The effect of dietary protein replacement by crystalline amino acid on growth and nitrogen utilization of turbot Scophthalmus maximus juveniles

A study was conducted to evaluate the effect of partial replacement of dietary fish meal by crystalline amino acids on growth performance, feed utilization, body composition and nitrogen utilization of turbot juveniles.

Four diets were formulated to be isolipidic (12% DM) and isonitrogenous (8% DM). A fish meal based diet was used as control. In the experimental diets, a crystalline amino acid (AA) mixture was used to partially replace fish meal, corresponding to a non-protein nitrogen content of 19, 37 and 56%, respectively (diets 19AA, 37AA and 56AA, respectively). The overall amino acid profile of the experimental diets resembled that of the whole-body protein of turbot. Each experimental diet was fed to triplicate groups of 20 fish (initial body weight of 31.8 g) twice daily to apparent satiation for 42 days. During the trial water temperature averaged 18 °C.

Final body weight, weight gain (g kg ABW^− 1 day^− 1) and specific growth rate were not different between the control and 19AA diet but significantly decreased with the increase of crystalline-AA inclusion from 19 to 56%. Feed intake and feed efficiency of fish fed the control and diet 19AA were similar and significantly higher than those of fish fed the 56AA diet. At the end of the growth trial, there were no significant differences in whole-body composition among groups. Hepatosomatic index was also unaffected by dietary treatments.

Nitrogen retention (g kg ABW^− 1 day^− 1) of fish fed the control and the 19AA diets were similar and significantly higher than that of fish fed the other diets. Expressed as a percentage of the nitrogen intake, N retention was significantly higher with the control than with the 37AA and 56AA diets.

Daily ammonia excretion (mg kg ABW^− 1 day^− 1) of fish fed the control diet was significantly higher than that of fish fed the 37AA and 56AA diets, while daily urea excretion (mg kg ABW^− 1 day^− 1) did not significantly differ among treatments. Non-fecal nitrogen (ammonia + urea) excretion (mg kg ABW^− 1 day^− 1) was significantly higher for fish fed the control diet than in those fed the 37AA and 56AA diets. However, as percent of N intake, ammonia excretion and non-fecal N excretion were significantly higher in fish fed the 56AA diet than in those fed the control and 19AA diets.

Specific activity of glutamate dehydrogenase, alanine and aspartate aminotransferases did not significantly differ among experimental groups.

In conclusion, in diets with an overall amino acid profile resembling that of the whole-body protein of turbot, crystalline-AA may replace 19% of dietary protein without negatively affecting growth performances or feed utilization efficiency. However, higher protein replacement levels of protein-bound-AA by crystalline-AA severely depressed growth performance.     

Juvenile growth and survival of heat-shocked triploid hybrid saugeyes, Stizostedion vitreum × S. canadense

Triploid hybrid saugeyes may be a desirable stocking alternative because diploid saugeyes are fertile and can compromise the genetic integrity of parental stocks. Four groups of saugeye eggs were heat-shocked 5 min after fertilization for 15 min at 31 °C, which resulted in an 86.7 ± 9.4% triploidy rate and 57.8 ± 24.3% survival to the eyed-stage. Heat shocked and control saugeyes were stocked in two ponds for 40 days at 395 000 ind. ha^–1. Mean length and weight of triploid and heat-shocked diploid saugeyes were greater ( P  < 0.01) than unshocked diploid saugeyes. Survival of heat-shocked saugeyes (22.4%) was lower than unshocked saugeyes (94.7%). Heat-shocked fish ( n =25 851) harvested from ponds were stocked for 153 days in O'Shaughnessy Reservoir, Ohio, USA. Triploids were 34 mm shorter and weighed 41 g less than heat-shocked diploids ( P  < 0.01). Mean length and weight of triploids were comparable with unshocked diploids elsewhere in Ohio.     

Heat-shock-induced tetraploid and diploid/tetraploid mosaic in pond loach, Misgurnus anguillicaudatus

Tetraploid fish, which are considered as key resources of diploid gametes for further breeding and ploidy manipulation, can be artificially induced by inhibition of the mitotic cell division with hydrostatic pressure or temperature treatments. Although many attempts have been made to induce artificial tetraploid strains, successful establishment of viable and fertile tetraploid strains are rare. In pond loach, Misgurnus anguillicaudatus, natural tetraploid individuals are distributed in wild populations and diploid gametes from the tetraploid fish have been used for the induction of polyploid individuals, but artificially induced tetraploid strains have not been established yet. In the present study, we optimised starting timing of the heat-shock treatment (41 °C for 2 min) to inhibit a mitotic cell division in fertilised eggs of the normal diploid pond loach between 21 and 51 min after insemination at 20 °C. After the treatment, we observed external appearance of hatching larvae and flow cytometrically determined ploidy status of the resultant larvae. Although tetraploid and diploid/tetraploid mosaic larvae were obtained, the optimum timings for induction of tetraploidy varied amongst crosses. Various kinds of ploidy such as haploidy, diploidy, triploidy, pentaploidy, hexaploidy, aneuploidy and mosaic were detected in non-optimum heat-shock timings for tetraploidisation. Survivors, a tetraploid and a diploid/tetraploid mosaic male, matured at the age of 1-year-old, but they produced functional haploid spermatozoa.     

Study of sex determination system in ship sturgeon, Acipenser nudiventris using meiotic gynogenesis

The sex ratios and sex determination mechanism of gynogenetic diploids of ship sturgeon, Acipenser nudiventris, have been investigated to verify the possibilities of sex control by chromosome manipulation in this species. Meiotic gynogenesis was induced in a female ship sturgeon using cold shock after egg activation with UV-irradiated sperm of a male Siberian sturgeon. Microsatellite DNA analysis was applied for verification of uniparental inheritance in the gynogenetic diploid group of fish. All the analyzed gynogenetic diploids possessed only maternal genotype in the examined experimental group of fish. In this study, a minimum of two distinctly selected diagnostic loci in the offspring was used to confirm exclusively maternal contribution. Also, these fish were analyzed for sex diagnostic. Histological analysis of gonads from gynogenetic diploids, obtained from one family, showed 73.3 % of females and 27.7 % of males. The observed sex ratio has suggested that the ship sturgeon have a female heterogametic sex determination system. Gynogenesis in this species with female heterogametic sex determination system will have important role in breeding program and reclamation of its natural population to produce both female and male progeny, while this species has been introduced in the red list of IUCN (International Union for Conservation of Nature and Natural Resources).     

Spontaneous diploidization of the maternal chromosome set in Nile tilapia (Oreochromis niloticus L.) eggs

Spontaneous diploidization of the maternal chromosome set (SDM) in Oreochromis niloticus is described here for the first time. The SDM phenomenon was observed in progeny of only one XY neofemale out of 11 such neofemales from which eggs had been fertilized with UV‐irradiated sperm: this treatment produced only the expected haploid embryos from the other females. SDM progeny were produced from three different batches of eggs from this female. No significant differences in survival at different stages of embryonic development were observed between the SDM and the diploid control group. The maternal inheritance of SDM progeny was verified using multilocus DNA fingerprinting. Only diploid karyotypes were observed in these fish and diploid control groups. In both the SDM and control groups, sex ratios were significantly skewed towards males (93.3% and 65.0% males respectively). The actual mechanism of SDM in eggs from this particular female remains unknown.     

鳜pep和gh基因SNP标记与驯食性状的关联分析

以鳜(Siniperca chuatsi)选育群体为实验材料,在易驯食与不易驯食鳜转录组Unigene数据库中共预测到4809个SNP位点,其中胃蛋白酶基因(pepsinoge, pep)和生长激素基因(growth hormone, gh)均为转录组筛选获得的鳜驯食性状候选基因,本研究将候选基因上的多态SNP位点在易驯食和不易驯食鳜群体中进行基因分型,并与鳜驯食性状进行关联分析。在易驯食与不易驯食的鳜群体中共发现5个单核苷酸(SNP)多态性位点,有效等位基因(N_e)在1.1959～1.7001,观测杂合度(H_o)和期望杂合度(H_e)分别分布于0.1800～0.3585和0.1655～0.4160,多态信息含量(PIC)为0.2477,全部位点都属于中度多态性位点。结果表明SNP位点pep-A T/C中2种基因型TT和CT与鳜驯食性状呈高水平显著相关(P0.05),组合得到的3种基因型Genotype1(CT,CC/CT/TT,AA,AA,TT)、Genotype2(TT,CC/CT/TT, AA, AA, TT)和Genotype3(TT, CC, AA, AA, TT)也与鳜驯食性状呈高水平显著相关(P0.05),显著影响鳜驯食性状表型,其中Genotype2相关性最高,可作为最优基因型个体进行选育。本研究在鳜pep和gh基因中鉴定出与驯食性状呈显著关联的SNP分子标记,为加快易驯食鳜新品种的基因辅助选育提供有效的SNP分子标记。     

Apparent digestibility of protein, energy, and amino acids in some selected feed ingredients for pacu Piaractus mesopotamicus

The apparent digestibility coefficients (ADCs) of amino acids (AA), protein, and energy in six feed ingredients (Brazilian fish meal, soybean meal, corn gluten meal, alcoholic yeast, corn, and wheat bran) we evaluated for pacu juveniles. In general, all ingredients showed high digestibility values for all AA, and differences among ADCs of individual amino acid were detected ( P  < 0.01). Corn gluten, soybean, and fish meals had the highest ADCs of AA. The ADCs of protein in fish meal, yeast, and corn gluten meal should not be used as AA digestibility indicators, because those showed differences up to 6.7% between the ADCs of protein and AA. All ingredients had lower ADCs of energy than corn gluten meal ( P  < 0.01). Lysine was the first limiting amino acid in fish meal, corn gluten meal, wheat bran, and corn, and the second limiting amino acid in soybean meal, as methionine was the first limiting amino acid in soybean meal and yeast. However, the soybean meal was the best quality protein source, as it had the highest digestible essential amino acid index. This demonstrated that digestible amino acid values can be used to formulate practical diets for pacu, preventing potential deficiencies or excess that might cause environmental and economic losses.     

Occurrence of two forms of Y chromosome in rainbow trout (Oncorhynchus mykiss) males from Rutki strain

Morphologically differentiated sex chromosomes are found in only few salmonid species. Some populations of rainbow trout, Oncorhynchus mykiss, exhibit chromosomal polymorphism related to sex. We found sex-related chromosomal polymorphism in fish from the synthetic Rutki strain, Poland, in approximately 85% (n = 22) of examined males (XY-like) whereas the remaining males (n = 4) possessed chromosomes similar to these observed in females (XX-like). To investigate whether males possessing XX-like chromosomes were genotypic males or genotypic females with altered phenotypic sex, androgenetic progeny of four males (representing both XY-like and XX-like forms, n = 2 + 2) was examined. Androgenetic progeny (F₁) of all four fathers consisted of both phenotypic females and males. F₁ male progeny of two fathers showed XX-like chromosomes whereas F₁ male progeny of the other two fathers possessed YY-like (supermale) chromosomes. F₁ were reared further until they were sexually mature. Two males from each of four F₁ families were used to produce F₂ androgenetic and control F₂ generation. All F₂ individuals, androgenetics and control, were phenotypic males at sexual maturation. The results indicate that males possessing XX-like chromosomes are genetic males and they are not sex-reversed females. Thus, the Y chromosome can exist in different morphological forms in farmed rainbow trout. The Y^S chromosome (shorter form, unlike the X chromosome) has a shorter or absent p-arm and lacks the pericentromeric cluster of AT-rich chromatin and the 5S rDNA sequences that are found in the Y^L chromosome (longer form, like the X chromosome).     

Bio‐Ag reutilization of distiller's dried grains with solubles (DDGS) as a substrate for black soldier fly larvae,Hermetia illucens,along with poultry by‐product meal and soybean meal,as total replacement of fish meal in diets for Nile tilapia,Oreochromis niloticus

A feeding trial was conducted in a closed system with Nile tilapia, Oreochromis niloticus, juveniles (mean initial weight, 2.66 g) to examine total replacement of menhaden fish meal (FM) with distiller's dried grains with solubles (DDGS), which had been used as substrate for the production of black soldier fly larvae, Hermetia illucens, in combination with soybean meal (SBM) and poultry by‐product meal (PBM), with or without supplementation of the amino acids (AA) DL‐methionine (Met), L‐lysine (Lys) and a commercial non‐amylaceous polysaccharide enzyme (Enz) product. Fish were fed seven isoenergetic [available energy (AE) = 4.0 kcal g^?1 of diet] and isonitrogenous (350 g kg^?1 protein as‐fed basis) practical diets formulated with equivalent digestible protein levels. Diet 1 was formulated to be similar to a commercial, high‐quality, tilapia diet containing 200 g kg^?1 FM. Diets 2–5 were formulated as a 2 × 2 factorial to replace FM with similar contributions from DDGS (45%), PBM (25%) and SBM (2.1–2.9%), but to differ in supplementation of AA and/or Enz preparation. Diets 6 and 7 were formulated to investigate the effects of a 2/3 and 1/3 reduction, respectively, in DDGS contribution to the replacement protein mix, with concomitant increases in SBM, with respect to diet 3, and were balanced with Lys and Met. After 6 weeks, growth responses were slightly attenuated (P ≤ 0.05) and average daily intake (ADI) and feed conversion ratio (FCR) were slightly higher in tilapia fed DDGS diets 2–5 compared to those of fish fed the FM control diet 1. Growth responses were not significantly affected by the presence or absence of AA or Enz (diets 2–5), or the level of DDGS (diets 3, 7 and 6). Whole‐body proximate composition was not different among treatments. Amino acid profiles of fish fed DDGS diets were not significantly different from those of fish fed the FM control. Evidence of interaction between AA and Enz supplementation was detected in whole‐body amino acid concentrations such that AA content was higher with AA or Enz addition alone, but lower when both were added to the diet. Results suggest that DDGS replacement of FM in tilapia diets can be substantial when diets are formulated on a digestible protein basis and DDGS is combined with highly digestible animal (PBM) and plant proteins (SBM).     

黄姑鱼异质雌核发育子代的遗传分析

为了解黄姑鱼（Nibea albiflora）异质雌核发育子代的基因纯合情况,利用微卫星标记（SSR）和扩增片段长度多态性标记（AFLP）对黄姑鱼异质雌核发育家系进行遗传鉴定和分析。结果显示：（1）雌核发育家系在4个SSR位点和5对AFLP引物组合扩增出的位点均未发现父本特异性等位条带,表明雌核发育体比率为100%。（2）用于遗传分析的7个SSR位点在雌核发育家系和正常交配家系中均未见完全纯合的情况,雌核发育家系7个SSR位点的平均纯合度为0.382,是正常交配家系（0.161）的2.37倍。雌核发育家系各个体的纯合位点数为0~6个,纯合位点所占比例为0~85.7%。（3）5对AFLP引物共扩增出182条清晰的扩增条带,其中有21条父本特异性条带和16条母本特异性条带。16条母本特异性条带中有7个条带在雌核发育家系中显著偏分离（P<0.05）。雌核发育家系和正常交配家系多态性条带比例分别为14.7%和20.3%。（4）雌核发育家系与母本的遗传相似度高于与正常交配家系的遗传相似度,正常交配家系同父本和母本的遗传距离大致相同。研究结果表明,黄姑鱼异质雌核发育二倍体家系的遗传纯合度显著高于正常交配家系,人工诱导雌核发育是促进基因纯合的一个有效途径,它不仅可以加速有利基因的纯合固定,还可以加速有害基因的淘汰,从而有效提高育种效率。     

Survival,growth and fertility of gynogenetic diploids induced in the cyprinid loach, Misgurnus anguillicaudatus

Gynogenetic diploids were induced in the loach, Misgurnus anguillicaudatus, by applying cold shock to the egg after fertilization with spermatozoa genetically inactivated by ultraviolet-ray irradiation, to examine their survival, growth and fertility. Eggs were obtained from orange phenotype loach (homozygous for a recessive demelanogenesis gene). Spermatozoa were obtained from black phenotype loach (homozygous for a dominant melanogenesis gene), and from common carp, Cyprinus carpio. All hybrids between loach and carp were inviably abnormal.Eggs fertilized with loach or carp spermatozoa irradiated at doses of 6000 and 12000 erg/mm ² and then treated with cold water (1°C for one hour 4 and 5 min after fertilization developed into embryos with 61–78% survival. About 70–87% of newly-hatched fry were normal in appearance. Gynogenetic fish induced by both loach and carp spermatozoa showed lower rates of survival and growth than did the normal diploid controls. All gynogenetic fish from carp spermatozoa were female and 94.9% of the gynogenetic fish from loach spermatozoa were female. The second gynogenetic offsprings were produced from females selected from both gynogenetic groups.     

Genetic analysis of aquabirnaviruses isolated from wild fish reveals occurrence of natural reassortment of infectious pancreatic necrosis virus

In this study, we report the sequencing of the whole genome [including the 5' and 3' non-coding regions (NCR) of both segments A and B] of seven birnavirus strains isolated from wild fish from the Flemish Cap (FC) fishery at Newfoundland, Canada. From analysis and comparison of the sequences, most of the FC isolates clustered with the North American reference strains West Buxton (WB), Dry Mill and Jasper. One strain was included in the same genotype as the European strain Ab. In addition, at least in one case cohabitation of both type strains in an individual fish was demonstrated. These results clearly suggest the acquisition of the viruses from two different sources. The prevalence of the American type is easily explained by the close proximity of this fishing bank to the American coast whereas, although surprising, the presence of the European type strain could be because of migration of fish from European waters. In one strain, segment A and B sequences were typed differently (WB and Ab, respectively). These findings indicate natural reassortment between two strains of aquabirnaviruses in a host.     

正常和白化褐牙鲆皮肤和肌肉中DHA和EPA含量的比较

利用气相色谱法对相同养殖条件下生长的正常和白化褐牙鲆(Paralichthys olivaceus)的皮肤和肌肉组织中的DHA和EPA含量进行了比较分析。结果显示，正常和白化褐牙鲆的皮肤和肌肉中EPA含量相近，但白化个体皮肤中DHA含量显著低于正常个体，约为后者的1／2。而两者肌肉中的DHA含量则没有显著差异。同时，白化个体皮肤和肌肉组织中的DHA／EPA比值略低于正常个体。这一结果证明白化褐牙鲆皮肤蓄积或利用DHA的能力要低于正常褐牙鲆。关于鲆鳔类白化现象同鱼体对饵料中脂肪酸类物质的利用率之间的关系有待深入研究。     

Weaning of Atlantic halibut Hippoglossus hippoglossus L. using formulated diets with various levels of ascorbic acid

Atlantic halibut larvae (120 mg) were weaned to formulated diets with different supplementations of ascorbate- poly-phosphate, ApP (300, 2000 and 3000 mg ascorbic acid (AA) equivalents kg⁻¹ diet). The experiment lasted for 50 days with cofeeding of enriched Artemia and formulated diets during the first 30 days. During the last 20 days, only formulated diets were offered to the fish. One control group was fed only Artemia (770 mg AA kg⁻¹ dry weight) during the entire experimental period. The specific growth rate during the 50 days was ≈ 4.5% day⁻¹ and the mean weights in all dietary groups were ≈ 1 g when the experiment was terminated. No differences in mean weight and mortality were observed between the groups fed formulated diets and that fed Artemia during the experiment. The fish size in the groups fed formulated diets ranged between 0.10 and 3.05 g and this differed from the Artemia group where the size ranged between 0.35 and 1.35 g. Dietary levels of ApP had no positive effect on growth and survival. The retention of AA was significantly higher in the groups fed high dietary levels of ApP. Apparently, the bioavailability of high dietary levels of ApP appeared to be low for young halibut. After stressing the fish using a high-salinity challenge test, no significant difference in survival occurred among the dietary groups. Cortisol levels in plasma recorded 3 h post stress was significant lower in the Artemia group compared with the groups fed the formulated diets.     

大黄鱼22个微卫星标记在F1家系中的分离方式及与生长性状的相关分析

研究了1个大黄鱼F1家系150个个体22个微卫星位点的基因型分布,并分析了标记位点与生长性状的相关性。结果显示,22个位点共检测到60个等位基因,平均等位基因数为2.73,平均有效等位基因数为2.37;平均观测杂合度与期望杂合度分别为0.75和0.73,部分位点基因型分布严重偏离孟德尔定律,暗示其可能与适应性基因相连锁,其中LYC0446位点附近可能存在隐性纯合致死基因。LYC0077位点与体质量、体长和体高均呈显著相关(P<0.05),其中等位基因A(165 bp)对应的生长性状表型值最大,可以作为选育快长性状的有效分子标记;LYC0015和LYC0243与体高呈显著相关(P<0.05),与体长、体质量的相关不显著(P>0.05),LYC0015位点的等位基因C(110 bp)和LYC0243位点的等位基因A(160 bp)为有利的等位基因。对LYC0015、LYC0077和LYC0243进行不同基因型个体表型值的多重比较,找到3种对生长性状有利的基因型,分别为BC、AA和AB。同时,以体质量性状为参照,对3个位点不同基因型组合进行比较,找到一个最优基因型组合(BC/AA/AB),与3个位点单独分析对应的最优基因型完全一致,符合加性作用模型。     

Assessing the potential for fish predation to impact zebra mussels (Dreissena polymorpha): insight from bioenergetics models

Abstract –  Rates of annual food consumption and biomass were modeled for several fish species across representative rivers and lakes in eastern North America. Results were combined to assess the relative potential of fish predation to impact zebra mussels ( Dreissena polymorpha ). Predicted annual food consumption by fishes in southern waters was over 100% greater than that in northern systems because of warmer annual water temperatures and presumed increases in metabolic demand. Although generally increasing with latitude, biomasses of several key zebra mussel fish predators did not change significantly across latitudes. Biomasses of some less abundant fish predators did increase significantly with latitude, but increases were not of the magnitude to offset predicted decreases in food consumption. Our results generally support the premise that fishes in rivers and lakes of the southern United States (U.S.) have inherently greater potential to impact zebra mussels by predation. Our simulations may provide a partial explanation of why zebra mussel invasions have not been as rapid and widespread in southern U.S. waters compared to the Great Lakes region.     

Age and growth of alfonsino Beryx splendens from the Kanto District, central Japan, based on growth increments on otoliths

ABSTRACT:   Age and growth of alfonsino Beryx splendens (Lowe) were studied using counts of presumed daily incremental growth rings on the transverse section of otoliths of fish collected from the Kanto District, central Japan. Microstructural growth increments were observed from the core to the outermost margin of the broadest of approximately 50 branches formed on the surface of an otolith. Of 98 otoliths examined, 46 were readable. The number of increments and fish lengths ranged from 448 (218 mm fork length [FL]) to 3701 (411 mm FL). The ages of these fish were estimated to be 1 year, 2 months and 10 years, 2 months, respectively, assuming that an increment was formed daily. The von Bertalanffy growth equation combined for males and females was expressed as follows: L _t  = 542 {1 −  exp  [−0.133( t  + 2.00)]}, where L _t is fork length (mm) at yearly age t . The results of the present study were compared with those of other researchers who had studied alfonsino from central Japan.     

Effects of dietary phosphorus and lipid levels on utilization and excretion of phosphorus and nitrogen by rainbow trout (Oncorhynchus mykiss). 1. Laboratory-scale study

Nutritional strategies to reduce both phosphorus (P) and nitrogen (N) excretion relative to growth of rainbow trout were tested in a 2 × 3 factorial experiment. The two factors were `dietary P level' and `dietary lipid level.' Reduction in dietary P from 14 to 8 g kg^–1 dry diet was achieved by partial substitution of dietary fish meal with a combination of full-fat soyabean meal, corn gluten and spray-dried blood meal. Triplicate tanks of 35 rainbow trout per tank were fed experimental diets for 16 weeks and grew from approximately 40 to 250 g, in 15 °C spring water. All tanks were fed the same percent biomass per day. Diets were isonitrogenous, and dietary energy varied with dietary lipid. Diet digestibility data and results of the experiment were used to construct N and P budgets for the fish fed the various diets. A reduction in dietary fish meal from 500 to 200 g kg^–1 dry diet, corresponding to a reduction in dietary P from 14 to 8 g kg^–1 dry diet, resulted in >50% reductions in both solid and dissolved P waste, but did not affect growth, feed efficiency ratio (FER) or sensory characteristics of rainbow trout. Increasing dietary lipid from 170 to 310 g kg^–1 dry diet led to higher growth rate and FER, and lower total N waste relative to weight gain, but did not change protein retention. Increasing dietary lipid level increased deposition of lipid in whole bodies of rainbow trout, and resulted in discernible differences in sensory characteristics of trout fillets.