Sperm density, seminal plasma composition and their physiological relationship in the endangered Caspian brown trout (Salmo trutta caspius)

Sperm density, mineral and organic composition of the seminal plasma and their physiological relationship were investigated in the Caspian brown trout (Salmo trutta caspius). To establish a rapid and accurate method for assessment of sperm density, three different techniques were used: sperm counting, spectrophotometry (at 480 nm) and determination of the spermatocrit. The seminal plasma contained 159.26±8.84 mM sodium (Na), 33.72±2.01 mM potassium (K), 133.04±5.96 mM chlorine (Cl), 1.68±0.2 mM calcium (Ca) and 0.988±0.13 mM magnesium (Mg). The following organic components were found: total protein 0.75±0.14 mg 100 mL⁻¹, cholesterol 2.86±0.58 mg L⁻¹ and glucose 3.81±1.04 mM L⁻¹. The mean sperm density was estimated to be 3.3 × 10⁹ spermatozoa mL⁻¹. The spermatocrit (%) ranged from 25 to 52 in sperm samples. Highly significant linear relationships were found between sperm density and spermatocrit (R²=0.703, P<0.001) and sperm density and optical density (R²=0.909, P<0.001), indicating that optical density can be used as a quick and accurate method of estimating sperm density. Significant relationships were also found between sperm density and Ca, Mg and total protein of seminal plasma. A significant correlation was also observed between the Ca and Mg concentrations (R²=0.774, P<0.01). The following correlations were observed between mineral and organic components: total protein and Ca (R²=0.462, P<0.05), total protein and Mg (R²=0.518, P<0.05) and glucose and Cl (R²=0.374, P<0.05). These parameters should be considered when developing procedures for either artificial fertilization or for cryopreservation of sperm.     

Antibiotics,Penicillin and Sreptomycin improve semen quality indices of endangered Caspian brown trout,Salmo trutta caspius (Kessler, 1870) during in vitro short‐term storage

In this study, we examined the effects of 500 IU mL^?1 penicillin + 500 μg mL^?1 streptomycin sulphate on semen quality indices of endangered caspian brown during 12 days short‐term storage at 4°C. Twenty‐four millilitre semen samples with good quality were considered for the experiment. The semen samples were then stored in the presence and absence of 500 IU mL^?1 penicillin + 500 μg mL^?1 streptomycin sulphate. The semen quality parameters including percentage and duration of sperm motility were measured 0, 3, 6, 9 and 12 days after storage. In the antibiotic receiving group, the values of percentage and duration of sperm motility reduced 3 and 6 days after storage respectively and reduced to lowest levels at day 12. In the antibiotic‐free group, the duration and percentage of sperm motility decreased significantly after 3 days of storage and reached to lowest values at day 12. Also, percentage and duration of sperm motility in each storage time were significantly higher in the antibiotic receiving group than in the antibiotic‐free group. The overall values of percentage and duration of sperm motility for all storage periods were higher in the antibiotic receiving group than in the antibiotic‐free group. In conclusion, our results demonstrated that 500 IU mL^?1 penicillin + 500 μg mL^?1 streptomycin sulphate improves the viability of caspian brown trout during short‐term storage.     

Effects of spirulina powder (Spirulina platensis) as a dietary additive on Oscar fish,Astronotus ocellatus: Assessing growth performance,body composition,digestive enzyme activity,immune‐biochemical parameters,blood indices and total pigmentation

In this study, the effect of spirulina meal (Spirulina platensis) as a feed additive on growth and physiological response of Oscar fish,Astronotus ocellatus, was assessed using four diets including control without spirulina supplementation, 13.75 g/kg spirulina (SP1), 27.50 g/kg spirulina (SP2) and 55 g/kg spirulina (SP3). One hundred and twenty fish (10.41 ± 0.25 g) were disturbed to twelve aquariums and fed to apparent satiation three times a day for 8 weeks. The result showed that the SP3 treatment had statistically better growth performance, feeding parameters, total protease activity, total protein and lipid contents than the control. Moreover, haemoglobin, haematocrit, mean erythrocyte haemoglobin and mean erythrocyte haemoglobin concentrations were improved considerably in the SP3 treatment compared with the control. The SP3 treatment showed higher total protein and albumin levels, but lower triglycerides, cholesterol levels and liver enzyme activities than the control. Also, serum complements (C3, C4), lysozyme activity and total carotenoid contents in fish skin significantly increased in the SP3 treatment compared with the control. The results of this study suggest that spirulina powder at the level of 55 g/kg of the diet can be used as a functional natural feed additive to improve the performance of Oscar fish.     

Effect of partial replacement of dietary fish meal by soybean meal with betaine attractant supplementation on growth performance and fatty acid profiles of juvenile rainbow trout (Oncorhynchus mykiss)

This study was conducted to investigate the effects of dietary incorporation of soybean meal for fish meal replacement and supplementation of betaine as an attractant on growth performance and fatty acid profiles of rainbow trout (Oncorhynchus mykiss). Juvenile trout were fed practical diets, with increasing soybean levels and betaine supplementations. The experimental diets consisted of a control diet with fish meal as the sole protein source (control) and no attractant, 25% soybean‐1% betaine diet (SBM25‐B1), 50% soybean‐2% betaine diet (SBM50‐B2), and 50% soybean‐0% betaine diet (SBM50‐B0). Average body weight was 12.69 ± 0.16 g at the beginning of experiments. Following 54 days feeding programme with experimental diets, average body weights reached 47.45 ± 1.22 g, 58.11 ± 1.77 g, 56.34 ± 1.87 g and 53.76 ± 1.74 g in the control, SBM25‐B1, SBM50‐B2 and SBM50‐B0 groups respectively. As compared with control treatment, significant differences were observed in weight gain, specific growth rate and feed intake of 1% betaine treatment at 25% soybean‐meal‐incorporated diet (p < .05), but no differences were observed in feed conversation ratio and survival rates (p > .05). Compared with the control treatment, betaine‐supplemented groups had significantly higher total saturated fatty acid contents (p < .05). On the other hand, the control treatment showed a significantly higher level of monounsaturated fatty acid than the betaine‐supplemented groups (p < .05). Significant differences were observed in fatty acid profile of 1% betaine‐supplemented group (p < .05) compared with the control. Present findings revealed that 1% betaine supplementation with dietary incorporation of soybean meal at 25% level positively influenced growth performance, feed utilization and fatty acid profiles of rainbow trout juveniles.     

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.     

The effects of Spirulina platensis meal on proximate composition,fatty acid profile and lipid peroxidation of rainbow trout (Oncorhynchus mykiss) muscle

This study evaluated the effects of diets containing 0, 25, 50, 75 and 100 g kg^?1 Spirulina platensis on proximate composition, fatty acid profile and lipid peroxidation of rainbow trout (Oncorhynchus mykiss). Supplementation of S. platensis did not change moisture and protein contents, but fish fed 50 and 100 g kg^?1 S. platensis had lower muscle lipid content than those fed control diet (P < 0.05). Fish fed 100 g kg^?1 of S. platensis contained lower percentages of saturated and monounsaturated fatty acid and a higher percentage of polyunsaturated fatty acid than those fed control diet (P < 0.05). The n‐3/n‐6 ratio of the fatty acid increased and muscle atherogenic and thrombogenic indices were significantly decreased as the dietary supplement of S. platensis increased. Furthermore, lipid peroxidation of the fillet significantly decreased with increasing dietary S. platensis at 4 °C and at ?20 °C (P < 0.05). The results of this study show that supplementation of S. platensis to the diet improves muscle quality of the rainbow trout.     

Effects of total fish oil replacement to vegetable oils at two dietary lipid levels on the growth,body composition,haemato‐immunological and serum biochemical parameters in caspian brown trout (Salmo trutta caspius Kessler, 1877)

This research aimed to evaluate the effects of two dietary fat levels [low fat (LF) (10%), high fat (HF) (20%)] and sources [fish oil (FO), vegetable oil (VO)] on the growth and some physiological parameters of Caspian brown trout fingerlings for 60 days. Tuna oil or blends of canola and soybean oils (85:15) were added to diets to design four feeds namely LFFO, HFFO, LFVO and HFVO according to the fat levels and sources. The fish fed the LFFO diet had lower weight gain than the other fish (P<0.05). The total n‐6 fatty acids increased in fish fed diets with the blends of VO, while the total n‐3 fatty acids decreased in these fish (P<0.05). Serum lysozyme activity was higher in fish fed the HFVO diet than the other fish (P<0.05). Serum glucose, total cholesterol, triglyceride and very low‐density lipoprotein were lower in fish fed LFFO than the other fish (P<0.05). The present study demonstrates that in terms of fish growth, VOs can be used as an alternate source of dietary fat, whereas fish health and nutritional value are improved with the LFFO diet. According to these results, a partial substitution of FO by VO in high‐level fat diets is suggested for long‐term feeding of Caspian brown trout.     

Partial and total replacement of fish meal by marine microalga Spirulina platensis in the diet of Pacific white shrimp Litopenaeus vannamei: Growth,digestive enzyme activities,fatty acid composition and responses to ammonia and hypoxia stress

In the this study, we evaluated the effect of replacement of fish meal by a marine microalgae Spirulina platensis on growth, digestive enzyme activities, fatty acid composition and responses to ammonia and hypoxia stress in Pacific white shrimp Litopenaeus vannamei (2.6 ± 0.2 g). Experimental diets contained S. platensis at 0%, 25%, 50%, 75% and 100% replacement levels. After 8 weeks of feeding trial, growth parameters and proximate body composition were not significantly different among treatments (p > .05). Amylase and lipase activities did not show any significant differences between control group and other experimental diets (p > .05), while activities of trypsin and chymotrypsin were significantly higher in shrimp fed diet with 50% substitution of microalgae compared to control group. Fatty acid contents, particularly polyunsaturated fatty acids (PUFAs) including arachidonic acid (ARA), docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA), were significantly higher in control diet compared to other experimental diets. On the contrary, the majority of fatty acids including the contents of PUFAs in the whole body of L. vannamei fed with different levels of S. platensis were significantly higher compared to those of control group. After 48‐h exposure to ammonia, survival per cent was not statistically different between all groups (p > .05), but in hypoxia challenge, the survival per cent of control group was significantly less than that of treatments fed diets contained S. platensis (p < .05). Altogether, o ur results demonstrated the effectiveness of S. platensis as a reliable protein source for substitution of fish meal in shrimp aquaculture.     

Effects of dietary n‐3HUFA on different growth stages of the white shrimp,Litopenaeus vannamei: Growth,haematological characteristics,enzyme activities and fatty acid profiles

This study investigated the effects of n‐3 high unsaturated fatty acid (n‐3HUFA) levels on the growth performance, antioxidant enzyme activities and fatty acid profiles of both subadult and adult Litopenaeus vannamei (L. vannamei). Seven iso‐nitrogenous and iso‐lipidic diets were used, containing n‐3HUFA concentrations of 1.6 (control), 4.8, 7.4, 13.9, 23.9, 29.2 and 34.4 g/kg, respectively. Two 8‐week feeding trials were conducted to determine the dietary n‐3HUFA requirements of L. vannamei with an initial body weight of 4.25 ± 0.00 g (subadults) and 8.50 ± 0.01 g (adults). The results showed that the dietary n‐3HUFA level significantly affected the weight gain rate (WGR), specific growth rate, the feed conversion ratio and the hepatosomatic index (HSI) (p < 0.05), but did not significantly affect the survival rate (p > 0.05). At appropriate level, dietary n‐3HUFA improved growth performance and HSI of both subadult and adult L. vannamei. Both subadults and adults showed significant differences in body composition (p < 0.05), except for moisture and crude ash (p > 0.05). Cholesterol and low‐density lipoprotein significantly decreased with increasing dietary n‐3HUFA both in subadults and adults (p < 0.05); however, triglyceride showed no significant change (p > 0.05). High‐density lipoprotein (HDL) in subadults was significantly affected by dietary n‐3HUFA (p < 0.05), but followed no apparent regularity; HDL significantly changed in adults and showed an upward trend followed by a downward trend (p < 0.05). There was no significant effect on aspartate transaminase (AST) activity in subadults, but AST in adults and alanine transaminase (ALT) in subadults and adults were significantly affected (p < 0.05). Dietary n‐3HUFA significantly affected serum polyphend oxidase, malic dehydrogenase, alkaline phosphatase, superoxide dismutase and sodium‐potassium adenosine triphosphatase enzyme activities in gills (p < 0.05). The fatty acid composition of the shrimp tissue was associated with the fatty acid composition of the diet. Dietary n‐3HUFA supplementation significantly improved the contents of tissue ∑HUFA and n‐3HUFA, increased the n‐3/n‐6 ratio in the tail muscle and decreased the contents of tissue polyunsaturated fatty acid and saturated fatty acid (p < 0.05). Based on the WGR, the broken‐line equations indicated that the optimum requirements of dietary n‐3HUFA were determined to be 9.0 and 5.1 g/kg for subadult and adult L. vannamei, respectively.