Short-term dietary supplementation with the microalga Parietochloris incisa enhances stress resistance in guppies Poecilia reticulata

Two trials were conducted to determine the effects of dietary enrichments with the microalga Parietochloris incisa , rich in arachidonic acid (ARA), on stress resistance in guppies Poecilia reticulata . The microalga was added to commercial diets as a neutral lipid (NL) extract and its fractions or as broken cells. Experimental diets were applied for a period of 14 days. In trial 1, commercial diets were supplemented with NL (containing 25 mg ARA and 0.11 mg β-carotene g⁻¹ feed), its triacylglycerol (TAG) fraction (containing 25 mg ARA g⁻¹ feed and no β-carotene) and the β-carotene fraction (containing 0.11 mg carotenoid g⁻¹ feed and minute amounts of ARA). Neutral lipid-fed fish demonstrated the highest resistance ( P <0.05) to osmotic stress (32-ppt NaCl), followed by fish fed with diets supplemented with TAG and β-carotene alone, which were more resistant than control ( P <0.05). In trial 2, fish fed diets supplemented with higher levels of broken alga (26.1 mg ARA g⁻¹ feed) were more resistant ( P <0.05) to stress as compared with fish fed lower ARA (16.3 mg g g⁻¹) or an unsupplemented control diet. We suggest a dietary supplementation with broken P. incisa cells to enhance stress resistance in guppies before a stressful event.     

Controlled infection of Poecilia reticulata Peters (guppy) with Tetrahymena by immersion and intraperitoneal injection

Tetrahymena is a protozoan parasite, which infects guppy, Poecilia reticulata Peters, and causes substantial economical losses in commercial farms worldwide. Studies of guppy infected by Tetrahymena require standardized infection protocols. The LD50 for Tetrahymena infection of guppies by intraperitoneal (IP) injection was calibrated, and the level obtained was 946 parasites per fish. Guppy infection with Tetrahymena by immersion, imitating the natural route of infection via the integument, was studied under normal or stress conditions. Exposure to cold and netting (CNI) and to cold only (CI) followed by immersion exposure to 10 000 Tetrahymena per mL resulted in 22.5% and 19.2% mortality, respectively, as compared to 14.2% and 10% in groups that were netted only (NI) or non‐stressed (I). Histopathology revealed that immersion infection resulted in a systemic infection. Lysozyme levels, measured 3 weeks after infection, were significantly higher in the CNI group (288 μg per mg protein) compared with CI‐, NI‐ and I‐treated groups (94.5, 64 and 62.3 μg mg^?1, respectively). There was no evident parasite immobilization activity in body homogenates, suggesting no development of acquired immunity. Re‐infection by IP injection revealed no increase in protection in any of the treatment groups, mortality range of 56.3–75%, higher than in the non‐exposed control (40.6% mortality).     

Using attraction to light to decrease cannibalism and increase fry production in guppy (Poecilia reticulata Peters) hatcheries. II: the effects of light and cannibalistic adults

Predation of newborn fish by cannibalistic adults is a major cause of fry loss in guppy breeding tanks. The efficacy of using light to attract newborn guppies into a safe area, thereby reducing the cannibalism rate, was evaluated in aquaria. The aquaria were divided using small‐mesh netting into a section with adult fish and a safe section without adults, above which a light source was installed to attract the newborns. Both, the presence of cannibalistic adults and the light, increased the number of newborns in the safe section. The effect of the cannibals was pronounced during the day, and the effect of light seemed to be more pronounced at night. The presence of light significantly decreased cannibalism rate after 30 min, but not 16 h after the introduction of newborns. When tested with cannibalistic adults that have not been fed for 5 days prior to testing, cannibalism rate was higher and the effect of light on reducing cannibalism was evident both 30 min and 16 h after the introduction of newborns. The results demonstrated the potential efficacy of using white light in reducing cannibalism and increasing fry yields in guppy hatcheries.     

Improvement of reproductive indices,lysozyme activity,and disease resistance in live‐bearing ornamental fish,Poecilia latipinna using Artemia supplementation with treated yeast cell,Saccharomyces cerevisiae

In the present study, the effects of Artemia supplemented with 2‐β‐mercapto‐ethanol (β‐ME) treated yeast cell, Saccharomyces cerevisiae, on growth and reproductive performance, lysozyme activity, and disease resistance to Aeromonas hydrophila of freshwater ornamental species, Poecilia latipinna, were investigated. Within 60 days, molly fish were fed with three treatments including commercial food (T1), un‐supplemented Artemia (T2), and Artemia supplemented with β‐ME‐treated yeast cell (at concentration of 4 × 10⁷ CFU/L of water) (T3). After the feeding period, the fish were exposed to 100 μl of a suspension (1.1 × 10⁷ cells/ml) of A. hydrophila (BCCM5/LMG3279) and the cumulative mortality rates were recorded for 12 days. No significant difference was found between survival rate and growth performance of P. latipinna except for weight gain that was higher in fish fed through Artemia supplemented with β‐ME‐treated yeast cell compared to control group. Fecundity rate was significantly improved in fish fed using T3 with the maximum amount of 49.5 ± 2.29 per female (p < 0.05). Besides, lysozyme activity was significantly increased in group 3 (p < 0.05). Moreover, lowest fish mortality was significantly observed in this treatment (p < 0.05). In addition, the number of colonies formed by yeast cell in T3 (634 × 10³ CFU/g intestine) showed significant difference with other treatments (p < 0.05). In sum, Artemia enriched with β‐ME‐treated yeast improved reproductive indices, immune responses, and resistance against A. hydrophila of P. latipinna.     

Using attraction to light to decrease cannibalism and increase fry production in guppy (Poecilia reticulata Peters) hatcheries. I: phototactic reaction and light colour preference

A substantial proportion of newborns are lost in guppy hatcheries to cannibalism by the broodfish although small‐mesh baskets, which allow passage to small fry only and provide a predation‐free area, are used in breeding tanks. If newborn guppies are attracted by light, illuminating the small‐mesh baskets might enhance aggregation of newborns in the mesh basket and thus mitigate cannibalism effect. In this study, light attraction and light colour preference in newborn guppies were investigated. Testing the attraction to white, blue, green and red LED lights revealed attraction to each of these light types in newborns, but no significant attraction in adult females. Both newborns and adult females exhibited reduced mobility under red LED light. In a preference experiment in which all light colours were presented simultaneously, newborns were most attracted to the white light (52.0% of the newborns) and least attracted to the red light and darkness (6.8% and 6.7% of the newborns respectively). These results support the potential efficacy of using white light to attract newborns into the baskets in guppy breeding tanks and reduce cannibalism effects.     

Antioxidant defence to ammonia stress of characins (Hyphessobrycon eques Steindachner) fed diets supplemented with carotenoids

This study aimed to find out whether dietary carotenoid (CD) supplement could influence the resistance of characins (Hyphessobrycon eques Steindachner) to ammonia stress. Two types of CD and its combination [astaxanthin (AX), β‐carotene (BC), 1 : 1 combination of AX and BC (MX)] at three concentrations (10, 20 and 40 mg kg⁻¹) were used resulting in nine pigmented diets. No differences in growth and survival of the fish among treatments were found after 8‐week rearing. Experimental and control fish were then exposed to 15 mg total ammonia nitrogen L⁻¹ (stress group) and 0.15 mg total ammonia nitrogen L⁻¹ (normal group) for 72 h, and their blood was withdrawn. No mortality resulted under such TAN concentrations. Serum total antioxidant status (TAS), serum antioxidant enzymes [superoxide dismutase (SOD), glutathione peroxidases (GPx)] and serum transaminases [alanine aminotransferase (ALT), aspartate aminotransferase (AST)] were chosen as indices of fish antioxidant capacity or stress resistance. SOD, GPx and AST were affected by the interactions of dietary CD and ammonia stress. The activities of TAS, SOD, GPx and AST increased under the stress. Dietary CD reduced serum SOD, GPx, ALT and AST activities. In conclusion, dietary CD increased the resistance of characins to ammonia stress.     

Dietary supplementation of β‐glucan improves growth performance,the innate immune response and stress resistance of red sea bream,Pagrus major

A 56‐day feeding trial was conducted to evaluate the effects of supplemented diets with β‐glucan (BG) at four levels [0 (D1), 250 (D2), 500 (D3) and 1000 (D4) mg BG kg^?1] on red sea bream, Pagrus major. The obtained results revealed a significant increase (P < 0.05) in final body weight, weight gain, specific growth rate, feed intake, body protein content, lysozyme activity and tolerance against low‐salinity stress test in all BG‐supplemented groups when compared with BG‐free group. Furthermore, D4 group resulted in a significant increase in feed efficiency ratio, protein gain, protein and lipid digestibilities, serum bactericidal activity and peroxidase content when compared with D1 group (P < 0.05). Haematocrit and plasma protein content in D3 group were significantly higher than those in D1 group (P < 0.05). Interestingly, BG supplementation decreased glutamic oxaloacetic transaminase (GOT) in D2 group and reactive oxygen metabolites in D2, D3 and D4 groups when compared with D1 group. Following low‐salinity stress test, significantly higher amounts of secreted mucus were observed in fish fed D2 and D4 diets than those from fish fed D1 diet (P < 0.05). In conclusion, the supplementation of BG improves growth, stress resistance and immune response of P. major.     

The antioxidant capacity response to hypoxia stress during transportation of characins (Hyphessobrycon callistus Boulenger) fed diets supplemented with carotenoids

This study aimed to determine whether dietary carotenoid (CD) supplements could affect the antioxidant capacity of characins Hyphessobrycon callistus upon hypoxia stress at live transportation. Two types of CD [astaxanthin (AX), β‐carotene (BC)] and their 1:1 combination (MX) at three concentrations (10, 20 and 40 mg kg^?1) were supplemented, resulting in nine CD diets. After 8 weeks' rearing, the resulting fish were divided into two subgroups and exposed to hypoxia or normoxia. Hypoxia involved a gradual decrease in dissolved oxygen (DO) from 6.5 to <1.0 mg L^?1. Normoxia was DO kept in saturation. Hypoxia led to an increase in the total antioxidant status (TAS), superoxide dismutase (SOD), glutathione peroxidases (GPx) and aspartate aminotransferase (AST) activity of blood serum in fish, but had no effect on alanine aminotransferase (ALT). Under hypoxia, fish fed CD diets had lower SOD, GPx and ALT activity than control fish, showing that dietary CD could increase the antioxidant capacity and protection of the liver. Dietary AX was more effective for antioxidant capacity than BC and MX when under hypoxia stress, because GPx, ALT and AST were lower in AX‐fed fish. Except TAS, the other four enzyme activities showed decreasing trends with increasing dietary CD concentrations.     

Effect of supplementation of dietary fibre concentrates on biochemical parameters,stress response,immune response and skin mucus of jundiá (Rhamdia quelen)

The aim of this study was to assess the effects of different dietary fibre concentrates (DFC: Mucilage = MG; pectin = PN or β‐glucan+mannan = βg+M), on biochemical parameters, stress and immune response and skin mucus of jundiá (Rhamdia quelen). The fish (7.16 ± 0.06 g) were fed with Control diet (0 g/kg of DFC); diet supplemented with 5 g/kg of commercial prebiotic (CP 5) or diets supplemented with 5 or 10 g/kg of MG; PN or βg+M. After 8 weeks of the feeding trials, biochemical parameters (cholesterol, glucose, albumin and total protein), cortisol, immunoglobulin IgM and mucoproteins of skin mucus were assessed. Results demonstrated that supplementation with PN increased cholesterol levels (p<.05). After application of the stressor, most fish, except those in the PN and 10 g/kg MG groups, showed significant increases (p<.05) in cholesterol, glucose and albumin levels. The jundiás showed no difference in cortisol levels after application of the stressor (p>.05). IgM levels were significantly high in fish supplemented with DFC (p<.05). However, the concentration of mucoproteins in skin mucus was not influenced in the different treatments (p>.05). The results showed that supplementation with DFC promoted beneficial effects on the metabolism of jundiá.     

Dietary α‐linolenic acid affects lipid metabolism and tissue fatty acid profile and induces apoptosis in intraperitoneal adipose tissue of juvenile grass carp (Ctenopharyngodon idella)

A 56‐day feeding trial was conducted to elucidate the effects and mechanism action of dietary α‐linolenic acid (ALA, 18:3n‐3) on lipid accumulation and fatty acid profile of muscle, hepatopancreas and intraperitoneal fat (IPF) in juvenile grass carp using three isonitrogenous and isoenergetic semi‐purified diets containing 0.0% (control group), 1.0% and 2.0% ALA, respectively. The lowest intraperitoneal fat (IPF) ratio was found in 2.0% group. In the muscle, hepatopancreas and IPF, docosahexaenoic acid (DHA, 22:6n‐3) and eicosapentaenoic acid (EPA, 20:5n‐3) contents increased with the increase in dietary ALA. In the IPF, caspase 3, caspase 8 and caspase 9 showed the highest activities in 2.0% group, while the value of Bcl‐2/Bax (B‐cell leukaemia 2/Bcl‐2‐associated X protein) reached the lowest. Meanwhile, swelling of the IPF mitochondria was observed in 2.0% group. The gene expressions of fatty acid desaturase (FAD) and fatty acid elongase (ELO) in the hepatopancreas and muscle showed significantly higher levels in the treatment groups, whereas an opposite trend was existed in the IPF. Fatty acid synthase (FAS), sterol regulatory element binding protein‐1c (SREBP‐1c) in the IPF and hepatopancreas reached the lowest in 2.0% group. Overall, dietary ALA could promote n‐3 highly unsaturated fatty acids (HUFAs) synthesis and suppress the accumulation of lipid by decreasing the expression of related genes and promoting the apoptosis in IPF.     

Dietary supplementation with biofloc promotes growth,improves immune and antioxidant status,and upregulates NF‐κB/Nrf2 signalling molecules and stress resistance in Rhynchocypris lagowskii Dybowski

Bioflocs are rich in a variety of probiotics and bioactive compounds, which have been documented to promote growth, regulate antioxidant status and enhance the immune system. A 60‐day experiment was conducted to investigate the effects on growth, antioxidant and immune status, NF‐κB/Nrf2 signalling molecules and stress resistance in juvenile Rhynchocypris lagowskii Dybowski fed different dietary biofloc supplementations. Four hundred fifty R. lagowskii were fed five experimental diets containing graded levels of biofloc from 0 to 160 g/kg (referred to as B0, B4, B8, B12 and B16, respectively). The results showed that biofloc supplementation markedly promoted growth performance (weight gain rate, specific growth rate); improved amylase, lipase and protease activity; and enhanced the activities of glutathione peroxidase, catalase, total superoxide dismutase, total antioxidant capacity, lysozyme, complement C3, complement C4 and immunoglobulin M in the gill, hepatopancreas, kidney, gut and serum. The malondialdehyde content was significantly reduced with treatment B12. In addition, the levels of heat shock protein 70, heat shock protein 90, interferon‐γ and insulin‐like growth factor I were substantially increased with treatment B12 compared to those with the control treatment. Furthermore, biofloc assisted in regulating the expression of NF‐κB/Nrf2 signalling molecule genes, including NF‐κB, TNF‐α, IL‐1β, IL‐8, IL‐10, TGF‐β, Nrf2, Keap1, Maf, HO‐1, CAT, GPX, GCLC and CuZn‐SOD. Overall, our results suggest that dietary supplementation with biofloc can promote growth, improve immune and antioxidant status, and enhance NF‐κB/Nrf2 signalling molecule expression and stress resistance in juvenile R. lagowskii. A suitable supplementation level of approximately 120 g/kg biofloc is recommended in the present study.     

Dietary microbial levan ameliorates stress and augments immunity in Cyprinus carpio fry (Linnaeus, 1758) exposed to sublethal toxicity of fipronil

A 45‐day feeding trial was conducted to study the stress ameliorating and immunomodulatory role of microbial levan in Cyprinus carpio fry exposed to sublethal dose (1/10th LC₅₀) of fipronil [(±)‐5‐amino‐1‐(2,6‐dichloro‐α,α,α‐trifluoro‐p‐tolyl)‐4‐trifluoromethylsulfinylpyrazole‐3‐carbonitrile]. Two hundred and twenty‐five fry were randomly distributed in five treatments in triplicates. Four purified diets were prepared with graded levels of microbial levan. Five different treatment groups were levan control L₀P₀ (basalfeed + 0% levan without exposure to pesticide); pesticide control L₀P₁ (basalfeed + 0% levan with exposure to pesticide); L_0.25P₁ (basalfeed + 0.25% levan with exposure to pesticide); L_0.50P₁ (basalfeed + 0.50% levan with exposure to pesticide) and L_0.75P₁ (basalfeed + 0.75% levan with exposure to pesticide). Lactate dehydrogenase (LDH), malate dehydrogenase (MDH) and fructose‐1,6‐diphosphatase (FDPase) activites were significantly (P < 0.05) increased, whereas alkaline phosphatase (ALP), adenosine triphosphatase (ATPase) and acetyl choline esterase (AchE) activities were significantly (P < 0.05) reduced in higher levan‐fed groups. RBC, haemoglobin and WBC counts were significantly (P < 0.05) increased in the levan‐fed groups. Similar trends were also observed for the total serum protein, globulin, NBT and lysozyme activities. Blood glucose and serum cortisol exhibited a third order polynomial relationship with increasing level of dietary levan. Overall result showed stress ameliorating, immunostimulating and protective role of microbial levan against fipronil‐induced stress in C. carpio fry at 0.75% level of dietary levan supplementation.     

Dietary effects of poly‐β‐hydroxybutyrate on the growth performance,digestive enzyme activity,body composition,mineral uptake and bacterial challenge of rainbow trout fry (Oncorhynchus mykiss)

This study investigated the effects of diet supplementation with poly‐β‐hydroxybutyrate (PHB) on growth performance, digestive enzyme activity, lipid metabolism, mineral uptake and bacterial challenge of the rainbow trout fry (initial weight: 111.3 ± 13.7 mg) during a 6‐week experimental period. In the experimental set‐up, the commercial diet of rainbow trout fry was replaced with 0.5%, 1% and 2% PHB. The results of our work showed that the replacing of diets with PHB in experimental treatments decreases the weight of rainbow trout fry during the first 2 weeks before significantly increasing final weight at the end of the 6‐week period. PHB also improved digestive enzyme activity in experimental treatments. The highest total protease, pepsin activity and pancreatic enzyme secretion were observed with the 0.5% PHB treatment. A higher concentration of Na and K was observed in the whole body of the fry fed on 1% and 2% PHB‐supplemented diets. Enhanced survival rates occurred in all groups of fry after bath exposure to Yersinia ruckeri compared to rates in those fed the control diet. Our results suggest that the diet supplemented with PHB may improve growth performance, digestive enzyme activity and the functioning of the immune system. These positive effects could be considered for new applications in aquaculture.     

Improved vitellogenesis,gonad development and egg diameter in catfish (Pangasianodon hypophthalmus) supplemented with turmeric (Curcuma longa) powder

An experiment was conducted to study the effect of supplementing catfish (Pangasianodon hypophthalmus) with turmeric (Curcuma longa) on the vitellogenic capacity of the liver as a new method to improve reproductive performance in oviparous animals. The experimental catfish were assigned to a completely randomized design consisting of four doses of supplemental turmeric (0, 1.2, 2.4 and 4.8 g/kg feed) with 10 catfish with body weights ranging from 2 to 4 kg in each group. Blood samples were collected on days 0, 14, 28, 42 and 56 of turmeric supplementation to measure the concentrations of plasma oestradiol‐17ß and vitellogenin. The weights of the body, gonads and liver were measured on days 0, 28 and 56. The fecundities of the catfish were measured on days 28 and 56 of turmeric supplementation, and egg diameters were measured on days 28 and 42. The results showed that catfish supplemented with turmeric at a dose of 2.4 or 4.8 g/kg feed had higher growth rates, higher plasma oestradiol‐17β and vitellogenin concentrations, and higher gonad somatic index values and egg diameters. This is the first report demonstrating that the hepatoprotective activity of the curcumin in turmeric could be used to improve both vitellogenin synthesis, which improves nutrient deposition in the ovulating eggs, and the reproductive performance of teleost fish and oviparous animals.     

Effect of different dietary microalgae combinations on growth and survival of black‐lip pearl oyster (Pinctada margaritifera) larvae and the feasibility of replacing microalgae with a dietary lipid emulsion

A 10‐day experiment was performed to examine different mono, binary and ternary dietary combinations on survival and growth of D‐shaped and umbone black‐lip pearl oyster, Pinctada margaritifera, larvae. The three tropical microalgae species were the flagellate Isochrysis galbana clone T. Iso (CS‐177) and diatoms Chaetoceros calcitrans (CS‐178) and Chaetoceros muelleri (CS‐176) which were fed to D‐shaped and umbone larvae at a density of 7000 and 14 000 cells mL^?1, respectively. A second experiment was performed to investigate the feasibility of replacing T. Iso with a lipid emulsion for both D‐shaped and umbone larvae for 10 and 12 days, respectively. The treatments included only T. Iso, unfed and lipid emulsion to substitute T. Iso at levels of 10% (LIP10), 30% (LIP30) and 100% (LIP100). In the first experiment, results showed that a monospecific diet of T. Iso led to significantly higher (P < 0.05) survival and growth of D‐shaped larvae than all the other treatments. Meanwhile, D‐shaped larval survival was significantly lower when only fed C. calcitrans as well as growth for those fed C. calcitrans or in combination with C. muelleri. However, for umbone larvae, survival and growth were significantly higher when fed a binary combination of T. Iso and C. muelleri or the ternary combination of T. Iso, C. muelleri and C. calcitrans compared with all other treatments. For the second experiment, results showed that with increasing lipid emulsion replacement, survival of both D‐shaped and umbone larvae significantly decreased (P < 0.05); however, the LIP100 treatment was not significantly different (P > 0.05) from the unfed treatment. For D‐shaped larvae, no significant growth difference was detected (P > 0.05) between the T. Iso and LIP10 fed treatments while for umbone larvae, the T. Iso, LIP10 and LIP30 were not significantly different (P > 005). These results indicate that microalgae combinations appear more necessary for later staged P. margaritifera larvae. In addition, the use of a lipid emulsion appeared to provide some nutrition to the larvae, although more research should be conducted to improve the use of such replacements.     

Comparative analysis of the predominant carotenoids and chemical components in the common and orange‐muscle mutant of Haliotis gigantea

The Xishi abalone (Haliotis gigantea) is an economically significant aquaculture species in southern China. We identified a novel orange‐muscle mutation present in less than 2% of the cultured population. High‐performance liquid chromatography and electrospray ionization mass spectrometry identified zeaxanthin and β‐carotene as the two main pigments present in the orange‐muscle abalone. The concentrations of zeaxanthin and β‐carotene detected in the orange‐muscle abalones were significantly higher than those detected in the common abalones (p < 0.01). Notably, the concentration of zeaxanthin was approximately 16.5 times higher in orange‐muscle abalones than in common abalones, which was also the species with highest carotenoid content reported in shellfish to date. Furthermore, the contents of total amino acids and essential amino acids in the orange‐muscle abalone were higher than that in the common abalone, though not significantly, while the content of non‐essential amino acids was significantly higher in the orange‐muscle abalone than that in the common abalone (p < 0.05). The saturated fatty acid content in the orange‐muscle abalones was slightly lower than that in the common abalones, and the unsaturated fatty acid content was higher in the orange‐muscle abalones than that in the common abalones. These results could provide a basis for characterizing the mechanism by which carotenoids accumulate in abalones and further guide the breeding of abalone with orange‐muscle mutants in the future.     

Intra‐annual changes in reproductive indices of male and female Himalayan snow trout,Schizothorax richardsonii (Gray, 1832)

Periodic changes in reproductive hormone levels, gonadal histology and gonadosomatic index (GSI) of snow trout, Schizothorax richardsonii, were examined to ascertain annual cycle of gonadal development and reproductive status in their natural habitat. In females, there were coherent changes in plasma 17β‐oestradiol and vitellogenin along with GSI, oocyte maturation and vitellogenic progression, collectively indicating two distinct maturation peaks during the months of September and February. Coinciding with this, in males, plasma 11‐keto testosterone was also noticeably higher during September and February, with highest GSI values in September. However, plasma 17α, 20β‐dihydroxyprogesterone levels in males were found to be persistently high from September to February. This observation suggests the potential presence of matured oozing males over a longer period, unlike in females. Overall, the close association between reproductive hormone levels, GSI and gonadal maturation stages in males and females (particularly, the presence of postovulatory follicle complexes) with apparent natural synchronization clearly indicates that S. richardsonii breeds twice in a year, possibly during late September to early November and late February to early April in the coldwater riverine habitats of the Indian Himalayan region.     

Effects of dietary β‐carotene levels on growth and liver vitamin A concentrations of the soft‐shelled turtle,Pelodiscus sinensis (Wiegmann)

A feeding trial was conducted to evaluate the effect of dietary β‐carotene level on the growth and liver vitamin A concentrations in soft‐shelled turtles, Pelodiscus sinensis, fed a vitamin A‐free diet. Soft‐shelled turtles were fed diets containing 0, 14.5, 26.5, 47.5, 87.3, 112.8 and 163.8 mg β‐carotene kg^?1 for 10 weeks. Although it was not statistically significant due to high deviation within each group, mean weight gain of soft‐shelled turtles fed the diet without β‐carotene supplementation was the lowest among all test groups. Vitamin A concentrations in liver of turtles significantly (P<0.05) increased when dietary β‐carotene level reached 47.5 mg kg^?1 indicating that soft‐shelled turtles were capable of converting β‐carotene to vitamin A. Analysed by regression modelling, dietary β‐carotene levels for optimal growth and maximal liver vitamin A contents of juvenile soft‐shelled turtles fed the vitamin A‐free diets were 49.1 and 88.7 mg kg^?1 respectively.     

Combination effects of dietary EPA and DHA plus alpha‐tocopherol: effects on performance and physiological status of Caspian brown trout (Salmo trutta caspius) fry

The effects of dietary n‐3 highly unsaturated fatty acids [eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA)] with α‐tocopherol on growth, non‐specific immune response and oxidative status were investigated in Caspian brown trout, Salmo trutta caspius, fry. Six experimental diets containing three different dietary levels of n‐3 HUFAs (low: 1 + 0.5% of total fatty acids, DHA+ EPA, medium: 2 + 1%, DHA + EPA, high: 4 + 2%, DHA + EPA) with two different levels of α‐tocopherol (low: 300 and high: 1000 mg kg^?1 diet) were prepared and named: LL, LH, ML, MH, HL and HH (HUFA/α‐tocopherol) groups, respectively. Diets were fed to triplicate groups of 60 fry with an initial weight of 600 ± 25 mg for 10 weeks. Results showed that increase in dietary DHA and EPA up to high level improved fry growth in terms of the body weight and specific growth rate, particularly when dietary α‐tocopherol levels were high, suggesting a higher antioxidant protection value when these fatty acids are high. At all dietary n‐3 HUFA levels, increase in α‐tocopherol from low to high level enhanced the alternative complement (ACH50) activity. Fry fed diets medium and high n‐3 HUFA displaying significantly higher lysozyme activity (P < 0.05). Moreover, fish fed medium or high levels of n‐3 HUFA had significantly lower prostaglandin E2 (PGE2) than those fed low n‐3 HUFA (P < 0.05). Significant differences in antioxidant enzymes (catalase, glutathione peroxidase, glutathione S‐transferase, glutathione reductase and superoxide dismutase) activity were also observed between groups, with higher activity in high levels of n‐3 HUFA (P < 0.05). Results of this study suggest that the effect of dietary n‐3 HUFA on examined non‐specific immunity parameters are not uniform; however, these impacts are closely related to the α‐tocopherol supplement and their interaction. In conclusion, increased dietary levels of n‐3 HUFA and α‐tocopherol would enhance growth performance and welfare of this species.