could be regulated by the stress-related neuroendocrine factor,including cortisol,CRH,UI and POMC,via the hypothalamic-pituitary-interrenal stress axis,which gave us a valuable insight into the C.nasus stress system,and could help developing the stress relief strategy for C.nasus.

    • Histopathological and pathogeny preliminary research on mulberry heart of Siberian sturgeon(Acipenser baeri)

      2014, 38(6):904-911. DOI: 10.3724/SP.J.1231.2014.48967

      Abstract (2158) HTML (0) PDF 8.83 M (2457) Comment (0) Favorites

      Abstract:In recent years,a kind of disease broke out in Siberian sturgeon breeding base of Beijing.The main symptoms of diseased Siberian sturgeon were that the heart surface was observed to have a large number of rice grain-like,purple-brown cystic neoplasm.The whole heart looks like mulberries shape.The mortality rate were as high as 60% or more.Sick fish were about one year old.Water temperature was around 27℃ or so.In order to diagnose and preliminarily discuss the cause on heart surface cystic neoplasm of Siberian sturgeon,the histopathologic changes of the diseased Siberian sturgeon and the morphology of the pathogen were observed under optic and electronic microscopes.The results show that the heart surface of sick Siberian sturgeon can be observed in a large of rice grain-like,purple-brown cystic neoplasm.The heart was like mulberries shape.The epicardial mesothelial cells and connective tissue were between obvious hyperplasia.Neoplasm was formed.Epicardial lesions were dynamic process.Epicardial lesions were becoming more severe from mild.The main show were epithelium by flat to cuboidal epithelium.The cyst walls were gradually thickening and differentiating,so that the increasing cysts lined become more.The cyst cavity had a number of undifferentiated spherical,spindle and so on the different mature degrees of blood cells and vascular tissue,and infiltration with a lot of free or round state distribution inflammatory cells.The whole lesions looked like a kind of tumor type of malignant hyperplasia,myocardial cells were swelling,fractured,dissolved and necrotic.The liver lesions were hepatic steatosis and infiltration of inflammatory cells.Renal interstitium and spleen lymphocytes increased.Gill lamellae were observed between the large number of bacterial invading epithelial cells;the epithelial cells of gill lamellae were severe swelling and hyperplasia.Under transmission electronic microscope,a large number of bacteria were observed in the heart,liver,spleen and kidney.The microscopic and ultrastructure changes speculate that its etiology may be associated with trace elements and the bacteria are not the direct cause of the disease.To provide basic information for the prevention and treatment,clinical diagnosis and pathogenesis and related research,the hispathological changes were more systematically researched on Siberian sturgeon mulberry heart disease.

    • Isolation of antialgal substances from Enteromorpha prolifera and its growth inhibition on the red-tide microalgae

      2014, 38(6):813-819. DOI: 10.3724/SP.J.1231.2014.49010

      Abstract (2236) HTML (0) PDF 5.73 M (1762) Comment (0) Favorites

      Abstract:Antialgail activity extractions EPME(methanol extracts of Enteromorpha prolifera)were separated by liquid-liquid extraction and silica gel column chromatography to isolate and identify antialgal activity compounds in EPME.By liquid-liquid extraction,the four kinds of components(Ⅰ,Ⅱ,Ⅲ and Ⅳ)were obtained.Their yield was 29.0%,19.0%,12.7% and 5.45%,respectively,but only component Ⅱ has only obvious inhibiting activity.The inhibitory effect of Karenia mikimitoi,Skeletonema costatum and Alexandrium tamarense by component Ⅱ(1.0 g/L)was 43.4%,44.9% and 54.3% on Day 12,respectively.The five kinds of fractions(Ⅱ-A,Ⅱ-B,Ⅱ-C,Ⅱ-D and Ⅱ-E)were obtained by silica gel column chromatography,respectively.The fraction B(Ⅱ-B)and fraction C(Ⅱ-C)inhibited significantly the growth of all test microalgae,and these two fractions caused cells cavities and pieces.Further,these two fractions(Ⅱ-B andⅡ-C)were analyzed by wavelength scanning,compound identification test and silica gel GF254 thin-layer chromatography.This paper identified containing lactone and coumarins in them and determined the appropriate developing agent by silica gel GF254 thin-layer chromatography for subsequent separation and purification.This paper provided solid experiment foundation for further study.

    • Primary investigations on the photosynthesis and respiration of Macrocystis pyrifera

      2014, 38(6):820-828. DOI: 10.3724/SP.J.1231.2014.49062

      Abstract (2230) HTML (0) PDF 17.68 M (1346) Comment (0) Favorites

      Abstract:For clarifying the development and ecological adaptability of juvenile sporophytes of Macrocystis pyrifera,the effects of photoperiod(10 L:14 D,12 L:12 D,14 L:10 D,16 L:8 D)on gametophyte clone seed-rearing of M.pyrifera were studied.And,the impacts of light intensity,temperature,nutrients concentration on the growth,net photosynthetic rate(Pn),dark respiratory rate(Rd)of sporophytes(0.5-1 cm)of Macrocystis pyrifera were investigated by single factor test and orthogonal experiment.The major results included:(1)Large numbers of gametophytes developed into sporophytes at long photoperiod(14 L:10 D,16 L:8 D).(2)The highest value for relative growth rate(RGR)and Pn were observed at 70 μmol/(m2·s)light intensity,13℃ temperature and 2 mg/L nitrogen concentration(N:P=10:1,the same below).The growth of sporophytes was inhibited when exposure to 90 μmol/(m2·s),RGR of which was significantly lower than other tests.RGR at 10℃ was significantly lower than that at 13℃(P<0.05).And RGR significantly decreased when exposure to 8 mg/L nitrogen concentration.(3)Pn and Rd presented the rising trend with increased nutrients concentration.Pn of 0.2 mg/L group was significantly lower than that at 6 mg/L and 8 mg/L group.Rd significantly different compared with other groups while cultured at 0.2 mg/L(P<0.05).(4)Pn decreased with rising of temperature while cultured at the same light intensity.At the range of 40-80 μmol/(m2·s),Pn was significantly different between 10,13 group and 16,19℃ group.According to orthogonal experiment,the optimum light intensity,temperature and nitrogen concentration for cultivation of sporophytes of M.pyrifera were 40-70 μmol/(m2·s),13℃ and 2 mg/L.Our results provide guidance for the optimization of artificial breeding technologies for M.pyrifera.

    • The burst-coast swimming behavior of grass carp (Ctenopharyngodon idellus)during fast-start

      2014, 38(6):829-834. DOI: 10.3724/SP.J.1231.2014.49057

      Abstract (2344) HTML (0) PDF 2.48 M (1507) Comment (0) Favorites

      Abstract:To learn capability of grass carp(Ctenopharyngodon idellus)to cross velocity barrier,laboratory experiments were conducted to test burst-coast swimming performance of grass carp through startle response.In this study,the tested fish were divided into three groups according to their body length:(8.47±0.73)cm(small-size),(17.93±1.27)cm(mid-size) and (51.24±3.24)cm(large-size).The experiments were carried out in an environment with natural light and water temperature of(20.00±1.50)℃.The results showed that the highest absolute sprinting speeds of fish were(1.449±0.424)m/s(achieved in 0.294 s),(2.359±0.434)m/s(0.294 s) and (2.899±0.457)m/s(0.378 s)for small-size,mid-size fish and large-size fish respectively; the relative sprinting speeds were(17.099±5.009)BL/s,(13.156±2.418)BL/s,and(5.659±0.891)BL/s,respectively.Fish were observed to slow down to coast by keeping their body straight(no tail-beat)after fish reached their highest sprinting speeds.Absolute sprinting speed of grass carp was found to increase with body length(P>0.05),while relative sprinting speed decreased as body length increased.Relative sprinting speed of small-size fish was significantly higher than that of large-size fish(P<0.05).Absolute acceleration of fish was not affected by size(P>0.05); while the absolute coast deceleration of large-size fish was significantly higher than those of smaller size fish(P<0.05); and the relative burst acceleration and relative coast deceleration of large-size fish were both significantly lower than that of small-size fish(P<0.05).

    • The influence of stocking density on the growth performance of juvenile Russian sturgeon(Acipenser gueldenstaedti)in flowing water cultivation

      2014, 38(6):835-842. DOI: 10.3724/SP.J.1231.2014.49104

      Abstract (2433) HTML (0) PDF 1.77 M (1811) Comment (0) Favorites

      Abstract:Nowadays,some countries in Europe,Asia,and North America have developed the techniques for sturgeon artificial breeding according to the conservation acts.Russian sturgeon(Acipenser gueldenstaedti)is one of the rare and valuable fish species,which has become one of the popular sturgeon culture species in China.It has been demonstrated that people tend to get more benefit by increasing stocking density,which may suppress fish growth due to the decreasing of food consumption efficiency and the alteration in metabolic rate induced by crowding stress associated with stocking densities.This study was undertaken to investigate the influence of different stocking density on the growth performance of juvenile Russian sturgeon.Three triplicate groups of juveniles[initial body weight:(29.7±1.32)g]were reared in nine square concrete ponds(4.4 m×4.4 m×0.45 m)at three stocking densities[2.5(SD1),3.6(SD2),4.7 kg/m3(SD3)]for 90 days.The results showed that:the weight gain(GBW),length gain(GBL),specific growth rate(SGR)and food conversion ratio(FCR)of the fish at low density(SD1)were(362.01±15.87,55.88±4.77,1.79±0.03,114.95±4.52),respectively,which were significantly higher than those in high density(SD3)(272.30±2.74,46.34±6.22,1.53±0.02,94.49±1.96)(P<0.05).The mortality and greater variation in size and growth of the juvenile fishes in SD3 were significantly higher than those in SD1(P<0.05); Meantime,the protein and fat content decreased while the moisture and ash content increased along with the density increasing,but there is no significant difference(P>0.05).These results suggested that high density has a significant negative impact on the growth preformance of juvenile Russian sturgeon,and the growth suppression caused by high stocking density might be associated with the crowding stress and reductions in both food consumption and food conversion efficiency.

    • Preliminary study on reproductive biology of summer spawning stock of Illex argentinus in the southwestern Atlantic Ocean

      2014, 38(6):843-852. DOI: 10.3724/SP.J.1231.2014.48931

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      Abstract:Illex argentinus is a neritic-oceanic squid species,not only supporting the most important world cephalopod fishery in the terms of landing volumes,but also playing an important role in the southwestern Atlantic Ocean ecosystem owing to its high abundance.However,like other annual lifespan Ommastrephid species,it has been found that the recruitment abundance was closely related to the reproductive success the year before.So,in this study,148 females and 99 males of summer spawning stock of I.argentinus were randomly sampled from the catch of Chinese jigger fleet during the fishing season from December 2012 to March 2013,in the high sea of southwestern Atlantic Ocean,and their reproductive biology was studied,in order to fully understand the important reproductive aspects of this species biology.Results showed that the mantle length of this stock during the fishing season was in the range of 172-252 mm for females and 164-248 mm for males.According to the sampled months,the proportion of females decreased monthly,from 84.21% in December to 27.03% in March,but inversely for males,which had the smallest proportion of 15.79% in December,and then increased to the largest proportion of 72.97% in March.Among the females,more than 30% of individuals were mature and some of them were spawning.However,the mature males were in the proportion of more than 60%,nearly twice of that of females,and some of them had mated.With the gonads development,both of females and males grew gradually and attained their largest mantle length and body weight at the functionally maturity stage.The mantle length of females was longer than that of males according to the maturity stages,whereas the body weight of females was lighter than that of males,and the possible reasons were discussed.Moreover,the gonadosomatic index(GSI)for both sexes,nidamental gland index(NGI)for females and spermatophoric complex index(SCI)for males gradually increased with growth,even though the males' SCI was at a same low level during the immature phase.The females' GSI and NGI were in the range of 0.001 2-0.31,with a mean value of 0.074 and 0.003 1-0.55,with a mean value of 0.29,respectively,and both indices attained their greatest value at the functionally maturity stage.The males' GSI ranged from 0.007 5 to 0.084 with a mean value of 0.038,and SCI ranged from 0.000 79 to 0.035 with a mean value of 0.012.These two indices attained their greatest value when the individuals were being mated.

    • Ecosystem service value assessment for an oyster farm in Dapeng Cove

      2014, 38(6):853-860. DOI: 10.3724/SP.J.1231.2014.49101

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      Abstract:In this study,we assessed the ecosystem service value of the oyster farm in Dapeng Cove,Daya Bay,using the methods and procedures of Millennium Ecosystem Assessment and Technical Directives for Marine Ecological Capital Assessment.Results indicated that,the total ecosystem service value(ESV)of the oyster farm in 2012 was 3 460.52×104 RMB,and the unit ESV was 17.30×104 RMB/hm2.The aquaculture production service value(3 158.00×104 RMB)accounted for the largest proportion of the ESV(91.26%); besides,the recreation service value(124.00×104 RMB)and scientific research service value(71.52×104 RMB)accounted for comparatively high proportions(3.58% and 2.07%,respectively); the rest,such as oxygen generation value(31.34×104 RMB),climate regulation value(61.06×104 RMB)and waste treatment value(14.60×104 RMB)accounted for lower proportions of the ESV.However,for the year 2013,the total ESV of this area decreased to 814.10×104 RMB,and the unit ESV decreased to 4.07×104 RMB/hm2.In the composition of the ESV,the aquaculture production service value(540.00×104 RMB)was much lower than that of the former year,but it still accounted for the largest proportion(66.33%)of the ESV; the oxygen production service value(31.34×104 RMB),recreation service value(124.00×104 RMB)and scientific research service value(71.52×104 RMB)were consistent with the previous year,however,their proportions increased to 3.85%,15.23% and 8.79% of the ESV,respectively; the regulating service(including climate regulation and waste treatment)value decreased sharply during this year,however,it accounted for comparatively higher proportion of the ESV than that of 2012.We concluded that the degradation in the ESV was due to decrease of aquaculture production service,which was mainly caused by high stocking intensity and environmental degradation.In conclusion,the oyster culture promotes the ESV of the Dapeng Cove,however,reasonable regulation should be made for the sustainable development of the ecosystem function of this cove.

    • Research on the optimal protein to lipid(energy)ratio of blunt snout bream(Megalobrama amblycephala)in two growth stages

      2014, 38(6):861-868. DOI: 10.3724/SP.J.1231.2014.49126

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      Abstract:In order to estimate the appropriate protein to lipid(P/L)ratio of blunt snout bream(Megalobrama amblycephala)for semi-purified diet in different growth stages,2 feeding trials including diets with different P/L levels were conducted in the same formula system.Casein and Peruvian fish meal were used as the source of protein,while soybean oil and soy lecithin were used as lipid source.Dietary protein concentration ranged from 26.93% to 35.65%,and the lipid concentration range was correspondingly between 10.24% and 3.35%.The experimental diets contained 6 gradients,and P/L ratios were 2.63,3.20,4.07,5.33,7.25,10.64,respectively.Test one,two respectively used blunt snout bream with initial average weight of (35.07±0.45) g(juvenile stage),(101.65±1.82) g(premature stage),and were divided into 6 groups,each with 4 replicates.Experiments on the two tests were conducted in pond cages.Both the two tests were fed with the above-mentioned feed,precisely and on schedule.The feeding time were respectively 85 d,and 56 d.The results of 2 experiments showed that:with the feed P/L ratio gradually increased,the evaluation indexes such as the specific growth rate(SGR),the rate of protein deposition(PDR)and energy retention(ERR)increased first and then decreased.The feed conversion ratio(FCR)was the first decline after rising trend.The best growth appeared in the group of P/L 5.33.SGR,FCR,PDR and ERR were used as sensitive criterions for regression analysis,and it showed that different growth stages of blunt snout bream need different optimal P/L ratios:Appropriate P/L ratio of juvenile stage was 6.09-7.58,the optimal protein concentration was 32.83%-33.98% and lipid concentration was 4.48%-5.39%;In the conditions of the average daily intake of 5.06 g/100 g body weight,protein requirement was 1.66-1.72 g/100 g body weight,and lipid requirement was 0.23-0.27 g/100 g body weight for juvenile stage.Appropriate P/L ratio of premature stage was 4.03-4.34,the optimal protein concentration was 30.19%-30.72% and lipid concentration was 7.07%-7.49%.In the conditions of the average daily intake of 3.61 g/100 g body weight,protein requirement was 1.09-1.11 g/100 g body weight,and lipid requirement was 0.26-0.27 g/100 g body weight for premature stage.

    • Comparative analysis of genetic variation in cultured and wild populations of black rockfish(Sebastes schlegelii) based on mitochondrial DNA control region

      2014, 38(6):769-777. DOI: 10.3724/SP.J.1231.2014.49006

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      Abstract:To investigate the genetic variation and diversity of black rockfish Sebastes schlegelii among four cultured populations and six wild populations,the 451bp hypervariable fragments of mitochondrial DNA control region were sequenced and analyzed.The results revealed that the gene diversity indices of cultured populations ranged from 0.540±0.067 to 0.815±0.021,which were significantly lower than those of wild populations(0.883±0.053-0.944±0.028),while the values of nucleotide diversity for both cultured and wild populations were at low level.Among the 52 haplotypes defined,only 12 of them were in the possession of cultured populations which shared 6 haplotypes with the wild populations.The results of pairwise Fst and AMOVA indicated that significant genetic differentiation existed among cultured populations,as well as between wild and cultured populations,which was contrary to that detected among wild populations.No significant genetic differentiations were detected between cultured and wild groups.Significant genealogical structure was not detected either corresponding to the cultured or wild populations based on the MST and NJ analyses.

    • Effect of different cyclic starvation and refeeding regimes on the compensatory growth of Nile tilapia(Oreochromis niloticus)

      2014, 38(6):869-876. DOI: 10.3724/SP.J.1231.2014.49075

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      Abstract:A 43-day feeding trial was conducted to assess the compensatory growth response to different cyclic starvation and refeeding regimes in Nile tilapia(Oreochromis niloticus)fed the diet containing 33% crude protein and 8% lipid.Seven feeding regimes were designed as follows:the control group of fish fed daily(S0,control),and other six groups that were subjected to 1 or 2 days of starvation followed by 3,5 or 7 days of feeding to satiation,i.e.S1F3(starved 1 d,then fed 3 d),S1F5(starved 1 d,then fed 5 d),S1F7(starved 1 d,then fed 7 d),S2F3(starved 2 d,then fed 3 d),S2F5(starved 2 d,then fed 5 d)and S2F7(starved 2 d,then fed 7 d).Four hundred and twenty fish averaging 13.50 g were randomly distributed into twenty one 150-L-tanks(twenty fish per tank)and reared in circular flow-through fresh water.Each of the groups was alloted the diet three times a day for their respective refeeding days.The weight gain rate(806.74%)in group S0 was 40.3%,33.6% and 10.4% higher than those in groups S2F3,S2F5 and S2F7(P<0.05),but was not different from those in groups S1F3,S1F5 and S1F7(P>0.05).The lowest specific growth rate(5.36%)was found in group S0,and was significantly lower than those in other groups(P<0.05).Feed efficiency,protein efficiency ratio,hepatosomatic index,viscerasomatic index,and condition factor as well as whole-body ash content and muscle RNA/DNA ratio did not differ across treatments(P>0.05),but the daily feed intake in groups subjected to any of the cyclic feeding regimes was significantly higher than that in group S0(P<0.05).The groups S2F3,S2F5 and S2F7 exhibited lower whole-body lipid and protein contents than group S0(P<0.05).However,groups S1F3,S1F5 and S1F7 did not show any difference in whole-body composition compared to group S0(P>0.05).In terms of blood biochemical components,total cholesterol,high density lipoprotein cholesterol,low density lipoprotein cholesterol and urea nitrogen contents did not differ across the treatments(P>0.05).The value for triglycerides in group S0 was found the highest,and was higher than those in groups S2F3,S2F5 and S2F7(P<0.05).However,the value for glutamic-pyruvic transaminase was the lowest among treatments,and was lower than those in groups S2F3,S2F5 and S2F7(P<0.05).The serum activity of glutamic-oxalacetic transaminase in the control was similar to those in groups S1F3,S1F5 and S1F7,but lower than those in groups S2F3,S2F5 and S2F7(P<0.05).The results indicated that completely compensatory growth was observed in the tilapia subjected to 1 day of starvation followed by 3,5 or 7 days of feeding to satiation,but partial compensation was found in the fish subjected to 2 days of starvation followed by 3,5 or 7 days of feeding.Hyperphagia was triggered by the cyclic feeding regime,thus feed intake increased,resulting in growth compensation.Based upon the effect of compensation and actual days of feed intake,the cyclic feeding regime of S1F3 could be recommended for the tilapia rearing under the present experimental conditions.

    • Effects of dietary protein and lipid levels on growth of bullfrog(Rana catesbeiana)

      2014, 38(6):877-887. DOI: 10.3724/SP.J.1231.2014.49084

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      Abstract:A feeding trial was conducted to estimate the optimum dietary protein and lipid levels for bullfrog(Rana catesbeiana).Twelve diets were formulated to contain three protein levels(35%,40% and 45%),each with four lipid levels(4%,7%,10% and 13%),in order to produce a range of P/E ratios(from 21.9 to 32.0 mg protein/kJ).The designed dietary protein and lipid levels in a brown fish meal-based formulation were achieved by serial adjustment of casein(for protein)or a mixture of fish oil and soybean oil(for lipid)at the expense of microcrystalline cellulose.Each diet was randomly assigned to three tanks(200 L),each tank was stocked with 12 bullfrogs[(91.5±0.10)g average initial body weight].Bullfrogs were fed twice daily(08:00 and 17:00)to apparent satiation for 8 weeks.The results showed that no significant differences in survival were found among all dietary treatments,but bullfrogs fed the diet with 13% lipid displayed liver lesions,such as abnormal color and crumbly texture.Weight gain(WG)and specific growth rate(SGR)of bullfrogs fed diet with 40% protein were significantly higher than those of bullfrogs fed diet with 35% protein(P<0.05),but there were no significant differences with bullfrogs fed diet with 45% protein(P>0.05).With increasing dietary protein levels,feed efficiency(FE)increased significantly,whereas,feed rate(FR)and protein efficiency ratio(PER)were just the opposite(P<0.05).WG,SGR and FR increased significantly with increasing dietary lipid levels(P<0.05),however,FE,PER and nitrogen retention were not significantly affected by dietary lipid levels(P>0.05).No significant difference was observed in body composition among all the groups at different dietary protein levels(P>0.05).Whole-body crude protein contents were not significantly affected by dietary lipid levels(P>0.05).However,whole-body crude lipid and energy contents increased significantly with increasing dietary lipid level(P<0.05).With increasing dietary lipid levels,hepatic malondialdehyde(MDA)concentration increased significantly,whereas,hepatic catalase(CAT)and lipoprotein lipase(LPL)activity were first increased and then decreased(P<0.05).The results of this study suggest that diet containing 40% protein and 7% lipid with a protein to energy ratio of 27.7 mg protein/kJ is optimal for bullfrog.

    • Comparison of morphology between two kinds of hybrid groupers and their parents

      2014, 38(6):778-785. DOI: 10.3724/SP.J.1231.2014.48939

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      Abstract:In order to find the morphological differences between the two kinds of hybrid groupers and their parents,Hybrid 1(Epinephelus coioides♀ ×Epinephelus lanceolatus♂)and Hybrid 2(Epinephelus fuscogutatus♀ ×Epinephelus lanceolatus♂),we measured their body and analyzed meristic data and metric characters by using the method of statistics,including ANOVA analysis and three types of multivariation analyses.The results showed that the edge of inclined belt on Hybrid1's body was clear and coherent,and there were large annular plaques on Hybrid 2,all those were different from their parents.Furthermore,the analysis based on the morphology framework showed the shape of front dorsal fin and trunk of Hybrid 1was inclined to its female parent,however,Hybrid's front dorsal fin shape was also similar to its famale parent while its trunk shape resembled its male parent,which proved that it was a hybrid of two forms.Comprehensive statistical analysis,such as cluster analysis,discrimination analysis and principal component analysis,indicated that both kinds of hybrids resembled their female parents in morphological characteristics.

    • Cloning and expression analysis of ecdysteriod receptor gene (Ers-EcR)in Eriocheir sinensis

      2014, 38(6):786-793. DOI: 10.3724/SP.J.1231.2014.49039

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      Abstract:Ecdysteroid receptor(EcR)could mediate the regulation effect of ecdysteroid hormone(EH),and plays important roles in molting and growth of Arthropod.To study the structure and function of ecdysteroid receptor(EcR)gene in Eriocheir sinensis,a complete cDNA sequence encoding EcR(Ers-EcR,GenBank Accession No:KF736985)was cloned from Y-organ of E.sinensis by 5' and 3' RACEs.The full-length cDNA consists of 2 176 bp with a 1 638 bp open reading frame,encoding 545 amino acids without signal peptide.Typical domains of nuclear receptor super family:DNA binding domain(DBD)and ligand binding domain(LBD)were identified by SMART(simple modular architecture research tool)in Ers-EcR.DNA binding domain contains 8 conserved cysteines forming two zinc finger motifs(C156-C159-C173-C176,C192-C198-C208-C211),which is the typical character of DNA binding domain.Multi-sequence alignments analysis showed Ers-EcR shared the highest identity with Uca pugilator(91%),also shared high identities with Callinectes sapidus(88%),Scylla paramamosain(87%)and Portunus trituberculatus(83%),however,it showed lower identity with EcRs from shrimps and insects.Phylogenetic analysis revealed that Ers-EcR was closely related to U.pugilator and Gecarcinus lateralis.Tissue distribution of Ers-EcR was detected by quantitative real-time PCR,although strong expression of Ers-EcR was detected in muscles and Y-organ,and its expression could also be found in eyestalk,intestine,hepatopancreas,heart,haemocyte,ovary,testis,thoracic nerve cord and gill.This study would facilitate the study of the roles of Ers-EcR in molt and growth regulation in crustacean.

    • Integrated pharmacokinetics/pharmacodynamics parameters of florfenicol in crucian carp(Carassius auratus)and grass carp(Ctenopharyngodon idella) and their significances in drawing medication regimen

      2014, 38(6):888-895. DOI: 10.3724/SP.J.1231.2014.48983

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      Abstract:This study aims to investigate the differences of in vitro pharmacodynamics(PD)parameters and in vivo pharmacokinetic(PK)of florfenicol in crucian carp(Carassius auratus)and grass carp(Ctenopharyngodon idella)against a pathogenic Aeromonas hydrophila strain(CAAh01),which were used to draw reasonable medication regimen to control disease and prevent drug resistant bacteria.The pharmacodynamics of florfenicol on bacterium CAAh01 were studied in vitro and the pharmacokinetic parameters of florfenicol in crucian carp and grass carp were monitored after a single oral gavage of 10,20,30 mg per kg body weight to determine a medication regimen to prevent bacterial sepsis in grass carp and crucian carp.The in vitro pharmacodynamic results showed that MIC of florfenicol on CAAh01 was 0.5 μg/mL,and MBC was 1.0 μg/mL; the MPC was 6.0 μg/mL(12MIC); the mutant selection window(MSW)of florfenicol on the pathogenic bacterial strains was 0.5-6.0 μg/mL.The time of florfenicol concentration in serum of crucian carp above MPC was 5h at the dosage of 10 mg/kg; 8 h at the dosage of 20 mg/kg,24 h at the dosage of 30 mg/kg,respectively;AUC24/MIC and Cmax/MIC of florfenicol in serum were 177.06 and 15.59 at the dosage of 10 mg/kg,respectively; 265.90 and 21.32 at the dosage of 20 mg/kg,respectively; 426.50 and 31.24 at the dosage of 30 mg/kg,respectively.The time of florfenicol concentration in serum of grass carp above MPC was 0h at the dosage of 10 mg/kg and 20 mg/kg,3h at the dosage of 30 mg/kg,respectively;AUC24/MIC and Cmax/MIC of florfenicol in serum were 38.60 and 4.75 at the dosage of 10 mg/kg,respectively; 75.08 and 10.08 at the dosage of 20 mg/kg,respectively; 121.94 and 19.99 at the dosage of 30 mg/kg,respectively.Considering the drug remained in the plasma with a concentration above MPC and AUC24/MIC≥100 or Cmax/MIC>8,florfenicol is determined to be applicable for the prevention and treatment of bacterial diseases of crucian carp.The recommendation of the dosing regimen to prevent the selection of drug-resistance is 30 mg/kg,at the intervals of 24 h for 1 day,and the proposed withdrawal period should not be less than 20 d.However,it is not suitable to be used continuously to control and treat bacterial diseases in grass garp.

    • Cloning and transcription analysis of genes coding for BCCP subunit of acetyl-CoA carboxylase in Myrmecia incisa

      2014, 38(6):794-802. DOI: 10.3724/SP.J.1231.2014.49124

      Abstract (2354) HTML (0) PDF 2.85 M (1479) Comment (0) Favorites

      Abstract:Acetyl-CoA carboxylase(ACCase)catalyzes the biotin-dependent carboxylation of acetyl-CoA to produce malonyl-CoA,which is a key and rate-limited step in the biosynthesis of fatty acids.In order to investigate its effect on the arachidonic acid(ArA)in Myrmecia incisa grown under nitrogen starvation,two genes coding for biotin carboxyl carrier protein(BCCP)which was one subunit of the heteromeric ACCase complex,were cloned by using Rapid Amplification Complementary DNA ends(RACE)method and reverse transcription(RT)-PCR with designed degenerated primers.The full-length cDNA sequence of MiBCCP1 was composed of 1 267 bp,comprising a 44-bp 5'-untranslated region(UTR)and a 524-bp 3'-UTR with a typical poly A tail.The open reading frame(ORF)was 699 bp in length encoding a 232-amino-acid protein.After restriction of a 45-amino-acid chloroplast signal peptide,the putative molecular weight of mature MiBCCP1 was about 20 ku.The coding sequence of MiBCCP2 gene was also cloned and it was 789 bp in length encoding a 263-amino-acid protein with a 49-amino-acid chloroplast signal peptide,too.The putative molecular weight of mature MiBCCP2 was approximately 22 ku but it lacked a characteristic biotinylation domain at the C-terminus.Neighbor-joining(NJ)dendrogram showed that these two cloned genes belonged to BCCP1 and BCCP2 clades,which was supported by the bootstrip value of 100.Semi-quantitative reverse transcription(RT)-PCR result showed that during the stress course of nitrogen starvation,the relative transcription of both MiBCCP1 and MiBCCP2 genes increased temporally possibly due to shock,and subsequently declined to the end of this experiment,suggesting that M.incisa had a trend to a declined ability of fatty acid biosynthesis de novo.Taking the significantly increased ArA level in this microalga during this culture course together,it was supposed that the homomeric ACCase in the cytosol playsed a more important role than the heteromeric one in modulating biosynthesis and accumulation of ArA in M.incisa under the nitrogen starvation stress.

    • A comparison study on immune function characterization of two shell colours of golden apple snail(Pomacea canaliculata)

      2014, 38(6):896-903. DOI: 10.3724/SP.J.1231.2014.49128

      Abstract (2568) HTML (0) PDF 17.88 M (1556) Comment (0) Favorites

      Abstract:With the aim of exploring immunological difference between black and yellow shell-coloured Pomacea canaliculata,an acute toxicity test of niclosamide was carried out,and their mortality of niclosamide,the cytochemistry characterization and neutral red retention time(NRRT)of hemocytes were investigated in two kind of snails.The results showed the LC50 value of niclosamide for the black-shell snails were 0.77 mg/L and 0.46 mg/L at 24 h and 48 h,and they were higher than the yellow-shell ones.Cytochemical stains(alkaline phosphatase,acid phosphatase,peroxidase,sudan black B,periodic acid schiff,neutral red)showed that most of positive hemocytes were located in the cytoplasm,and the granulocytes were strongly stained.The percentage of positive hemocytes to ACP,AKP,POX of black snails was higher than that in the yellow ones,especially the value of AKP and POX(P<0.01).The intensity of PAS staining in the black snails was stronger than the yellow ones.However,there was no significant difference between NRD and SBB staining.NRRT indicated that the time of neutral red retention in hemocytes of black snails was 9.15 min,which was 1.8 times longer than the yellow ones.This implied that hemocytes of the black snails have higher lysosomal stability.Therefore,the susceptibility of niclosamide,AKP,POX,glycogen and lysosomal stability could affect the immunological function of P.canaliculata.

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