Induced meiotic gynogenesis in tench, Tinca tinca (L.) using irradiated heterogenic sperm

Effects of the starting time and duration of cold shock, as well as the source of heterogenic sperm on the percentage of viable gynogenic larvae (PVGL) in tench were studied. The DNA in sperm of red common carp (Cyprinus carpio var singuonensis) was inactivated by ultraviolet radiation prior to use to induce meiotic gynogenetic development in tench. In experiment 1, tench eggs were cold-shocked for 30 min starting at 1, 3, 5, 7, 10 and 15 min post activation. In experiment 2, cold shock began 5 min after activation and lasted for 10, 20, 30, 40, 50 min, respectively. Each experiment was run in triplicate using 3 tench females, and one group not treated with cold shock was included in each experiment to serve as a control group. In experiment 3, sperm of bigmouth buffalo (Ictiobus cyprinellus), red common carp or grass carp (Ctenopharyngodon idella) was used to induce gynogenesis in tench with a 20-min cold shock starting 5 min post activation. The results showed that PVGL from the control group was very low (0.11–0.47%). In experiment 1, the highest average PVGL (9.60%) was observed when cold shock treatment was applied 5 min post activation. When cold shock treatment was started 5 min post activation, duration of cold shock affected PVGL. Cold shock lasting 20 min resulted in the highest average PVGL (12. 57%) among the selected duration of cold shock studied in experiment 2. The average PVGL was 2.3, 8.6, and 9.3%, respectively, for eggs induced by sperm of bigmouth buffalo, red common carp and grass carp. Average PVGL was significantly lower for eggs induced by sperm of bigmouth buffalo, compared with that for eggs induced by sperm of the other two species. However, average PVGL were similar for eggs induced by sperm of red common carp and grass carp. In summary, the optimal conditions for gynogenesis in tench include the use of irradiated sperm of grass carp to activate the eggs and cold shock of 20 min starting 5 min post activation. Since female tench grow much faster to a larger size than male tench, gynogenesis of tench holds great potential for production enhancement.     

Diet and fish choice of Eurasian otters (<Emphasis Type="Italic">Lutra lutra</Emphasis> L.) in fish wintering ponds in Hungary

The diet composition and fish preference of piscivorous Eurasian otters (Lutra lutra) were studied in two fish farm systems in Hungary using spraint (otter faeces) analysis during two wintering periods. The primary food source of otters in both fish farms was fish (97–99% of biomass). The main fish prey was small-sized, below 100 g in weight (96% in both areas), while fish prey above 500 g comprised only 0.1–0.4% of the diet. The bulk of the otters’ diet consisted of less-valued species, especially non-native Prussian carp (Carassius auratus gibelio). Consumption of commercial fish species ranged between 15 and 31% of the total diet. Otters preferred fish below 100 g in weight (Ivlev’s electivity index, E _i = 0.65–0.70), and showed a lesser preference for (or avoided) fish above 100 g in weight (E _i = −0.37–1.00). With regard to species distribution, otters preferred small (below 100 g) grass carp (Ctenopharyngodon idella), zander (Sander lucioperca), pike (Esox lucius), Prussian carp, topmouth gudgeon (Pseudorasbora parva), while they consumed common carp (Cyprinus carpio), the most important commercial species, proportionally to its abundance in the environment (E _i = −0.18–0.29).     

Studies on triploid oysters in Australia: Evaluation of cytochalasin B and 6-dimethylaminopurine for triploidy induction in Sydney rock oysters Saccostrea commercialis (Iredale and Roughley)

Naturally spawned Sydney rock oysters Saccostrea commercialis (Iredale and Roughley),were used to determine the appropriate stage of development for inducing triploidy and to compare the effectiveness of cytochalasin B (CB) and 6-dimethylaminopurine (6-DMAP) in dose-optimization trials. Induction should commence at 50% first polar body (PB1) extrusion in eggs (approximately 17-19 min post-fertilization at 25^oC). By day 5 the highest triploidy percentage and yield (number of triploid larvae per 100 fertilized eggs) were achieved in the ranges of 0.75-1.5 mg CB 1^-1 (1.6-3.1 μm CB)or 200-400 μm 6-DMAP (32.6-65.3 mg 6-DMAP l^-1). However, CB treatment resulted in greater survival and triploidy percentage than 6-DMAP in Sydney rock oysters.     

基于微卫星评估草鱼放流亲本对野生群体遗传多样性的影响

利用筛选的13对草鱼多态性微卫星标记,开展了2011至2015年长江中游草鱼亲本增殖放流对野生群体遗传多样性的影响评估。通过对各位点的遗传多样性分析,13个微卫星位点的多态信息含量为0.8622(0.657~0.950),基因多样度为0.8555(0.675~0.936)。15个群体的有效等位基因数为7.4503~10.1536,等位基因丰度为11.483~15.204,说明15个草鱼群体的遗传多样性水平总体较高。遗传分化指数分析表明,群体间不存在显著遗传分化(FST5%)。通过贝叶斯聚类分析和主成分分析可将草鱼群体分为4个组群,根据分组结果以及来源划分分别对草鱼群体进行AMOVA分析,发现遗传变异大部分来自于群体内个体间,组间及组内群体间的分化水平较低(FCT5%,FSC5%),与FST分析结果一致。研究表明,当前草鱼亲本增殖放流模式对野生群体遗传结构影响不明显。     

Effects of ultra-violet irradiation on sperm motility and diploid gynogenesis induction in large yellow croaker (<Emphasis Type="Italic">Pseudosciaena crocea</Emphasis>) undergoing cold shock

Large yellow croaker, Pseudosciaena crocea, exhibit sexually dimorphic growth, with females growing faster and reaching larger adult sizes than males. Thus, development of techniques for preferentially producing females is necessary to optimize production of these species. We have established a protocol to produce all-female croaker P. crocea through induction of meiotic gynogenesis with homologous sperm. The first set of experiments investigated the ultra-violet (UV) irradiation on sperm motility and duration of sperm activity to determine the optimal UV dosage for genetic inactivation of sperm, yet retaining adequate motility for activation of eggs. Milt from several males was diluted 1:100 with Ringer’s solution and UV irradiated with doses ranging from 0–150 J cm⁻². The results indicated that motility and duration of activity generally decreased with increased UV doses. At UV doses greater than 105 J cm⁻², after fertilization, motility was <10% and fertilization rates were significantly lower. Highest hatching rate was obtained at 75 J cm⁻². A second set of experiments was carried out to determine appropriate conditions of cold shock for retention of the 2nd polar body in P. crocea eggs after fertilization with UV-inactivated sperm by altering the timing, temperature and duration of shock. At 20°C, shock applied at 3 min after fertilization resulted in higher survival rate of larvae at 6 h after hatching. Results of different combinations of three shock temperatures (2°C, 3°C or 4°C) and five shock durations (4 min, 8 min, 12 min, 16 min or 20 min) at 3 min after fertilization demonstrated that shocks of 12 min gave highest production of diploid gynogens. Statistical analysis revealed that maximum production of diploid gynogens (44.55 ± 2.99%) were obtained at 3°C. The results of this study indicate that the use of UV-irradiated homologous sperm for activation of P. crocea eggs and cold shock for polar body retention is an effective method for producing gynogenetic offspring.     

High water-temperature tolerance in photosynthetic activity of <Emphasis Type="Italic">Zostera marina</Emphasis> seedlings from Ise Bay,Mie Prefecture,central Japan

Photosynthetic activities of seedlings of Zostera marina were successively measured using a gas volumeter for 6 days at seven light (0–400 μmol photons/m² per s) and 11 water temperature conditions (5–35°C). The seedlings were collected from mature plants (Ise Bay, central Japan), and stored and cultured in incubators accurately controlled at each test temperature. The maximum gross photosynthesis (P _maxg) was recorded at an optimal water temperature of 29°C after 0 days. After 6 days, P _maxg appeared at 25°C and most plants cultured at 29–30°C bleached and withered after the drastic increase of light compensation point (I _c). On the contrary, at 5–28°C, the photosynthetic activities either changed little (5–25°C) or recovered after a temporal reduction (26–28°C); seedlings survived and looked healthy after being cultured for 6 days. The recovery was thought to be an acclimation to tolerate higher water temperature. As a result, the critical upper water temperature for Z. marina seedlings was proposed as 28°C. The temperature was consistent with the previously reported maximum water temperature in habitats around the southern boundary of Z. marina in the northern hemisphere.     

低鱼粉饲料中添加牛磺酸对青鱼幼鱼生长、肠道修复及抗急性拥挤胁迫的影响

为研究低鱼粉饲料中添加牛磺酸对青鱼幼鱼[初始质量(5.90±0.03)g]生长、肠道修复及抗急性拥挤胁迫的影响,实验以青鱼正常鱼粉组(20%)为对照组,10%鱼粉组为负对照组(I_0组),在低鱼粉饲料中分别添加0.05%(I_(0.05)组)、0.1%(I_(0.1)组)、0.2%(I_(0.2)组)、0.4%(I_(0.4)组)牛磺酸,配制6种等氮等脂饲料,饲养8周。饲养实验结束后,参考生长结果,选择对照组、I_0组、I_(0.1)组、I_(0.4)组进行急性拥挤胁迫实验。结果显示:①与对照组相比,I_0组增重率显著下降,随着低鱼粉饲料中牛磺酸水平的升高,青鱼幼鱼增重率呈先上升后下降趋势,当牛磺酸添加量为0.1%时与对照组差异不显著;②相对于对照组,I_0组的绒毛高度降低、隐窝深度增加,杯状细胞数降低,添加牛磺酸使绒毛高度升高、隐窝深度降低,杯状细胞数和淋巴细胞数增多;③急性拥挤胁迫使青鱼幼鱼血清史质醇、血糖、血清溶菌酶(LSZ)、补体C3、超氧物歧化酶(SOD)和谷胱甘肽(GSH)含量均呈先升高后下降的趋势,其最大值出现在胁迫2 h或者8 h。在整个胁迫期间,I_0组史质醇和血糖含量高于其他各饲料组,I_0组溶菌酶、补体C3、SOD和GSH均低于其他各饲料组。研究表明,在低鱼粉饲料中添加牛磺酸可提高青鱼生长性能、改善肠道结构、增强青鱼的抗急性拥挤胁迫的能力。     

Embryonic development and morphology of eggs and newly hatched larvae of Pacific herring <Emphasis Type="Italic">Clupea pallasii</Emphasis>

The embryonic development and morphology of eggs and newly hatched larvae of the Pacific herring Clupea pallasii were described using laboratory-reared specimens originating from the Miyako Bay stock. The eggs were almost spherical in shape, 1.33–1.46 mm (mean: 1.38 mm) in diameter, and had a thick adherent chorion. They had a segmented pale yellow yolk, no oil globule, and a relatively wide perivitelline space. A subgerminal cavity was observed during the gastrula period, whereas the blastocoel did not appear. Mass hatching occurred by 271 h 45 min after fertilization, and the newly hatched larvae were 7.1–7.7 mm (mean: 7.5 mm) in total length with 53–56 myomeres at 9.6°C. The embryonic development of Pacific herring was substantially similar to that of zebrafish Danio rerio, American shad Alosa sapidissima, as well as Atlantic herring Clupea harengus, and generally followed the basic developmental pattern of teleosts. However, Pacific herring larvae hatched at a more developed stage than some other clupeoids, such as Japanese sardine Sardinops melanostictus, and the progressed developmental stage at hatching could be interpreted as an advanced adaptation.     

Artificial gynogenesis and assessment of homozygosity in meiotic gynogens of spotted halibut (<Emphasis Type="Italic">Verasper variegatus</Emphasis>)

In the present study, methodology of gynogenetic induction in spotted halibut were developed and optimized; the sex ratio of putative meiotic gynogenetic diploids was determined using AFLP-based molecular sexing technique; the homozygosity of gynogenetic population was assessed as opposed to cultivated population. The results showed that high percentage of meiotic gynogenetic diploids were generated when the eggs fertilized with irradiated heterologous sea perch frozen sperm (30–50 mJ cm⁻²) were cold shocked in sea water of −1°C for 40–75 min at 5 min after fertilization. About 15,200 diploid gynogenetic larvae were achieved and they exhibited normal morphology similar to diploid control. The gynogenetic diploids were 100% female, which first confirmed the female homogamete (XX/XY) sex determination in spotted halibut. The genetic analysis showed that the average H _O was, respectively, 0.404 and 0.724 in gynogenetic population and cultivated population, indicating an increase of homozygosity in gynogenetic population.     

The effects of dietary lipid sources and lecithin on the production of giant freshwater prawn <Emphasis Type="Italic">Macrobrachium rosenbergii</Emphasis> larvae in the Mekong Delta region of Vietnam

The giant freshwater prawn, Macrobrachium rosenbergii, is cultured widely in the Mekong Delta region of Vietnam but it is often difficult or expensive for hatchery operators to purchase commercial diets used as a feeding supplement to Artemia nauplii. Therefore, in the present study, the effects of lipid sources and lecithin on the growth and survival rate of M. rosenbergii larvae were examined in order to develop suitable hand-prepared larval diets for seed production of M. rosenbergii in this area. Six egg custard diets consisting of various ratios of lipid (originating from soybean oil and squid oil) and lecithin were used for rearing Macrobrachium rosenbergii larvae. Treatments in which larvae were fed diets containing squid oil exhibited the highest body length and survival rates (7.14–7.43 mm and 51.1–68.1%, respectively), and differed significantly from other treatments (P<0.05). Use of dietary soybean oil yielded the lowest body length and survival rates (6.29–6.75 mm and 22.0–48.7%), respectively). The supplementation of dietary lecithin did not increase final body weight but did improve larval survival rates. The n-3 HUFA content of prawns fed dietary squid oil was higher than those of animals provided with other diets. These results indicated that the most appropriate diet for rearing M. rosenbergii larvae is the diet containing 3% squid oil and 1.5% lecithin.     

Influence of the antibacterial herbs, Solanum trilobatum, Andrographis paniculata and Psoralea corylifolia on the survival, growth and bacterial load of Penaeus monodon post larvae

The indiscriminate use of antibiotics and chemicals in shrimp hatcheries has led to biomagnification and that in turn could lead to rejection of a whole consignment. The application of the bioencapsulation technique as a tool for curative treatment in shrimp larvae was investigated. Herbs having antibacterial properties such as Solanum trilobatum, Andrographis paniculata and Psoralea corylifolia (methanolic extracts) were bioencapsulated in Artemia and fed to Penaeus monodon post larvae PL 1–25. The post larvae were reared in a medium inoculated with pathogenic bacteria such as Pseudomonas aeruginosa, Staphylococcus aureus, Salmonella typhi and Vibrio sp. Post larvae reared in the non-inoculated water and fed with non-enriched Artemia exhibited 90% survival, highest specific growth rate (12.43%) and reduced bacterial load. P. monodon reared in the bacterial inoculated water and fed with the non-enriched Artemia exhibited the lowest survival (10–30%), specific growth rate (8.42–9.1%) and increased bacterial load (2.86 × 10³ to 3.76 × 10⁵ cfu/g). The methanolic extracts of the herbs helped to increase survival and specific growth rate and reduced bacterial load in the P. monodon culture system. Among the three herbal extracts, P. corylifolia enriched Artemia fed post larvae showed the tendency to higher survival (>50%), growth rate (11.5 averaged) and low bacterial load (1.12 × 10⁵ cfu/g). This revised version was published online in August 2006 with corrections to the Cover Date.     

Acute Toxicity of Synthetic Pyrethroid Cypermethrin on the Common Carp (<Emphasis Type="Italic">Cyprinus carpio</Emphasis> L.) Embryos and Larvae

In this study, the toxic effects on the embryos and larvae of the common carp were used as a model to investigate the synthetic pyrethroid pesticide, cypermethrin, which contaminates aquatic ecosystems. Data obtained from the cypermethrin acute toxicity tests were evaluated using the Probit Analysis Statistical Method. The control and eight test experiments were repeated five times. The number of dead embryos significantly increased in response to cypermethrin concentrations 0.0001, 0.001, 0.01, 0.1, 1, 2, 4 and 8 μg l⁻¹ (p<0.05 for each case). The 48 h LC₅₀ value (with 95% confidence limits) of cypermethrin for common carp embryos was estimated at 0.909 (0.256–5.074) μg l⁻¹. Dose–response decreases in hatching success were recorded as 87.4, 85.0, 80.2, 71.4, 56.3, 48.6, 38.8 and 23.5%, respectively. The lowest concentration of cypermethrin (0.0001 μg l⁻¹) produced a significant increase in the number of dead larvae compared to the control group (p<0.05). The number of dead larvae significantly increased with increasing cypermethrin concentrations exposed for 1–96 h (p<0.05). The highest concentration (8 μg l⁻¹) showed the highest larvae mortality. The 96 h LC₅₀ value (with 95% confidence limits) of cypermethrin for common carp larvae was estimated at 0.809 (0.530–1.308) μg l⁻¹. The results of the study suggest that low levels of cypermethrin in the aquatic environment may have a significant effect on the reproduction and development of carp.     

Insemination, fertilization and gamete management in tench, Tinca tinca (L.)

Various procedures for artificial insemination in tench, Tinca tinca (L.) were re-examined with evaluation of fecundity of males and females among different tench strains. The objectives of this study were to enhance fertilization and hatching rates through optimization of the activation solution, the insemination process, the activation of gametes, and the elimination of eggs stickiness. Sperm for all experiments was collected directly into immobilization solution of modified Kurokura solution containing 180 mM of NaCl and stored at 2 °C for 2.5–5 h prior to the experiment. When dechlorinated tap water was used for activation a gamete ratio of 1150 spermatozoa per egg showed the best significant fertilisation and hatching rates. Optimal ratio between eggs (weight in g) and activation solution (in cm³) was 1:1. Different concentrations of activation solutions such as NaCl from 0 to 68 mM (0–136 mOsmol kg⁻¹) without buffer statistically decreased fertilization and hatching rates. The activation solution containing 17 mM of NaCl, 10 mM Tris–HCl, pH 8 and 9 significantly increased fertilization and hatching rates compared to dechlorinated tap water of pH 7 or activation solution containing 17 mM of NaCl, 10 mM Tris–HCl, pH 6 and 7. Adhesiveness of the eggs was successfully removed by incubation in Alcalase and activity: 3.16 Anson units per cm³.     

Purification and characterization of cysteine proteinase inhibitors from crucian carp <Emphasis Type="Italic">Carassius auratus</Emphasis> eggs

Two cystatins (cst-I and cst-II) were purified from crucian carp eggs by acidification and subsequent ion exchange and molecular sieve chromatography. The molecular masses of cst-I and cst-II analyzed by sodium dodecyl sulfate–polyacrylamide gel electrophoresis were 11.9 and 14.4 kDa, respectively, under reducing conditions and 13.5 and 12.7 kDa, respectively, under non-reducing conditions. The cst-I and cst-II molecules were stable after 30 min of incubation at 60 and 50°C, respectively. There was no significant loss in the inhibitory activity of either cst in the pH range 4–11. These two cystatins were able to affect the proteolysis of papain, cathepsin L, and bromelain, but they were unable to inhibit cathepsin B and trypsin. The partial N-terminal amino acid sequences of both cst inhibitors were homologous and that of cst-I was recognized as NH₂-AGIPGGLVDADINDADVQ. This latter fragment shared 88.9% identity to common carp cystatin and 44.4–55.6% to cystatins of other aquatic animals. Based on these results, we conclude that the two cst inhibitors are members of family II cystatin.     

In vitro storage of fish oocytes: effect of storage temperature,media conditions and storage duration on fertilization and larval hatchability of Indian major carp,rohu (Labeo rohita)

The viability of matured oocytes stored in vitro were assessed using carp ovarian fluid (OF) and artificial carp ovarian fluid (ACOF) under different temperature regimes (4, 24, 26, 28 and 30°C) for different storage durations (0, 60, 120, 150 and 180 min). Significantly higher fertilization (74%) was achieved when the oocytes were stored using ACOF and 65% in OF after 180 min at 28°C. Similarly the hatching rates of larvae were significantly higher in the ACOF and OF, that is, 64% and 47%, respectively, after 180 min of storage. The oocytes kept in the storage containers with ACOF having 65% moisture level showed a significantly higher fertilization rate than the 59% moisture level. This study demonstrated that unfertilized matured oocytes (eggs) of rohu can be stored in vitro for 180 min without compromising the viability (fertilization and hatching) to a great extent in OF and ACOF.     

Gynogenesis and sex determination in large-scale loach Paramisgurnus dabryanus (Sauvage)

Gynogenesis was induced using heterologous sperms in large-scale loach, Paramisgurnus dabryanus (Sauvage), in which a ZW/ZZ sex determination was previously proposed. Three microsatellite loci were used to monitor exclusive maternal inheritance of gynogenetic progenies. The results showed that high percentages of meiogynogens were produced at 4 min post-fertilization and mitogynogens were produced at 18 min post-fertilization by heat shocks, while meiotic gynogenesis was induced by cold shocks within a wide period and high heterozygosity was even observed in gynogens produced at 24 min post-fertilization. The sex ratios of the F₁ progenies in three gynogenetic families were significantly deviated from 1:1 expectation with a female bias in two families and a male bias in one family (P < 0.05), and the other four gynogenetic families showed approximate 1:1 sex ratios. Moreover, the self-mating between gynogenetic F₁ progenies and mating between gynogenetic F₁ progenies and normal individuals produced all-female progenies or identical proportions of females and males. The data of sex ratios generally confirmed that the sex determination in large-scale loach was determined by the putative ZW/ZZ system, and the possible reasons causing the biased sex ratios are discussed.     

Toxicity of Diazinon 60 EC for Cyprinus carpio and Poecilia reticulata

Diazinon 60 EC (chemical insecticide, organophosphate, active substance diazinon at a concentration of 600 g l⁻¹) is used in fish-farming in well-founded cases as a biocide to suppress excessive propagation of coarse Daphnia zooplankton. At the concentration of 10 μg l⁻¹ (i.e., 100 g per 1 ha at a mean depth of the pond of 1 m), Diazinon 60 EC can highly selectively eliminate Daphnia zooplankton, causes no harm to fish, and relatively quickly decomposes in the aquatic environment. This has been corroborated by results of acute toxicity tests in fish: the 96 h LC50 is 3 mg l⁻¹ for the guppy (Poecilia reticulata) and 10–25 mg l⁻¹ for the common carp (Cyprinus carpio). As demonstrated by embryonic and larval tests of toxicity for the common carp (C. carpio), Diazinon 60 EC at the concentration of 10 μg l⁻¹ causes no harm to the tested, individuals even in these sensitive (critical) stages of ontogeny.     

Changes in mitochondrial fatty acid composition following temperature acclimation of carp and their possible effects on FoF1-ATPase activity

Carp undergo temperature acclimation of respiratory function by altering mitochondrial ATP synthase (F_oF₁-ATPase) both quantitatively and qualitatively (Itoi et al. 2003). To address such acclimation temperature-dependent changes of F_oF₁-ATPase activity, we investigated in this study the correlation between the fatty acid composition and F_oF₁-ATPase activity in fast muscle of thermally acclimated carp. The quantities of saturated fatty acids of mitochondria from carp acclimated to 10 °C were significantly lower than those of carp acclimated to 30 °C. While mono- and poly-unsaturated fatty acids tended to increase with cold acclimation of carp, the molar concentration of 16:0 aldehyde in mitochondria from the 10 °C-acclimated carp were less than those from the 30 °C-acclimated fish. The specific activities of F_oF₁-ATPase in the 10 °C- and 30 °C-acclimated fish mitochondria were calculated to be 167±22 and 56±10 nmol/min ⋅ mg mitochondrial protein, respectively, the difference being significant at P<0.005. Taken together, the increase in F_oF₁-ATPase activity in fast muscle mitochondria of carp after cold temperature acclimation may be closely related to the increase of unsaturated fatty acids in mitochondria. Abbreviations: BSA - bovine serum albumin; DHA - docosahexaenoic acid; EGTA - ethyleneglycol bis(2-aminoethylether)tetraacetic acid; EPA - eicosapentaenoic acid; F_oF₁-ATPase - mitochondrial ATP synthase; α-F₁-ATPase - F_oF₁-ATPase α-subunit; β-F₁-ATPase - F_oF₁-ATPase β-subunit; HEPES - 2-[4-(2-hydroxyethyl)-1-piperazinyl]ethanesulfonic acid; SDS - sodium dodecyl sulfate; SDS-PAGE - SDS-polyacrylamide gel electrophoresis. This revised version was published online in July 2006 with corrections to the Cover Date.     

Influence of stocking density and type of feed on the rearing of crucian carp, <Emphasis Type="Italic">Carassius carassius</Emphasis> (L.), larvae under controlled conditions

Two experiments were conducted into the rearing of crucian carp larvae under controlled conditions in an experimental closed water system. In both cases, the rearing lasted 21 days. The first experiment concerned the initial stocking density (from 50 to 600 individuals per l), whilst the second one studied the first food offered (two types of Artemia nauplii, decapsulated Artemia cysts and three types of commercial feeds) which were applied at the moment of exogenous feeding commencement. The best results were obtained using 50 larvae per l, whilst there were no significant differences within the range 200–600 larvae per l. The application of dry feed had a negative influence on the survival and other parameters. The highest survival rate was found in the groups fed with freshly hatched Artemia naupli. The results obtained indicate that the rearing of crucian carp larvae may be successfully conducted in very high stocking densities. However, the very high sensitivity of crucian carp larvae to the type of food offered during the initial days of life should be taken into account. The data presented in this article could be very useful in crucian carp larviculture.     

Effect of prebiotic xylooligosaccharides on growth performances and digestive enzyme activities of allogynogenetic crucian carp (<Emphasis Type="Italic">Carassius auratus gibelio</Emphasis>)

The effect of prebiotic xylooligosaccharides (XOS) on the growth performance and digestive enzyme activities of the allogynogenetic crucian carp, Carassius auratus gibelio, was investigated. XOS was added to fish basal semi-purified diets at three concentrations by dry feed weight: diet 1, 50 mg kg⁻¹; diet 2, 100 mg kg⁻¹; diet 3, 200 mg kg⁻¹, respectively. Twelve aquaria (n = 20) with three replicates for each treatment group (diets 1–3) and control treated without XOS were used. Weights of all collected carp from each aquarium were determined at the initial phase and at the end of the experiment, and the carp survival was also determined by counting the individuals in each aquarium. After 45 days, there were significant differences (P < 0.05) in the relative gain rate (RGR), and daily weight gain (DWG) of diets 1–3 were compared with the control. However, the survival rate was not affected (P > 0.05) by the dietary treatments. For enzymatic analysis, dissection produced a crude mixture of intestine and hepatopancreas of each segment to measure. The protease activity in the intestine and hepatopancreas content of fish in diet 2 (487.37 ± 20.58 U g⁻¹ and 20.52 ± 1.93 U g⁻¹) were significantly different (P < 0.05) from that in the control (428.13 ± 23.26 U g⁻¹ and 12.81 ± 1.52 U g⁻¹) and diet 3 (428.00 ± 23.78 U g⁻¹ and 14.04 ± 1.59 U g⁻¹). Amylase activity in the intestine was significantly higher for diet 2 compared to diet 1 and the control. As for amylase in the hepatopancreas, assays showed higher activity in diet 2 (P < 0.05) compared to the rest.