Search Results (10)

Schizochytrium limacinum SR21, a kind of eukaryotic heterotrophic organism rich in unsaturated fatty acids, is an emerging microbial alternative to fish oil. The dietary inclusion of 15% SR21 was optimal for the growth performance of zebrafish. Previous studies demonstrated that fructose-1,6-bisphosphate aldolase (FBA) of Edwardsiella tarda is a valuable broad-spectrum antigen against various pathogens in aquaculture (e.g., Aeromonas hydrophila, Vibro anguillarum, Vibro harveyi, Vibro alginolyticus). We pioneered the development of stable S. limacinum SR21 transformants expressing the antigen protein FBA, exploring their potential as a novel oral vaccine for the aquaculture industry. The model animal zebrafish (Danio rerio) and ornamental fish koi carp (Cyprinus carpio koi) were harnessed to assess the immunoprotective effect, respectively. According to the quantitative expression analysis, zebrafish fed with recombinant Schizochytrium expressing FBA exhibited specific immune responses in the intestine. The expression levels of MHC-I and MHC-II, involved in cell-mediated adaptive immune responses, were significantly upregulated on the 14th and 28th days post-immunization. Additionally, the expression of highly specialized antibody genes IgZ1 and IgZ2 in mucosal immunity were significantly triggered on the 14th day post-immunization. Feeding koi carp with recombinant S. limacinum SR21-FBA increased the production of myeloperoxidase and FBA-specific antibodies in the sera. Furthermore, the sera of koi fed with recombinant S. limacinum SR21-FBA exhibited significant bactericidal activities against pathogen E. tarda. Thus, S. limacinum SR21 is a natural and highly promising oral vaccine carrier that not only provides essential nutrients as a functional feed ingredient but also offers specific immune protection to aquatic animals. This dual application is vital for promoting the sustainable development of the aquaculture industry. Full article

Hermetia illucens is a promising insect due to its ability to convert low-value substrates as food chain by-products into highly nutritious feed. Its feeding and nutrition are important issues. The aim of this work was to investigate the effect of different substrates consisting of coffee silverskin, a by-product of the roasting process, enriched with different inclusions of microalgae (5%, 10%, 20%, and 25%), Schizochytrium limacinum, and Isochrysis galbana, combined with the assessment of environmental sustainability by LCA. In general, the addition of microalgae led to an increase in larval growth performance due to the higher content of protein and lipids, although S. limacinum showed the best results with respect to larvae fed with coffee silverskin enriched with I. galbana. A higher prepupal weight was observed in larvae fed with 10%, 20%, and 25% S. limacinum; shorter development times in larvae fed with 25% of both S. limacinum and I. galbana; and a higher growth rate in larvae fed with 25% S. limacinum. The 10% S. limacinum inclusion was only slightly different from the higher inclusions. Furthermore, 10% of S. limacinum achieved the best waste reduction index. The greater the inclusion of microalgae, the greater the environmental impact of larval production. Therefore, the addition of 10% S. limacinum appears to be the best compromise for larval rearing, especially considering that a higher inclusion of microalgae did not yield additional benefits in terms of the nutritional value of H. illucens prepupae. Full article

Biodiesel produced from waste cooking oil (WCO) is on the rise and inevitably leads to issues in managing glycerol waste. Due to the presence of colour, odour and other minor compounds, the refining costs for this type of glycerol are higher and uneconomical. The potential of biodiesel-derived glycerol waste (BDGW) obtained from WCO to produce the highly added product of docosahexaenoic acid (DHA), also known as omega-3 polyunsaturated fatty acid, via the marine microalga of Schizochytrium limacinum ATCC MYA-1381 under aerobic batch fermentation was investigated. Cell growth, as well as DHA production, were performed under various operating conditions, including aeration rates and BDGW concentrations. The effect of the substrate type on cell growth and DHA yield was evaluated. The optimum operating condition was obtained when the air flow of a 0.25 vvm and 50 g/L of the glycerol concentration was fed into the fermenter and maximum cell dry weight (11.40 g/L) and DHA yield (665.52 mg/g) were achieved. However, cell growth and DHA yield were not significantly different when S. limacinum was grown using various carbon sources. Successfully, it clearly demonstrates that the BDGW can be used as a cheap carbon source for DHA production via marine microalgae using aerobic batch fermentation. Full article

The aim of the present study was to assess the effects of the low level of Schizochytrium limacinum marine algae (daily 5 g per animal) on the milk, cheese, and whey composition; fatty acid profile of milk and cheese; and the sensory profile of goat milk using an e-nose device. Thirty Alpine goats were randomly divided into two groups: the control group (C, n = 15)—fed grass with daily 600 g concentrate and the experimental group (MA, n = 15) who received the same forage and concentrate supplemented with 5 g/head/day marine algae. Animals were kept indoors and the investigation period lasted 52 days, including the first six weeks as the period of adaptation and the last 10 days as the treatment period. During the adaptation period, bulk milk samples from each group were collected once a week (0, 7, 14, 21, 28, 35, and 42 d), while during the treatment period (10 days), bulk milk samples from each group were taken every day, and cheese samples were processed from bulk milk each day from both groups. Marine algae supplementation had no negative effect on milk composition. In contrast, the marine algae inclusion significantly elevated the fat and protein content of whey and the protein content of cheese, as well as the recovery of fat and protein in the curd, while increasing the cheeses’ moisture content on a fat-free basis. The marine algae supplementation significantly increased the docosahexaenoic acid (DHA) and the rumenic acid (CLA c9t11) concentrations and decreased the n-6/n-3 ratio in the milk and cheese. There were no significant differences between the C and the MA group with regard to the sensory profiles of the milk. It can be concluded that the milk obtained from goats given daily supplementation of 5g of MA has a fatty acid profile more beneficial to human health, without any negative effects on the milk’s aromatic components. Full article

One of the most promising avenues of biofuel research relates to using waste as a starting feedstock to produce liquid or gaseous energy carriers. The global production of waste glycerol by the refinery industry is rising year after year. The aim of the present study was to examine the effect of ethyl methane sulfonate (EMS) on the growth rates and intracellular lipid accumulation in heterotrophically-cultured Schizochytrium limacinum microalgae, grown on waste glycerol as the carbon source. The strain S. limacinum E20, produced by incubating a reference strain in EMS for 20 min, was found to perform the best in terms of producing biomass (0.054 g_DW/dm³·h) and accumulating intracellular bio-oil (0.021 g/dm³·h). The selected parameters proved to be optimal for S. limacinum E20 biomass growth at the following values: temperature 27.3 °C, glycerol level 249.0 g/dm³, oxygen in the culture 26%, and yeast extract concentration 45.0 g/dm³. In turn, the optimal values for lipid production in an S. limacinum E20 culture were: temperature 24.2 °C, glycerol level 223.0 g/dm³, oxygen in the culture 10%, and yeast extract concentration 10.0 g/dm³. As the process conditions are different for biomass growth and for intracellular lipid accumulation, it is recommended to use a two-step culture process, which resulted in a lipid synthesis rate of 0.41 g/dm³·h. Full article

The ability of microalgae, such as the rich in docosahexaenoic acid (DHA) heterotrophic marine microalgae Schizochytrium limacinum SR21, to utilize nutrients in their culture media derived from low-cost nutrient sources makes them a promising low-cost alternative source for the production of useful substances used in aqua feeds. The assessment of culture parameters, one of which is the pH, for every different nutrient growth medium used for microalgae cultivation is important, as they affect the biomass and secondary metabolite microalgae production. This study assessed the effect of different growth medium pH levels (6, 7, 8 and 9), at laboratory and pilot scale systems, on S. limacinum biomass productivity, lipid accumulation, proximate composition, carbon assimilation and DHA. The microalgae were cultivated in growth media containing two different alternative low-cost nutrient sources: (a) crude glycerol derived from biofuel industry as carbon source and (b) effluent digestate from biogas production of livestock decomposition as a source of nutrients and trace elements. It was found that a neutral pH (7) was the optimum level, as it enhanced biomass productivity of the lab and pilot scale cultivation systems at 44.9 g L⁻¹ and 11 g L⁻¹ and DHA content at 7.5% and 19% of the total lipid content, respectively. Full article

The simultaneous use of crude glycerol and effluent from anaerobic digestate, both wastes derived from the biofuel industry, were tested in the frame of circular economy concept, as potential low-cost nutrient sources for the cultivation of rich in docosahexaenoic acid (DHA) oil microalgae strain Schizochytrium limacinum SR21. Initially, the optimal carbon and nitrogen concentration levels for high S. limacinum biomass and lipids production were determined, in a culture media containing conventional, high cost, organic nitrogen sources (yeast extract and peptone), micronutrients and crude glycerol at varying concentrations. Then, the effect of a culture media composed of crude glycerol (as carbon source) and effluent digestate at varying proportions on biomass productivity, lipid accumulation, proximate composition, carbon assimilation and fatty acid content were determined. It was shown that the biomass and total lipid content increased considerably with varying effluent concentrations reaching 49.2 g L⁻¹ at 48% (v/v) of effluent concentration, while the lipid yield at the same effluent concentration reached 10.15 g L⁻¹, compared to 17.0 g L⁻¹ dry biomass and 10.2 g L⁻¹ lipid yield when yeast extract and peptone medium with micronutrients was used. Compared to the control treatment, the above production was obtained with 48% less inorganic salts, which are needed for the preparation of the artificial sea water. It was shown that Schizochytrium limacinum SR21 was able to remediate 40% of the total organic carbon content of the biofuel wastes, while DHA productivity remained at low levels with saturated fatty acids comprising the main fraction of total fatty acid content. The results of the present study suggest that the simultaneous use of two waste streams from the biofuel industry can serve as potential nutrient sources for the growth of Schizochytrium limacinum SR21, replacing the high cost organic nutrients and up to one half the required artificial sea water salts, but upregulation of DHA productivity through optimization of the abiotic environment is necessary for industrial application, including aqua feed production. Full article

The polyketide synthase (PKS) cluster genes are supposed to synthesize polyunsaturated fatty acids (PUFAs) in S. limacinum. In this study, two enyolreductase (ER) genes located on PKS cluster were knocked out through homologous recombination to explore their functions. The knock-out of OrfB-ER (located on OrfB subunit) decreased lipid content and had obvious decrease on PUFAs content, indicating OrfB-ER domain played a vital role on PUFAs synthesis; the knock-out of OrfC-ER (located on OrfC subunit) decreased SFAs content and increased total lipid content, indicating OrfC-ER domain was likely to be related with SFAs synthesis, and lipid production could be improved by down-regulating OrfC-ER domain expression. Therefore, the addition of triclosan as a reported regulator of ER domain induced the increase of PUFAs production by 51.74% and lipids yield by 47.63%. Metabolic analysis indicated triclosan played its role through inhibiting the expression of OrfC-ER to reduce the feedback inhibition of SFAs and further to enhance NADPH synthesis for lipid production, and by weakening mevalonate pathway and tricarboxylic acid (TCA) cycle to shift precursors for lipid and PUFAs synthesis. This research illuminates functions of two ER domains in S. limacinum and provides a potential targets for improving lipid production. Full article

Schizochytrium limacinum residue was hydrolyzed with various proteases (papain, trypsin, Flavourzyme, Protamex, and Alcalase 2.4L) to obtain antioxidative peptides. The results showed that the S. limacinum hydrolysates (SLHs) prepared with compound proteases (Protamex and Alcalase 2.4L) had the highest antioxidant activity, which was measured using methods such as 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging ability (IC₅₀ = 1.28 mg/mL), hydroxyl radical scavenging ability (IC₅₀ = 1.66 mg/mL), and reducing power (1.42 at 5.0 mg/mL). The hydrolysates were isolated and purified by ultrafiltration, gel filtration chromatography, and reverse-phase high-performance liquid chromatography (RP-HPLC). Through analysis of electrospray ionization-mass spectrometer (ESI-MS/MS), the purified antioxidant peptide was identified as Pro-Tyr-Lys (406 Da). Finally, the identified peptide was synthesized for evaluating its antioxidant activity. The •OH scavenging ability and reducing power of Pro-Tyr-Lys were comparable to those of reduced L-glutathione (GSH). These results demonstrated that the antioxidant peptides from SLHs could potentially be used as effective antioxidants. Full article

The Cre/loxP site-specific recombination system was applied to Aurantiochytrium limacinum to obtain a transformant without the antibiotic resistance marker gene. First, the enhanced green fluorescent protein gene (egfp) and chloramphenicol resistance gene (Cm^r), along with the two loxP loci, were integrated into the genome of A. limacinum OUC88 using 18S rDNA sequences as the homologous recombination sites. Then plasmid pSH65, containing a zeocin resistance gene (Ble^r) was transferred into A. limacinum OUC_CG. After induction with galactose, repeated passage in culture and PCR-based assessment, the pSH65 plasmid was lost and A. limacinum OUC_EG host was shown to no longer have resistance to 100 mg chloramphenicol/L or 5 mg zeocin/L. Through southern blotting and fluorescence detection, egfp was found to be integrated into the genome of A. limacinum OUC_EG, and EGFP was successfully expressed in the cells. The successful application of the Cre/loxP system demonstrates an experimental basis for genetic modification of A. limacinum so as to obtain transformed strains with no antibiotic resistance marker genes. Full article