Integration Process for Protein Extraction from Microalgae Using Liquid Biphasic Electric Flotation (LBEF) System

Microalgae are the most promising sources of protein, which have high potential due to their high-value protein content. Conventional methods of protein harnessing required multiple steps, and they are generally complex, time consuming, and expensive. Currently, the study of integration methods for microalgae cell disruption and protein recovery process as a single-step process is attracting considerable interest. This study aims to investigate the novel approach of integration method of electrolysis and liquid biphasic flotation for protein extraction from wet biomass of Chlorella sorokiniana CY-1 and obtaining the optimal operating conditions for the protein extraction. The optimized conditions were found at 60% (v/v) of 1-propanol as top phase, 250 g/L of dipotassium hydrogen phosphate as bottom phase, crude microalgae loading of 0.1 g, air flowrate of 150 cc/min, flotation time of 10 min, voltage of 20 V and electrode’s tip touching the top phase of LBEF. The protein recovery and separation efficiency after optimization were 23.4106?±?1.2514% and 173.0870?±?4.4752%, respectively. Comparison for LBEF with and without the aid of electric supply was also conducted, and it was found that with the aid of electrolysis, the protein recovery and separation efficiency increased compared to the LBEF without electrolysis. This novel approach minimizes the steps for overall protein recovery from microalgae, time consumption, and cost of operation, which is beneficial in bioprocessing industry.     

Optimizing a continuous flow lipid extraction system (CFLES) used for extracting microalgal lipids

A laboratory‐made continuous flow lipid extraction system (CFLES) was devised to extract lipids from microalgae Nannochloropsis sp., a potential feedstock for biodiesel fuel, with a focus to assess the workable temperatures and pressures for future industrial applications. Using conventional solvents, the CFLES recovered 100% of the lipids extracted with conventional Soxhlet extraction. The optimum temperature and pressure were found to be 100 °C and 50 psi, respectively; conditions significantly lower than those normally used in pressurized liquid extractions requiring specialized equipment. Approximately 87% of the extracted oil was successfully transesterified into biodiesel fuel (fatty acid methyl esters). Preliminary calculations based on the tested lab‐scale system indicated savings in energy, solvent consumption, and extraction time as 96%, 80%, and more than 90%, respectively, as compared to Soxhlet extraction. However, the true cost savings can only be assessed at scaled up level. Energy efficiency of CFLES was calculated as 48.9%. Residual water (~70%) in the biomass had no effect on the extraction performance of CFLES, which is expected to help the process economics at scaled up application. The effect of temperature and pressure on the fatty acids profile of Nannochloropsis sp. is also discussed. Based on the existing literature, the authors believe that a pressurized liquid extraction system with continuous solvent flow has not been reported for lipid extraction from Nannochloropsis sp.     

Pressurized extraction of unsaturated fatty acids and carotenoids from wet Chlorella vulgaris and Phaeodactylum tricornutum biomass using subcritical liquids

The objective of this study was to investigate the extraction of lipids, for example, mono‐ and polyunsaturated fatty acids (PUFA) as well as carotenoids, from wet microalgae biomass using pressurized subcritical extraction solvents, which meet the requirements of food and feed applications. To demonstrate the effect of the solvent and temperature on the lipid yield, we chose two microalgae species, viz. Chlorella vulgaris and Phaeodactylum tricornutum, differing in their biochemical composition fundamentally. In case of P. tricornutum, ethanol showed the highest fatty acid yield of 85.9% w/w. In addition to eicosapentaenoic acid (EPA), the ethanolic extracts contained exceptional amounts of fucoxanthin (up to 26.1 mg/g d. w.), which can be beneficial to protect unsaturated fatty acids from oxidation processes and in terms of human nutrition. For C. vulgaris, a fatty acid yield of 76.5% w/w was achieved from wet biomass using ethyl acetate at 150°C. In general, an increase in the extraction temperature up to 150°C was found to be important in terms of fatty acid yield when extracting wet microalgae biomass. The results suggest that it is possible to efficiently extract both fatty acids and carotenoids from wet microalgae by selecting suitable solvents and thus circumvent energy‐intensive drying of the biomass.     

Dietary niacin requirement of fingerling Channa punctatus (Bloch)

A 16‐week experiment was accomplished to determine the dietary niacin requirement of fingerling Channa punctatus (6.8 ± 0.92 cm; 4.65 ± 0.46 g) by feeding seven casein‐gelatin based isonitrogenous (450 g/kg CP) and isoenergetic (18.39 kJ/g GE) diets with graded levels of niacin (0, 10, 20, 30, 40, 50 and 60 mg/kg diet) twice a day to apparent satiation to triplicate groups of fish. Significantly best absolute weight gain (AWG; 38.19 g/fish), feed conversion ratio (FCR; 1.42) and protein retention efficiency (PRE; 26.47%) were registered in fish fed 40 mg niacin/kg diet. Also, fish fed above diet exhibited maximum carcass protein. Hemoglobin (Hb), RBCs counts and hematocrit (Hct) were improved with the incremental levels of dietary niacin up to 40 mg/kg. However, liver niacin content showed the positive correlation up to 50 mg/kg niacin and then leveled off. Fingerling C. punctatus fed niacin‐free diet showed retarded growth, poor feed utilization, high mortality, difficult motion and skin haemorrhage. Broken‐line regression analysis of AWG, FCR and PRE indicated that fingerling C. punctatus require niacin in the range of 37.1–42.1, whereas that of liver niacin concentration indicated the niacin requirement at 52.3 mg/kg dry diet.     

Growth performance and body composition in response to dietary protein and lipid levels in Nile tilapia (Oreochromis niloticus Linnaeus, 1758) subjected to normal and temporally restricted feeding regimes

A factorial experiment was designed to examine the effect on compensatory growth (CG) of Nile tilapia Oreochromis niloticus fed diets containing different protein and lipid levels under normal and temporally restricted feeding regimes. Four diets were formulated to contain either 30% or 36% crude protein, and 5% or 11% crude lipid. Triplicate replicates of each treatment were assigned to 24 150‐L tanks (20 fish/tank density). Fish (mean initial weight ± SD = 8.79 ± 0.34 g) were then fed either the normal feeding regime (thrice daily to apparent satiation) or the restricted regime (1 day feed deprivation followed by 3 days of feeding to apparent satiation) over a 44‐days study period. Fish receiving a diet under the restricted regime achieved weight gains (WG) comparable to fish consuming the diet containing 30% protein and 5% lipids under the normal feeding regime. Fish maintained on the restricted feeding regime exhibited reduced feed intake (FI), WG, feed efficiency ratio (FE), protein efficiency rate (PER) and hepatosomatic index versus fish on the normal feeding regime, except WG in fish fed the diet with 30% protein and 5% lipids. However, the resultant FI (85%~94%) was higher than the excepted 75% intake when fish were subjected to the restricted regime. Feeding 11% lipid diets led to improved FI, WG, FE, and PER compared to feeding the 5% lipid diets. Increased FI, WG, and FE, but reduced PER were observed in fish fed with 36% protein versus fish fed 30% protein. Fish receiving the 36% protein diets had lower whole‐body moisture and ash contents, but elevated whole‐body protein and lipid contents compared to those receiving the 30% protein diets. Whole‐body moisture contents were lower, but whole‐body protein, lipid and ash contents were higher in fish fed 11% lipid diets than in fish fed 5% lipid diets. There was an increase in whole‐body moisture content, but a decrease in protein and lipid content in response to the restricted feeding regime. Ash content was not affected by the feeding regime. The present study shows that Nile tilapia fed diets subjected to a restricted feeding regime exhibited growth comparable to those fed the diet at 30% protein and 5% lipid levels under a normal feeding regime. This positive effect was more pronounced in diets at a high protein level or in a combination of high protein and lipid levels.     

Marine ash‐products influence growth and feed utilization when Atlantic cod Gadus morhua L. are fed plant‐based diets

Atlantic cod Gadus morhua L. were fed high plant protein diets added to either shrimp‐shell meal or crab‐shell meal. The aims were to investigate if diluting dietary energy would reduce the liver index (HSI) and if marine ash would add value to plant protein‐based diets. Two control diets were used: a high plant protein control diet (PP) with no marine ash addition, and a fishmeal‐based diet (FM) with no marine ash addition. All diets were evaluated in small cod (initial weight 79 ± 15g) and in market‐size cod (initial weight 1579 ± 20 g). Addition of crab‐shell meal up to 20% and shrimp‐shell meal up to 10% did not influence liver size in either small or market‐size cod. An addition of up to 20% crab‐shell meal and 10% shrimp‐shell meal improved growth compared to the PP control diet, and stimulated increased feed intake. However, 10% shrimp‐shell meal and 20% crab‐shell meal diets resulted in a similar intake of energy and protein as the control groups. Increasing shrimp‐shell meal to 20% resulted in reductions in feed intake, fat digestibility and growth, and in altered gut histology. All diets, except the 20% shrimp added diet, resulted in normal ranges of plasma nutrients and blood hematological values, showing good fish health with or without the marine ash addition.     

Application of high-voltage electrical discharges and high-pressure homogenization for recovery of intracellular compounds from microalgae Parachlorella kessleri

Treatments with high-voltage electrical discharges (HVED) and high-pressure homogenization (HPH) were studied and compared for the release of ionic components, carbohydrates, proteins, and pigments from microalgae Parachlorella kessleri (P. kessleri). Suspensions (1% w/w) of microalgae were treated by HVED (40 kV/cm, 1–8 ms) or by HPH (400–1200 bar, 1–10 passes). Particle-size distribution (PSD) and microscopic analyses were used to detect the disruption and damage of cells. HVED were very effective for the extraction of ionic cell components and carbohydrates (421 mg/L after 8 ms of the treatment). However, HVED were ineffective for pigments and protein extraction. The concentration of proteins extracted by HVED was just 750 mg/L and did not exceed 15% of the total quantity of proteins. HPH permitted an effective release overall of intracellular compounds from P. kessleri microalgae including a large quantity of proteins, whose release (at 1200 bar) was 4.9 times higher than that obtained by HVED. Consequently, HVED can be used at the first step of the overall extraction process for the selective recovery of low-molecular-weight components. HPH can be then used at the second step for the recovery of remaining cell compounds.

     

Dietary inclusion of mussel meal enhances performance and improves feed and protein utilization in common sole (Solea solea,Linnaeus, 1758) juveniles

The present study was carried out to test different mussel meal (MM) dietary levels in combination with fishmeal (FM) on the growth performance, fatty acid composition and liver histology of common sole, Solea solea juveniles to highlight the growth potential of this species. Four isoproteic (53%) and isolipidic (11%) pelletized diets were formulated to contain graded levels of mussel meal, MM0 (0%), MM25 (25%), MM50 (50%) and MM75 (75%), up to 75%. Sole juveniles (initial individual mean body weight 13.1 ± 2.3 g, n = 840) were fed to satiation for 91 days. Seventy fish per tank (500‐L, 0.64 m² bottom surface) were reared in 12 tanks (3 tanks per treatment) at 20 ± 1°C. Diets containing MM (MM25, MM50 and MM75) gave a significantly higher specific growth rate (SGR, 1.27 ± 0.01, 1.38 ± 0.06 and 1.40 ± 0.05, respectively), higher feed intake and lower feed conversion rate (FCR, 1.09 ± 0.01, 1.00 ± 0.04 and 0.98 ± 0.02, respectively) when compared to the FM‐based diet (MM0, SGR, 0.98 ± 0.11, FCR, 1.52 ± 0.13). Carcass proximate composition was not influenced by dietary treatments, with the exception of the significantly lower lipid content in the MM75 group. Protein efficiency ratio (PER) and gross protein efficiency (GPE) were significantly improved by the mussel meal inclusion (PER, 1.29 ± 0.12, 1.76 ± 0.01, 1.89 ± 0.06, 1.95 ± 0.08; GPE, 25.29 ± 1.85, 33.38 ± 0.89, 35.96 ± 1.36, 36.59 ± 1.05 in MM0, MM25, MM50 and MM75, respectively). A significant decrease in the viscerosomatic index was observed in fish fed with MM50 and MM75 in comparison to MM0. The hepatosomatic index of fish fed with MM0 and MM25 was higher than that of fish fed with MM75, although the histological examination of liver parenchyma in all experimental groups showed a uniformly abundant accumulation of lipid droplets. Carcass fatty acid composition was significantly affected by dietary treatments, reflecting the dietary fatty acid profile. According to these results, the inclusion of MM in experimental FM‐based diets improved the performance and feed utilization of common sole juveniles. The inclusion of MM in the present trial allowed a higher SGR than that registered in previous growth trials on common sole. This study could provide useful information to detect effective ingredients for practical diets in Solea solea. It also seems advisable to consider an inclusion of at least 25% MM in the experimental reference diet to be used for further application towards the development of specific diets for this species.     

Two-stage recovery of S-adenosylmethionine using supported liquid membranes with strip dispersion

We report the selective recovery of S-adenosylmethionine (SAM) from fermentation broths using a two-stage supported liquid membrane system with strip dispersion (SLM-SD). The system utilized two MiniModule^® hollow-fiber membrane modules as microporous supports and an organic membrane solution consisting of the extractants of sodium di-2-ethylhexyl sulfosuccinate (AOT), di-(2-ethylhexyl)phosphoric acid (DEHPA), and trioctylphosphine oxide (TOPO) in the solvent n-octanol. SAM was extracted in the first membrane module. Methionine (Met) was captured by the first stripping solution and further purified in the second membrane module. pH values in the feed phase and the first and second stripping solutions, extractant concentrations, NaCl concentration, and the SAM acceptor in the first stripping solution were optimized. Strip dispersion mixing speed, pressure differences across the membranes, and flow rates of the feed and strip dispersion phases were investigated experimentally. The optimal extractant concentrations were: AOT 2.78 wt%, DEHPA 27.0 wt%, and TOPO 1.61 wt%. The optimal pH values in the feed phase and the first and second stripping solution were 3.0, 2.5, and 1.0, respectively. SAM extraction efficiency of 98.7%, SAM recovery efficiency of 91.8% and Met removal efficiency of 85.4% were achieved within 5 h. Finally, the mass transfer analysis indicated that the mass transfer resistances from the extraction reaction and the membrane phase were predominant.     

Effects of feeding rate on the growth performance of gynogenetic albino sterlet,Acipenser ruthenus (Linnaeus, 1758) larvae

The feeding rate effects were studied on the growth performance of gynogenetic diploid larvae of sterlet Acipenser ruthenus during the first 4 weeks of exogenous feeding. The experimental rearing was conducted from 7 to 38 days post‐hatch (dph) in a circulation system. This was set up in four groups with three replicates (440 individuals/replicate), viz: AC‐control larvae fed Artemia sp., CFC‐control larvae fed compound feed, AG‐gynogenetic larvae fed Artemia sp., and CFG‐gynogenetic larvae fed compound feed. The larvae were reared in glass tanks (44 L volume, 10 individuals/L) with the temperature maintained at 18 ± 0.5°C, photoperiod of 12L:12D and water flow regime of 1‐L/min and fed 50%, 25%, 25%, and 9% of their total biomass/day during feeding. Highest TL and WBW of gynogenetic diploid larvae (AG) were observed with 50.6 ± 1.2 mm and 607.3 ± 36.1 mg (n = 30) at 38 dph. Highest TL and WBW of control larvae (AC) were recorded with 49.5 ± 3.8 mm and 600.8 ± 88.0 mg (n = 30), respectively, with 73.1% ± 11.4% survival; the lowest survival rate was at 46.4% ± 7.1% (n = 30) for the CFG group. The results indicate that the gynogenetic and normal larvae of sterlet fed with live food (Artemia nauplii) from 7 dph can achieve higher growth and survivability compared to the larvae fed with formulated test feed. Results of this study suggest that the effective rearing of sterlet larvae from 7 to 38 dph strongly depends upon the types of feed rather than the genome manipulation performed.     

Characterization of epididymal sperm from Spix's yellow‐toothed cavies (Galea spixii Wagler, 1831) recovered by different methods

The aim of the present study was to characterize the epididymal sperm of Spix's yellow‐toothed cavy (Galea spixii) through two different recovery methods. Nine sexually mature males were euthanized and the complexes, testes–epididymis, were dissected. For each animal, one epididymis was processed by flotation method and the other was processed by retrograde flushing method, both using a TES‐based buffered media. After recovery, we evaluated the sperm for motility, vigour, viability, functional membrane integrity and morphology. Morphometric data from the different sperm regions were evaluated by using an appropriate software. After recovery, both methods provide similar values for all the sperm parameters, aiming the recovery of more than 300 × 10⁶ sperm, presenting >50% motile sperm, with normal morphology and functional membrane. The total sperm length in this sperm was 48.87 ± 0.1, and the sperm head presented 9.4%, on average. A notable characteristic was the prominent acrosome found in the G. spixii sperm. In conclusion, we demonstrate that either flotation or retrograde flushing methods are suitable for the recovery of sperm from cauda epididymis of Spix's yellow‐toothed cavies.     

Variability of in vitro ruminal fermentation and nutritional value of cell‐disrupted and nondisrupted microalgae for ruminants

The objective of this study was to investigate ruminal fermentation and the nutritional value of different microalgae products (MAP) for ruminants, including inter‐ and intra‐genera variability. Furthermore, the effect of mechanical cell disruption was also evaluated. Cell‐disrupted and nondisrupted MAP of four genera were investigated using the Hohenheim Gas Test. The investigations included characterization of gas production (GP), production of volatile fatty acids (VFA) and methane, organic matter digestibility, and energetic value as well as utilizable crude protein at the duodenum and ruminally undegradable crude protein (RUP). Furthermore, a three‐step enzymatic in vitro system was used to estimate intestinal digestibility of RUP (IDP). Ruminal fermentation was low for all investigated microalgae genera, as indicated by overall low GP, low production of VFA, and low ruminal protein degradation. Nevertheless, all microalgae genera were characterized by high RUP concentrations (236–407 g/kg dry matter; passage rate = 8% hr^?1), indicating that microalgae might be a promising protein source for high‐performing ruminants. Low IDP (26%–49% of RUP) considerably contradicted this potential. Mechanical cell disruption in general enhanced the extent of ruminal fermentation of MAP but, as RUP was decreased and IDP was hardly affected, mechanical cell disruption appears not to be necessary when microalgae are intended for application as a protein source for ruminants. Because of the high variability in the characteristics of the nutritional value, general means are inappropriate to characterize the nutritional value of MAP. In conclusion, suitability of microalgae as a protein source for ruminants might be limited because of low IDP, although further studies are necessary to prove these findings in vivo.     

Rapid harvesting of freshwater microalgae using chitosan

In this study, chitosan was used as a flocculant to harvest freshwater microalgae Chlorella vulgaris. The recovery efficiency of C. vulgaris was tested at various chitosan concentrations. 120 mg/L of chitosan showed the highest efficiency (92 ± 0.4%) within 3 min. The maximum concentration factor of 10 was also achieved at this dose of chitosan. The harvesting efficiency was pH dependent. pH 6.0 showed the highest harvesting efficiency (99 ± 0.5%). Measurement of zeta-potential confirmed that the flocculation was induced by charge neutralization. This study showed that a biopolymer, chitosan, can be a promising flocculant due to its high efficacy, low dose requirements, and short settling time.     

高温湿热酸法破壁提取法夫酵母胞内虾青素

法夫酵母是一种能积累虾青素的红酵母, 对其进行破壁是当前虾青素工业化提取生产的瓶颈工艺。研究在高温湿热条件下,低浓度盐酸对法夫酵母破壁而提取其胞内虾青素的工艺。探讨了不同破壁温度、盐酸浓度、液料比与破壁处理时间等因素对法夫酵母破壁后虾青素及类胡萝卜素提取率的影响, 确定了高温湿热酸法破壁提取虾青素的最佳条件为: 饱和蒸汽压力 0.1 MPa (121°C), 盐酸浓度0.5 mol/L, 液料比 (V/W)30 mL/g, 破壁时间2 min。在最佳条件下进行中试放大实验, 可得到虾青素与类胡萝卜素的提取率分别为: (84.8±3.2)%, (93.3±2)%。经优化后的新破壁工艺安全高效, 不会有毒性残留, 具有良好的工业应用前景。     

Effects of dietary amino acid l‐lysine on survival,growth and haemato‐biochemical parameters in Indian catfish,Heteropneustes fossilis (Bloch, 1974), fingerlings

An 8‐week feeding experiment was conducted to estimate the dietary lysine requirement of fingerling Indian catfish, Heteropneustes fossilis (total length = 6.35 ± 1.10 cm; weight = 4.70 ± 0.65 g) by feeding six isonitrogenous (400 g/kg CP) and isocaloric diets (17.90 kJ/g) containing six supplemental lysine levels ranging from 11.0 to 23.5 g/kg (diets I to VI, in incremental steps of 2.5 g/kg). Triplicate groups with 20 fish each were stocked in 75‐L circular continuous flow‐through troughs and fed experimental diets at 4% BW/day twice daily (08:00 and 18:00 h). Maximum live weight gain (304%), best‐feed conversion ratio (FCR, 1.46) and protein efficiency ratio (PER, 1.71) were obtained at 18.5 g/kg dietary lysine, beyond which these values showed a slight declining tendency. Best values for somatic and hematological indices were also obtained at this level. Significantly (p < .05) higher body protein and low carcass moisture and intermediate fat contents were noted in fish fed diets containing 18.5 g/kg lysine. The quadratic regression analyses of live weight gain, FCR, PER and body protein deposition (BPD) data indicated lysine requirements at 19.3, 18.8, 18.6 and 18.6 g/kg of dry diet, respectively. Based on these results, it is recommended that the H. fossilis diet should contain lysine at a level of 19.0 g/kg of dry diet, corresponding to 47.5 g/kg of dietary protein, for optimum growth and efficient feed utilization.     

Effect of papaya (Carica papaya) leaf extract as dietary growth promoter supplement in red hybrid tilapia (Oreochromis mossambicus × Oreochromis niloticus) diet

The purpose of this experiment was to examine the potential use of Carica papaya leaf extract as a supplement to promote growth and improve feed utilization in red hybrid tilapia. Five diets were formulated containing isolipidic (80 g/kg) and isonitrogenic (350 g/kg) levels. All feeds contained similar types and amounts of raw materials but differed in the inclusion of papaya leaf extract (0, 5, 10, 20 and 40 g/kg feed). The initial size of fish used was 2.3 ± 0.01 g. Each diet was performed in triplicate tanks, and the feeding period was 12 weeks. Fish fed diet containing 2% papaya leaf extract (PLE) had the highest final weight, 31.14 ± 1.47 g, followed by 1% PLE (27.27 ± 1.75 g). These two diets (1% and 2%) were also showed significant improvements of weight gain, SGR, and feed efficiency of the red hybrid tilapia (p < 0.05). However, papaya leaf extract did not affect the HSI, VSI, PER, digestive enzymes activity, blood composition, and survival rate. Supplementing the diets with papaya leaf extract lowered serum urea. Findings of this research suggest that adding papaya leaf extract to the diet of red hybrid tilapia improves growth and feed efficiency without adversely affecting blood parameters. Therefore, an inclusion level between 1% and 2% of the papaya leaf extract is recommended as a feed additive to promote red hybrid tilapia fry growth.     

High stocking density during larviculture and effect of size and diet on production of juvenile Lophiosilurus alexandri Steindachner, 1876 (Siluriformes: Pseudopimelodidae)

Different stocking densities were investigated in larviculture and feeding of Lophiosilurus alexandri, as well as analyses of the effects on juveniles of two size‐classes and two different commercial formulated diets. The first experiment was two‐phased: (a) larvae stocked at densities of 60, 120, 180, 240, and 300 larvae L⁻¹ fed with Artemia nauplii and reared for 15 days; (b) in phase 2, densities of 5, 10, 15, 30, and 40 juveniles L⁻¹ were evaluated during feed training (20 days). Mean water temperature in both phases was 28°C. In the first phase of experiment 1, the different stocking densities did not affect fish growth or survival. In phase 2, growth was similar in all densities; however, survival was lower at higher densities. The increased density provided a rise in biomass and number of individuals produced in both phases. In the second experiment, two size‐classes of feed‐trained juveniles (30.22 ± 1.84 and 34.66 ± 2.41 mm) were given pellets of two different diameters (1.2 and 2.6 mm) for 20 days. The largest juveniles fed the 1.2 mm inert diet had higher final weights and lengths. Larviculture and feed training of L. alexandri can thus be performed successfully at high stocking densities of 300 larvae L⁻¹ during the first 15 days of feeding, and at densities of up to 40 juveniles L⁻¹ during the 20 days of feed training, respectively.     

Evaluation of safflower meal as a protein source in diets of rainbow trout [Oncorhynchus mykiss,Walbaum, 1792)]*

The effects of different levels of safflower meal (a residue of the plant oil extraction process) in diets of rainbow trout (Oncorhynchus mykiss) on growth performance, nutrient digestibility and body composition were investigated. Four different diets containing 0% (Control group), 10% (Diet 1), 15% (Diet 2) and 20% (Diet 3) safflower meal were formulated for the fish with a mean weight of 132.70 ± 0.75 g fed for 70 days. At the end of the experiment, there were no significant differences among dietary treatments for weight gain, specific growth rate and feed conversion ratio (P > 0.05). There were also no differences in the digestibility of crude protein and crude lipid between groups (P > 0.05). Supplementation of safflower meal in the diets also had no adverse effects on body composition. The results indicate that safflower meal is a promising feed ingredient and can be used up to a concentration of 20% in the rainbow trout diet with no adverse effects on growth performance, nutrient digestibility or body composition.     

Development of a silicon carbide disruption method enables efficient extraction of proteins from cyanobacterium Synechocystis sp. PCC 6803

The oxygenic photosynthetic prokaryotes cyanobacteria have recently attracted worldwide interest in production of biofuels and bioactive natural compounds. Disruption of cells is a prerequisite for extraction of intracellular compounds. However, cyanobacterial cells are difficult to be disrupted because of the multiple-layered cell walls covered with a mucilaginous sheath. Here, we report a new disruption method for cyanobacteria, where an abrasive material, silicon carbide, is ground with cell pellet in situ. A cell disruption efficiency of 93.3 ± 2.3% was achieved in 6 min, an efficiency comparable to that obtained after 30 min of sonication. The new method yielded crude cell extracts with high concentrations of protein and high activity of the target enzyme, d-lactate dehydrogenase. This method has potential to be used for disruption of cells of photosynthetic microorganisms on a larger scale.     

Partial replacement of fishmeal in diets for turbot (Scophthalmus maximus,Linnaeus, 1758) culture using blue mussel (Mytilus edulis,Linneus, 1758) meat

The purpose of this study was to investigate whether meat from blue mussels (Mytilus edulis) harvested from offshore structures could be used as an alternative protein source in aquaculture fish feed for turbot (Scophthalmus maximus). Two feeding experiments with turbot were conducted in an initial experiment to test the applicability of mussel meal as a fishmeal replacement and/or supplement in turbot feed. In experiment A, the welfare, acceptance, and digestibility of feedstuff with 100%, 50% and 0% of mussel meal replacement for the fishmeal were tested for 8 weeks. In experiment B that followed, the convenience ratio of mussel meal protein in turbot feed for optimum growth was examined. Feedstuff with 25%, 10% and 0% of mussel meal were also fed for 8 (+6) weeks. Growth rates of fish were measured every 2 weeks and their health condition (liver somatic index) determined at the beginning and end of the experiments. Experiment A revealed that a replacement of fishmeal with either 100% or 50% mussel meal resulted in depressed growth (SGR: 1.31 ± 0.04 with 100% mussel meal; 1.54 ± 0.04 with 50% mussel meal; 1.71 ± 0.07 with 0% mussel meal). Experiment B revealed that a fishmeal replacement with 10% or 25% of mussel meal did not reduce growth nor did it have a negative effect on the health of the tested turbot. The study revealed that mussel meal has a high potential to serve as a supplement or as a fishmeal replacement in feed for turbot raised in aquaculture systems. Thus, its use in commercial aquaculture should be given future consideration.