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Microalgae are considered to be natural producers of bioactive pigments, with the production of pigments from microalgae being a sustainable and economical strategy that promises to alleviate growing demand. Chlorophyll, as the main pigment of photosynthesis, has been widely studied, but its medicinal applications as an antioxidant, antibacterial, and antitumor reagent are still poorly understood. Chlorophyll is the most important pigment in plants and algae, which not only provides food for organisms throughout the biosphere, but also plays an important role in a variety of human and man-made applications. The biological activity of chlorophyll is closely related to its chemical structure; its specific structure offers the possibility for its medicinal applications. This paper reviews the structural and functional roles of microalgal chlorophylls, commonly used extraction methods, and recent advances in medicine, to provide a theoretical basis for the standardization and commercial production and application of chlorophylls.

Some mixotrophic microalgae appear to exceed the sum of photoautotrophy and heterotrophy in terms of biomass production. This paper mainly reviews the carbon and energy metabolism of microalgae to reveal the synergistic mechanisms of the mixotrophic mode from multiple aspects. It explains the shortcomings of photoautotrophic and heterotrophic growth, highlighting that the mixotrophic mode is not simply the sum of photoautotrophy and heterotrophy. Specifically, microalgae in mixotrophic mode can be divided into separate parts of photoautotrophic and heterotrophic cultures, and the synergistic parts of photoautotrophic culture enhance aerobic respiration and heterotrophic culture enhance the Calvin cycle. Additionally, this review argues that current deficiencies in mixotrophic culture can be improved by uncovering the synergistic mechanism of the mixotrophic mode, aiming to increase biomass growth and improve quality. This approach will enable the full utilization of advantagesin various fields, and provide research directions for future microalgal culture., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Phytoplankton composition is an important factor affecting the growth and physiological biochemical characteristics of filter-feeding bivalves. With the increasing trend in dinoflagellate biomass and blooms in mariculture areas, how the physio-biochemical traits and seafood quality of the mariculture organism are affected by the dinoflagellates, especially those at nonfatal levels, is not well understood. Different densities of two Karlodinium species, namely K. veneficum (KV) and K. zhouanum (KZ), mixed with high quality microalgal food Isochrysis galbana was applied in feeding manila clam Ruditapes philippinarum in a 14-day temporary culture, to comparatively study how the critical biochemical metabolites such as glycogen, free amino acids (FAAs), fatty acids (FAs), volatile organic compounds (VOCs) in the clam were affected. The survival rate of the clam showed dinoflagellate density and species specificity. The high-density KV group inhibited survival to 32% lower than that of the pure I. galbana control, respectively, while KZ at low concentrations did not significantly affect the survival compared with the control. In the high-density KV group, the glycogen and FAA contents decreased (p < 0.05), indicating that energy and protein metabolism were significantly affected. Amount of carnosine (49.91 ± 14.64 to 84.74 ± 8.59 μg/g of muscle wet weight) was detected in all the dinoflagellate-mixed groups, while it was not present in the field samples or in the pure I. galbana control, showing that carnosine participated in the anti-stress activities when the clam was exposed to the dinoflagellates. The global composition of FAs did not significantly vary among the groups. However, contents of the endogenous C 18 PUFA precursors linoleic acid and α-linolenic acid significantly decreased in the high-density KV group compared to all the other groups, indicating that high density of KV affected the metabolisms of fatty acids. From the results of the changed VOC composition, oxidation of fatty acids and degradation of free amino acids might occur in the clams exposed to dinoflagellates. The increased VOCs, such as aldehydes, and decreased 1-octen-3-ol probably produced a more fishy taste and reduced food flavor quality when the clam was exposed to the dinoflagellates. This present study demonstrated that the biochemical metabolism and seafood qulity of the clam were affected. However, KZ with moderate density in the feed seemed to be beneficial in aquaculture for increasing the content of carnosine, a high-valued substance with multiple bioactivities., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Microalgae cannot meet the bait demand for aquaculture due to light intensity limitation and other disadvantageous conditions. This research selected 6 mixotrophic microalgae, and the optimal strains and organic carbon were screened. The results showed that Thalassiosira pseudonana and Chlorella sp. are suitable for mixotrophic culture. The maximum cell density of Thalassiosira pseudonana was found to be 1.67 times than that of the photoautotrophic group when glycerol was added. The maximum cell density of Chlorella sp. with acetic acid was 1.69 times than that of the photoautotrophic group. When the concentration of acetic acid was 5.0 g·L -1 and the concentration of KNO 3 was 0.2 g·L -1 , the maximum biomass of Chlorella sp. could reach 3.54 × 10 7 cells·mL -1 ; the maximum biomass of Thalassiosira pseudonana was 5.53 × 10 6 cells·mL -1 with 10.0 g·L -1 glycerol and 0.2 g·L -1 KNO 3 . Metabolomic analysis further revealed that mixotrophic bait microalgae could promote the accumulation of lipids and amino acids., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

The exploitation of new economically valuable microalgae as a sustainable source of minor high-value products can effectively promote the full utilization of microalgae. The efficient preparation of minor products from microalgae remains the challenge, owing to the coexistence of various components with a similar polarity in the microalgae biomass. In this study, a novel approach based on the sustainable-oriented strategy for fucoxanthin (FX) production was proposed, which consisted of four steps, including the culture of microalga, ethanol extraction, ODS column chromatography, and ethanol precipitation. The high-purity FX (around 95%) was efficiently obtained in a total recovery efficiency of 84.28 ± 2.56%. This study reveals that I. zhangjiangensis is a potentially promising feedstock for FX production and firstly provides a potentially eco-friendly method for the scale-up preparation of FX from the microalga I. zhangjiangensis ., Competing Interests: The authors declare no conflict of interest.

Marine microalgae offer a promising feedstock for biofuels and other valuable compounds for biorefining and carry immense potential to contribute to a clean energy and environment future. However, it is currently not economically feasible to use marine algae to produce biofuels, and the potential bioactive chemicals account for only a small market share. The production of algal biomass with multiple valuable chemicals is closely related to the algal species, cultivation conditions, culture systems, and production modes. Thus, higher requirements for screening of dominant algal strains, developing integrated technologies with the optimum culture conditions, efficient cultivation systems, and production modes to exploit algal biomass for biorefinery applications, are all needed. This review summarizes the screening of dominant microalgae, discusses the environmental conditions that may affect the growth, as well as the culture systems and production modes, and further emphasizes the valorization options of the algal biomass, which should help to offer a sustainable approach to run a profitable marine algae production system., Competing Interests: Declaration of competing interest The authors declare that we do not have any commercial or associative interest that represents a conflict of interest in connection with the work submitted., (Published by Elsevier B.V.)

Furfural wastewater (FWW) treatment is important in modern chemical production. However, traditional wastewater treatment methods are inappropriate for the treatment of FWW. In this work, Chlorella pyrenoidosa was employed to study the growth and pollutants removal with both heterotrophic and mixotrophic cultures. The results show that the biomass and removal efficiency for COD and TN were the highest under 10-fold dilution. However, TP removal were inconsistent when the algae were cultivated in both mixotrophic and heterotrophic modes. Compared to high nitrogen (0.75 g/L NaNO 3 ), the algal cells grew faster when adding 0.25 g/L NaNO 3 to the FWW, whether in mixotrophic or heterotrophic conditions. The total lipid content in heterotrophic conditions was 18.53%, which was higher than the values in mixotrophy when the concentration of NaNO 3 was 0.75 g/L. Different carbon assimilation mechanisms of the algal cells result in a discrepancy in cell growth and pollutant removal, under different culture modes., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

This research of mixotrophic microalgae Isochrysis 3011 with glycerol was combined with the treatment of aqua-cultural wastewater, different initial concentrations, and optimized light intensities. The algae growth rate, removal efficiencies of total nitrogen (TN) and total phosphorus (TP) were determined. Results showed that the suitable initial concentration was 0.4 g L -1 , and the optimum light intensity was 60 µmol m -2  s -1 . The growth of the mixotrophic group was better than that of the autotrophic culture. The biomass yield of the mixotrophic group with glycerol was 0.17 g L -1 d -1 , and the removal rates of TN and TP were 73.39% and 95.61%, respectively. The content of total lipid and total protein in mixotrophic group were higher than the values of the autotrophic group. This indicates that aquaculture wastewater treatment with mixotrophic bait microalgae can obtain superior micro-algal biomass, which is also a potential technology for wastewater utilization and ecological protection., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Phytohormones are a class of small organic molecules that are widely used in higher plants and microalgae as chemical messengers. Phytohormones play a regulatory role in the physiological metabolism of cells, including promoting cell division, increasing stress tolerance, and improving photosynthetic efficiency, and thereby increasing biomass, oil, chlorophyll, and protein content. However, traditional abiotic stress methods for inducing the accumulation of energy storage substances in microalgae, such as high light intensity, high salinity, and heavy metals, will affect the growth of microalgae and will ultimately limit the efficient accumulation of energy storage substances. Therefore, the addition of phytohormones not only helps to reduce production costs but also improves the efficiency of biofuel utilization. However, accurate and sensitive phytohormones determination and analytical methods are the basis for plant hormone research. In this study, the characteristics of phytohormones in microalgae and research progress for regulating the accumulation of energy storage substances in microalgae by exogenous phytohormones, combined with abiotic stress conditions at home and abroad, are summarized. The possible metabolic mechanism of phytohormones in microalgae is discussed, and possible future research directions are put forward, which provide a theoretical basis for the application of phytohormones in microalgae.

This research 12 microalgal species were screened for biofilm attached culture in the treatment of anaerobically digested swine wastewater (ADSW). The influence of ADSW on biomass productivity and removal efficiencies were evaluated using biofilm attached culture with the selected Chlorella pyrenoidosa. The variation of nutritional components from algal cells were further analysed to evaluate the potential applications of C. pyrenoidosa. The results showed that C. pyrenoidosa had the highest tolerance to ADSW, and the highest removal efficiencies for wastewater pollutants were reached when cultured in 5 times diluted ADSW. These test conditions resulted in an algal cell biomass composed of 57.30% proteins, 14.87% extracellular polysaccharide, 3.08% crude fibre, 5.57% crude ash, 2.85% moisture. Amino acids in proteins contained 21.73% essential amino acids and the EAA/NEAA value was 0.64. The essential amino acid score indicates that the selected C. pyrenoidosa could be a good protein source for feed addition., (Published by Elsevier Ltd.)

Cultivating microalgae on wastewaters is an effective way to produce algal biomass whereas harvesting microalgae is a costly operation. This study we examined the feasibility of co-culturing a high-value microalga with an auto-flocculating strain to enable efficient recovery of biomass. Experiments were conducted to co-cultivate Chlorella zofingiensis with Tribonema sp. on swine wastewater diluted by fishery wastewater under different conditions. The result showed the optimal inoculum ratio of Tribonema sp. to Chlorella zofingiensis was 1:1. The removal efficiencies of pollutants (NH 4 + -N, TN, TP, and COD) and lipid content were high when the co-culture ratios of Tribonema sp. were high. Also, some larger chain fatty acids, specifically C20:5 and C22:6 were present when the two strains co-culture. The recovery efficiency increased with the increasing proportion of auto-flocculating Tribonema sp.. Algae co-culture has the potential to address limitations in substrate utilization by individual strains, also improve the recovery of biomass., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Botryococcus braunii is known for its high yield of extracellular hydrocarbons and polysaccharides. Hydrocarbons, especially botryococcenes and squalene can be used as not only fuels but also alternative feedstock for other fossil-based products. Exopolysaccharides excreted from B. braunii can be used as scaffolds for polyesters production, and have a notable potential for synthesis of nanoparticles. B. braunii is also a rich source of carotenoids, especially the unique secondary carotenoids such as botryoxanthins that have never been found in other microalgae. The morphology, physiology, and outer cell walls of B. braunii are complex. Understanding the colony structure shall provide insights into the mechanism of cell growth and chemicals secretion. It is possible to improve the production economics of the alga with advanced culture systems. Moreover, investigation of metabolic pathways for B. braunii may help us understand their regulation and provide valuable information for strain selection and optimal production of high-value chemicals., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Background: Steryl glycosides (SGs) are sterol conjugates found in various plants, especially in those making up human diets. It has been demonstrated that SGs have potential health benefits, and they could be used as food supplements in a variety of food matrixes. Marine microalgae are a potential resource for human food and ingredients. In this study, gas chromatography-triple quadrupole mass spectrometry (GC-QQQ-MS) was used to characterize unknown SGs in eight microalgae belonging to different classes (Isochrysis galbana 3011, Pavlova viridis, Platymonas helgolandica, Conticribra weissflogii, Thalassiosira pseudonana, Nitzschia closterium, Gymnodinium sp., and Karlodinum veneficum)., Results: The SGs were first extracted from lyophilized algae with chloroform-methanol, purified by solid-phase extraction and analyzed as trimethylsilyl derivatives. Nine SGs have been identified. In particular, new SGs like occelasteryl glycoside and stellasteryl glycoside were found in Gymnodinium sp., 24-methylene cholesteryl glycoside was detected in P. helgolandica, and 4,24-dimethylcholestan-3-yl glycoside was identified as the main constituent of microalga K. veneficum. The results also showed that the compositions of SGs in different microalgae varied, with a range of 5.234 to 0.036 g kg -1 , and microalga P. viridis contained the most abundant SGs., Conclusion: GC-QQQ-MS is a powerful tool to detect SGs with different structures from a variety of microalgae. The compositions of SGs in different microalgae varied greatly. Microalgae are a good source of highly valued SGs. © 2017 Society of Chemical Industry., (© 2017 Society of Chemical Industry.)

Sphingolipid compositions are crucial for the structural and physiological properties of microalgae membranes. In the present study, we developed a quadrupole time-of-flight (Q-TOF) mass spectrometric method based on MS E data collection for the identification of sphingolipids with high efficiency, selectivity, sensitivity and mass accuracy and applied this method for precise structural identification and quantitative profiling of ceramides and glycosphingolipids in total lipid extracts from 17 strains of microalgae, including 11 strains of diatom, 3 strains of dinoflagellate and 3 strains of haptophyta. Using this method, four species of sphingolipids including 27 ceramides, 13 monosaccharide ceramides, 18 disaccharide ceramides and 18 trisaccharide ceramides were identified. The compositions of sphingolipid-included glycosyl moieties, long chain bases and N-acyl chains showed a significant difference among different microalgae categories. Some long chain bases including d19:2, d19:3 and d19:4, glycosyl moieties including disaccharide and trisaccharide, and N-acyl chains such as 14:0, 14:1, 24:0, 24:1, h18:1, h19:1 and h22:0-2 can be chosen as the molecular signature for microalgae from three major phyla. This methodology will be useful for a wide range of physiological and pathological studies of sphingolipids. Furthermore, the diversity of sphingolipid structure could provide a new criterion for microalgae chemotaxonomy., (Copyright © 2017. Published by Elsevier B.V.)

Characterization of the feeding preferences of bivalve larvae would help improving the bivalve aquaculture and hatchery by providing appropriate microalgal diets. However, inaccurate and laborious identification and counting of microalgal species have challenged the selective feeding of bivalves. In the present study, we developed a highly specific and sensitive assay using quantitative polymerase chain reaction (qPCR) to assess the selective feeding of bivalve larvae based on species-specific primers targeting to microalgal 18S rDNA sequences. The assay exhibited good specificity. The detection limits of the qPCR assay were 769, 71, 781 and 21 18S rDNA copies for Chaetoceros calcitrans, Isochrysis galbana, Platymonas helgolandica and Nannochloropsis oculata, respectively. Using such assay, we found that C. calcitrans and I. galbana were preferentially ingested, whereas N. oculata was preferentially rejected in biodeposits of four bivalve species, Tegillarca gransa, Cyclina sinensis, Scapharca subcrenata and Sinonovacula constricta. Furthermore, our growth experiments revealed that C. calcitrans and I. galbana could significantly promote the shell growth, whereas feeding of N. oculata resulted in poorer growth of four bivalve species. These data indicated that qPCR might be useful in screening of efficient and reliable microalgal species for each bivalve species, leading to improved bivalve aquaculture and hatchery.

Background: Head space solid-phase microextraction-gas chromatography-mass spectrometry has been applied to analyze the volatile components of six marine microalgae (Thalassiosira weissflogii, Nitzschia closterium, Chaetoceros calcitrans, Platymonas helgolandica, Nannochloropsis spp. and Dicrateria inornata) from Bacillariophyta, Chlorophyta and Chrysophyta, respectively, in different growth phases., Results: All volatile compounds were identified by database searching in the NIST08 Mass Spectral Library and analyzed by principal component analysis with SIMCA-P software (Umetrics, Umea, Sweden). The results clearly revealed that the volatile components of the six microalgae were significantly different in the exponential, stationary and declining phases. Aldehydes, alkanes, some esters and dimethyl sulfide significantly changed in different growth phases., Conclusion: This is the first report on the comprehensive characteristics of volatile components in different microalgae and in different growth phases. The results may provide reference data for studies on the flavor of cultivated aquatic organism, odor formation in nature water, choice of feeding period and microalgae species selection for the artificial rearing of marine organisms. © 2016 Society of Chemical Industry., (© 2016 Society of Chemical Industry.)

Rationale: Glycosphingolipids (GSLs) are important components of the cell membrane; however, little is known about GSLs in microalgae. We analyzed GSLs in three strains of Skeletonema microalgae by liquid chromatography coupled with mass spectrometry., Methods: Total lipid was extracted from Skeletonema microalgae. Separation of lipids was achieved via reversed-phase liquid chromatography, and mass spectrometric analysis of the lithium adduct of GSLs was conducted to determine their structures., Results: Two types of novel glycosphingolipids were identified from three strains of Skeletonema microalgae. The N-acyl groups were primarily long-chain saturated or monoenoic fatty acids (monounsaturated fatty acids) with 16C and 20 to 24C. The sphingoid long-chain bases (LCB) were sphingosine, sphingadienine and sphingatrienine. The saccharide polar head groups of glycosphingolipids were disaccharide (heptose-hexose) or trisaccharide (heptose-hexose-hexose). Semi-quantitative analysis of GLSs in the three strains of microalgae showed that GSL contents ranged from 0.09 to 8.79 nmol/mg dry microalgae powder., Conclusions: A qualitative method was developed for the identification of GSLs in microalgae. Two types of novel GSLs were identified from three strains of Skeletonema, which might have important biological functions. It could also provide a reliable tool for chemotaxonomy of microalgae., (Copyright © 2013 John Wiley & Sons, Ltd.)

A facile method based on liquid chromatography coupled with electrospray ionization tandem triple quadrupole mass spectrometry working in selected reaction monitoring mode has been established to analyze toxins in the algae and water samples. Twelve types of toxins (anatoxin, cylindrospermopsin, dinophysistoxin-1, nodularin, okadaic acid, microcystins) were efficiently separated under optimized liquid chromatography coupled with mass spectrometry conditions in the selected reaction monitoring mode. Correlation coefficients of the calibration curves, all felt in the range of 0.9958-0.9998, indicated good linearity. The detection limits of toxins in this method were all lower than 0.20 ng/mL and the quantification limits were in the range from 0.04 to 0.60 ng/mL. Except for anatoxin, cylindrospermopsin, and nodularin, the other toxins' recoveries varied from 55.45 to 140.85%. And the relative standard deviations of interday and intraday precision were at 8.61% (n = 5). The high-performance liquid chromatography (HPLC)/electrospray ionization (ESI)-mass spectrometery (MS) method was also successfully applied to analyze the algae and water samples. Owing to its exclusive selectivity and excellent sensitivity, the developed method is a tool for comprehensive analyses of the 12 types of toxins at nanogram levels., (© 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

This study has investigated the effects of six different unialgal diets ( Chaetoceros calcitrans , Platymonas helgolandica , Chlorella sp., Isochrysis galbana , Nannochloropsis oculata , and Pavlova viridis ) on the composition of fatty acids and sterols in juvenile ark shell Tegillarca granosa Linnaeus. The best feeding effects on the growth of shellfish were found in C. calcitrans, followed by I. galbana and P. viridis, whereas Chlorella sp. and N. oculata exhibited relatively poor effects. The fatty acid and sterol compositions in the six microalgae and the juvenile ark shell after feeding were analyzed, and 39 fatty acids and 18 sterols were identified. Although the results demonstrate a close correlation between the sterol compositions in algal species and juvenile ark shell, a similar correlation was not observed between fatty acids. In the juvenile ark shell fed microalgae, the ratio of total saturated fatty acids (SFA) rapidly decreases, whereas the proportion of total polyunsaturated fatty acids (PUFAs) increases considerably. The abundances of AA, EPA, and DHA increase most significantly in shellfish with better growth (fed C. calcitrans, I. galbana, and P. viridis). The number of sterol species is reduced, but the total sterol content in groups fed corresponding microalgae increases, and abundant plant sterols, instead of cholesterol, are accumulated in juvenile ark shell fed appropriate microalgae I. galbana and P. viridis. Therefore, to be more conducive to human health, I. galbana and P. viridis, of the six experimental microalgae, are recommended for artificial ark shell culture.

Abstract: BACKGROUND: Steryl glycosides (SGs) are sterol conjugates found in various plants, especially in those making up human diets. It has been demonstrated that SGs have potential health benefits, and they could be used as food supplements in a variety of food matrixes. Marine microalgae are a potential resource for human food and ingredients. In this study, gas chromatography–triple quadrupole mass spectrometry (GC–QQQ‐MS) was used to characterize unknown SGs in eight microalgae belonging to different classes (Isochrysis galbana 3011, Pavlova viridis, Platymonas helgolandica, Conticribra weissflogii, Thalassiosira pseudonana, Nitzschia closterium, Gymnodinium sp., and Karlodinum veneficum). RESULTS: The SGs were first extracted from lyophilized algae with chloroform–methanol, purified by solid‐phase extraction and analyzed as trimethylsilyl derivatives. Nine SGs have been identified. In particular, new SGs like occelasteryl glycoside and stellasteryl glycoside were found in Gymnodinium sp., 24‐methylene cholesteryl glycoside was detected in P. helgolandica, and 4,24‐dimethylcholestan‐3‐yl glycoside was identified as the main constituent of microalga K. veneficum. The results also showed that the compositions of SGs in different microalgae varied, with a range of 5.234 to 0.036 g kg−1, and microalga P. viridis contained the most abundant SGs. CONCLUSION: GC–QQQ‐MS is a powerful tool to detect SGs with different structures from a variety of microalgae. The compositions of SGs in different microalgae varied greatly. Microalgae are a good source of highly valued SGs. © 2017 Society of Chemical Industry [ABSTRACT FROM AUTHOR]

Rationale Diacylgycerol- N-trimethylhomoserine (DGTS) and diacylglycerylhydroxymethyl- N,N,N-trimethyl-β-alanine (DGTA) are structural isomers that are the most commonly described betaine lipids in microalgae. The structural differentiation and precise identification of DGTS and DGTA in microalgae need to be established during mass spectrometry analysis. Methods Total lipid was extracted from Amphora spp. with CHCl3/CH3OH (1:1, v/v). The qualitative analysis of DGTS and DGTA in Amphora spp. was carried out using Li+/H+ dual mode by ultra-performance liquid chromatography/quadrupole time-of-flight mass spectrometry operating in MSE mode (UPLC/QTOF MSE). Results Characteristic fragment ions [C10H22O5N]+ at m/z 236.15 and [C7H14O2N]+ at m/z 144.10 from the [M + H]+ precursor ion can be used for the qualitative analysis of both DGTA and DGTS, whereas the loss of m/z 87 and 74 from the [M + Li]+ precursor ion are specific for DGTS, and the loss of m/z 103 from the [M + Li]+ precursor ion is only for DGTA. As a result, 9 DGTSs and 16 DGTAs with different fatty acids were identified simultaneously in Amphora spp. Semi-quantitative analysis of DGTS and DGTA in Amphora spp. showed that the contents of DGTS ranged from 0.003 to 0.438 nmol mg-1 dw, and that of DGTA from 0.004 to 0.414 nmol mg-1 dw. Conclusions This is the first report to achieve the ambiguous structural identification of DGTS and DGTA by UPLC/QTOF MSE using dual Li+/H+ adduct ion mode, which has remained a challenge in the past. It could provide new insights into their phylogeny and be helpful to characterize the natural phytoplankton communities as intact polar lipid biomarkers. Copyright © 2017 John Wiley & Sons, Ltd. [ABSTRACT FROM AUTHOR]

Isochrysis galbana is a species of marine microalgae that is principally used to feed the larvae and early juveniles of bivalve mollusks (e.g., ark shells, clams, mussels, oysters and scallops). The artificial hatchery stages of most of these larvae are primarily occur during high-temperature seasons, when the water temperature in the breeding pool frequently exceeds 30 °C and even reaches 35 °C, which may change the lipid composition of I. galbana, thus affecting its nutritional value. In this experiment, we investigated changes in the lipid composition of I. galbana Parke at high temperature using ultra-performance liquid chromatography electrospray ionization quadrupole time-of-flight mass spectrometry. The results showed that the sulfoquinovosyldiacylglycerol (SQDG), monogalactosyldiacylglycerol (MGDG), diacylglycerylcarboxyhydroxymethylcholine (DGCC) and digalactosyldiacylglycerol (DGDG) contents increased during growth at 20 and 35 °C. Conversely, the lyso-SQDG, lyso-MGDG, lyso-DGDG, lyso-DGCC and triacylglycerol (TAG) contents clearly decreased. Moreover, higher SQDG, MGDG, DGDG and DGCC contents and lower lyso-lipid and TAG contents were observed in I. galbana Parke grown at 35 versus 20 °C. In addition, the content of docosahexaenoic acid (DHA) improved at 20 and 35 °C during growth, and DHA accumulated to greater levels at 35 °C than at 20 °C at the same time points. These results revealed an increase in DHA content as the temperature rose, which indicated that the nutritional value of I. galbana Parke during logarithmic phase improved in the same cultured time with an increasing temperature. [ABSTRACT FROM AUTHOR]

Rapid, high-throughput and reliable methods are urgently required to accurately detect and monitor harmful algae, which are responsible for algal blooms, such as red and green tides. In this study, we successfully developed a multiplex PCR-based DNA microarray method capable of detecting nine harmful algal species simultaneously, namely Alexandrium tamarense, Gyrodinium instriatum, Heterosigma akashiwo, Karenia mikimotoi, Prorocentrum donghaiense, Prorocentrum minimum, Ulva compressa, Ulva ohnoi and Ulva prolifera. This method achieved a limit of detection (LOD) of 0.5 ng of genomic DNA (orders of magnitude of the deci-nanogram range) in the tested algae cultures. Altogether, 230 field samples from ship ballast waters and seaport waters were used to evaluate the DNA microarray. The clinical sensitivity and specificity of the DNA microarray assay in detecting field samples were 96.4% and 90.9%, respectively, relative to conventional morphological methods. This indicated that this high-throughput, automatic, and specific method is well suited for the detection of algae in water samples. [ABSTRACT FROM AUTHOR]

The two morphologically similar microalgae NMBluh014 and NMBluh-X belong to two different strains of Nannochloropsis oceanica. They possess obviously different feeding effects on bivalves, but are indistinguishable by 18S rRNA and morphological features. In this work, lipidomic analysis followed by principal component analysis and orthogonal projections to latent structures discriminant analysis provided a clear distinction between these strains. Metabolites that definitively contribute to the classification were selected as potential biomarkers. The most important difference in polar lipids were sulfoquinovosyldiacylglycerol (containing 18:1/16:0 and 18:3/16:0) and monogalactosyldiacylglycerol (containing 18:3/16:3 and 20:5/14:0), which were detected only in NMBluh-X. Additionally, an exhaustive qualitative and quantitative profiling of the neutral lipid triacylglycerol ( TAG) in the two strains was carried out. The predominant species of TAG containing 16:1/16:1/16:1 acyl groups was detected only in NMBluh-X with a content of ~93.67 ± 11.85 nmol · mg−1 dry algae at the onset of stationary phase. Meanwhile, TAG containing 16:0/16:0/16:0 was the main TAG in NMBluh014 with a content of 40.25 ± 3.92 nmol · mg−1. These results provided the most straightforward evidence for differentiating the two species. The metabolomic profiling indicated that NMBluh-X underwent significant chemical and physiological changes during the growth process, whereas NMBluh014 did not show such noticeable time-dependent metabolite change. This study is the first using Ultra Performance Liquid Chromatography coupled with Electrospray ionization-Quadrupole-Time of Flight Mass Spectrometry (UPLC-Q-TOF-MS) for lipidomic profiling with multivariate statistical analysis to explore lipidomic differences of plesiomorphous microalgae. Our results demonstrate that lipidomic profiling is a valid chemotaxonomic tool in the study of microalgal systematics. [ABSTRACT FROM AUTHOR]

Phytohormones are a class of small organic molecules that are widely used in higher plants and microalgae as chemical messengers. Phytohormones play a regulatory role in the physiological metabolism of cells, including promoting cell division, increasing stress tolerance, and improving photosynthetic efficiency, and thereby increasing biomass, oil, chlorophyll, and protein content. However, traditional abiotic stress methods for inducing the accumulation of energy storage substances in microalgae, such as high light intensity, high salinity, and heavy metals, will affect the growth of microalgae and will ultimately limit the efficient accumulation of energy storage substances. Therefore, the addition of phytohormones not only helps to reduce production costs but also improves the efficiency of biofuel utilization. However, accurate and sensitive phytohormones determination and analytical methods are the basis for plant hormone research. In this study, the characteristics of phytohormones in microalgae and research progress for regulating the accumulation of energy storage substances in microalgae by exogenous phytohormones, combined with abiotic stress conditions at home and abroad, are summarized. The possible metabolic mechanism of phytohormones in microalgae is discussed, and possible future research directions are put forward, which provide a theoretical basis for the application of phytohormones in microalgae. [ABSTRACT FROM AUTHOR]

Polyamines are a series of short-chain aliphatic amines which can be produced during the metabolism of cells, and are widely distributed in aquatic organisms. Polyamines are important regulators for physiological metabolism, which also eliminate active oxygen free radicals, maintain the normal physiological activities of living organisms, and improve plants' resistance to adverse stresses. However, the knowledge of functional characteristics of polyamines in algae, and their effects on the feeding habits of aquatic organisms, are still limited. In this review, the distribution and characteristics of polyamines in unicellular algae are evaluated. Different characteristics of polyamines in various types of algae, and the effects of polyamines being ingested by shellfish, are critically reviewed and analyzed. So far, most literature references offer the opinion that the partial functions of polyamines are similar to plant hormones. Some microalgae diets have different effects on bivalve mollusks, which could be partly due to characteristic changes of polyamines in algae cells, and their adaptability to external environmental pressure. The goal of this review is to discuss possible reasons why microalgae may be suitable for shellfish feeding. This may provide a new application for microalgae development, as a nutritional feed for shellfish. • Polyamines are important regulators and can improve resistance to adverse stresses. • Polyamines are ubiquitous in algae but the cognition of their functions is limited. • Different effects of algae on shellfish are partly due to the changes of polyamines. [ABSTRACT FROM AUTHOR]

[Display omitted] • Nannochloropsis oceanica was a promising source of violaxanthin cycle carotenoids. • Violaxanthin cycle carotenoids were simultaneously produced. • Ethanol precipitation was an effective purification method. • The purities of obtained violaxanthin cycle carotenoids were all more than 92%. The development of integrated co-production of multiple high-purity carotenoids from microalgal cells holds considerable significance for the valorization of microalgae. In this study, the economical microalga Nannochloropsis oceanica was identified as an accumulator of violaxanthin cycle carotenoids, including violaxanthin, antheraxanthin, and zeaxanthin. Notably, a novel and competent approach for the integrated co-production of violaxanthin cycle carotenoids was explored, encompassing four steps: microalgal cultivation, solvent extraction, octadecylsilyl open-column chromatography, and ethanol precipitation. Under optimal co-production conditions, the purities of the obtained violaxanthin, antheraxanthin, and zeaxanthin all exceeded 92%, with total recovery rates of approximately 51%, 40%, and 60%, respectively. Utilizing nuclear magnetic resonance techniques, the purified violaxanthin, antheraxanthin, and zeaxanthin were identified as all- trans -violaxanthin, all- trans -antheraxanthin, and all- trans -zeaxanthin, respectively. This method held significance for the multiproduct biorefinery of the microalga N. oceanica and carried potential future implications for the violaxanthin cycle carotenoids. [ABSTRACT FROM AUTHOR]

Bivalve lectins perform a crucial function in recognition of foreign particles, such as microalgae and pathogenic bacteria. In this study, a novel C-type lectin form Sinonovacula constricta (ScCL) was characterized. The full-length cDNA of ScCL was 1645 bp, encoding a predicted polypeptide of 273 amino acids with one typical carbohydrate-recognition domain. ScCL has the highest similarity and closest phylogenetic relationship with the C-type lectin from Solen grandis. Real-time PCR analysis showed that ScCL was expressed in all tested tissues, with the highest expression in the foot and the lowest expression in hemocytes. Agglutination activity of ScCL was Ca2+-independent. ScCL showed the strongest agglutination on Chlorella vulgaris , the modest agglutination on Platymonas subcordiformis , Nannochloropsis sp., and Thalassiosira pseudonana , the weakest agglutination on Chaetoceros sp., and no agglutination on Isochrysis zhanjiangensis. Meanwhile, agglutination tests and western blot analysis revealed that the recombinant ScCL protein could agglutinate Staphylococcus aureus and Vibrio harveyi , but could not agglutinate Vibrio anguillarum , Bacillus cereus , or Vibrio parahaemolyticus. Furthermore, ScCL had a high binding activity with LPS and mannose, a low binding activity with LTA, and no binding activity with PGN. The expression of ScCL in the gill of S. constricta fed with C. vulgaris and T. pseudonana was significantly increased at 1 and/or 3 h. After injection with S. aureus , the expression of ScCL in the gill was significantly increased at 3, 6, and 24 h. These results indicated that ScCL was involved in food particle recognition and immunity of S. constricta. • A novel C-type lectin from Sinonovacula constricta (ScCL) was characterized. • ScCL protein could bind to several PAMPs and bacteria. • ScCL protein could agglutinate several microalgae and bacteria in Ca2+-independent. • After feeding Chlorella vulgaris and Thalassiosira pseudonana , or after challenged with staphylococcus aureus , the expression of ScCL was up-regulated. • ScCL may be involved in food recognition and innate immunity. [ABSTRACT FROM AUTHOR]

[Display omitted] • Co-culture of the two algae had the highest nutrient and metal removal efficiency. • The higher tolerance of Chlorella to metals due to the higher GSH and SOD activities. • There was no significant biomagnification of the metals in the algae-shell food chain. This research the bait microalgae Chlorella sp. and Isochrysis zhangjiangensis were fed to Ruditapes philippinarum (clams) using two types of cultures, monoculture and co-culture, simulating the conditions, and presence, of common heavy metals Cu2+, Pb2+ and Cd2+ in cultured waters. The effects of microalgae growth, antioxidant activity, and changes in nutrient TN and TP were investigated. The metal contents of the culture solution, algal cells, and four tissues of Ruditapes philippinarum were analyzed. The results showed that the highest nutrient removal efficiency was achieved under the co-culture of three heavy metals, with the removal rates of TN and TP ranging from 47.26% to 51.36% and 78.28% to 87.72%, respectively. The higher tolerance of Chlorella sp. to heavy metals compared to Isochrysis zhangjiangensis was probably due to the higher glutathione and super oxide dimutase activities exhibited by Chlorella sp. cells, which increased by 28% and 40.64%, respectively. Isochrysis zhanjiangensis , Chlorella sp. and their co-culture group had the highest removal rate of Pb2+, which were 20.15%, 26.56% and 29.19%, respectively, followed by Cd2+ and Cu2+. The enrichment of the four tissues from the Ruditapes philippinarum organism for heavy metals, was in the order of visceral mass > gill > coat membrane > muscle. The biotransfer coefficients of the three metals in the microalgae- Ruditapes philippinarum food chain biomagnification ranged from 0.24 to 0.79, which indicated that there was no significant biomagnification of the metals in the transfer process of the food chain. The results of this study reveal the mechanism of heavy metal transport in the algae-shellfish system, which would provide a theoretical basis for improving the ecological environment, and can improve the quality of cultured shellfish products. [ABSTRACT FROM AUTHOR]

[Display omitted] • Integrated preparation of diadinoxanthin and fucoxanthin from Phaeodactylum tricornutum was developed. • Ethanol precipitation was effective for the purification of diadinoxanthin. • The purity of obtained diadinoxanthin and fucoxanthin was more than 95%. Integrated preparation of high-purity carotenoids from marine microalgae using green and efficient methods still faces enormous challenges. In this study, valorization of the economic Phaeodactylum tricornutum using integrated preparation of diadinoxanthin (Ddx) and fucoxanthin (Fx) was explored containing four steps including algae cultivation, solvent extraction, ODS open-column chromatography, and ethanol precipitation for the first time. Several essential key factors were optimized for simultaneously extracting Ddx and Fx from P. tricornutum. ODS open-column chromatography was used to isolate Ddx and Fx. Purification of Ddx and Fx was accomplished using ethanol precipitation. After optimization, the purity of Ddx and Fx was more than 95%, and the total recovery rates of Ddx and Fx were approximately 55% and 85%, respectively. The purified Ddx and Fx were identified as all- trans -diadinoxanthin and all- trans -fucoxanthin, respectively. The antioxidant capacity of the purified Ddx and Fx was assessed using two tests in vitro: DPPH and ABTS radical assays. [ABSTRACT FROM AUTHOR]

Continuous cropping of Fenghua taro leads to poor quality. In this research, two pollution tolerant algae, Spirulina platensis and Chlorella vulgaris , were selected as conditioners for continuous cropping obstacles of taro production. The composition, abundance, and biodiversity of the bacteria and fungi communities in soil rhizosphere of continuous cropping were compared to non-continuous cropping, and studied in-depth. These results showed that the growth of taro during continuous cropping, when irrigated with Chlorella vulgaris and Spirulina platensis, was significantly better than the control group without algal cell irrigation. Accordingly, the plant height, leaf length, leaf area, moisture, carbohydrate, and protein in continuous cropping were 128 cm, 69 cm, 56 cm, 72 g/100 g, 10.8 g/100 g and 2.4 g/100 g, respectively, when watered with Chlorella vulgaris. Furthermore, compared with continuous cropping, the abundance of plant pathogens in the taro rhizosphere, fertilized with Spirulina platensis, were significantly reduced, while the corresponding abundance of soil beneficial bacteria was significantly increased. Specifically, the abundance of beneficial bacteria Ceratobasidium and Pulveroboletus increased, and the pathogens such as Curvularia , Erysiphe , and Clonostachys decreased, during continuous cropping when irrigated with Spirulina platensis. Live microalgae, after wastewater treatment, could be used as potential conditioners to avoid obstacles during continuous cropping of taro production. [Display omitted] • Chlorella vulgaris and Spirulina platensis grew well in swine wastewater. • Spirulina platensis improved the growth of taro with continuous cropping. • Soil microbial community may be the important factor affecting taro growth. • Algae alleviate taro growth obstacles by optimizing the abundance of microbes. • Live microalgae could be used as potential conditioners to taro production. [ABSTRACT FROM AUTHOR]

The successful care and breeding of intertidal shellfish is determined by the variety and quality of the microalgae used as feed. Practice has found obvious feeding selectivity in different intertidal shellfish for diet microalgae, so screening for microalgal species with high food value is a priority for research related to the culture of hatchery shellfish. In this work, the effects of four different microalgae, Chaetoceros calcitrans , Isochrysis galbana , Nannochloropsis oculata and Diacronema viridis , on the growth of juvenile ark shells ( Tegillarca granosa Linnaeus) was studied. The microalgal diets were applied in three different types of treatments: a mixture of all four microalgae, single species, and binary combinations of C. calcitrans or I. galbana with the other individual species. The best feeding effects were observed with the mixture of all four microalgae, and binary algal diets were second best. When fed with single-species diets, the most favorable results were observed with C. calcitrans followed by I. galbana . Single-species diets of D. viridis or N. oculata were the least beneficial. Fatty acid and sterol compositions of the microalgae and juvenile ark shells were analyzed by gas chromatography–mass spectrometry (GC–MS) after feeding, and 32 fatty acids and 12 sterols were identified. The results of a principal components analysis (PCA) of the fatty acid and sterol compositions in juvenile ark shells, using the SIMCA-P + software package, revealed that these contents were affected by their microalgal diets. In the PCA score scatter plots for the first and third components, the samples fed with the full mixture of microalgae were grouped near the samples fed with binary-species diets of C. calcitrans and I. galbana . The samples fed with binary-species diets of C. calcitrans and N. oculata or D. viridis were located near the samples fed with only C. calcitrans , and the samples fed with binary-species diets of I. galbana and N. oculata or D. viridis were near the samples fed with only I. galbana . The samples fed with only N. oculata or D. viridis were located far from the other samples. These results demonstrate that the lipid compositions of juveniles fed with binary-species diets were similar to those of the shells fed with single microalgae, which had a more favorable feeding effect on the shell. This study highlights the feeding selectivity of the shells, and the results shown in the PCA scatter plot based on the fatty acid and sterol compositions demonstrate that the high-quality microalgae candidates for use in hatchery culture can be selected by comparing samples fed with single and mixed microalgae. Statement of relevance This study demonstrated that the fatty acids and sterols of the shellfish were largely determined by its diet microalgae. Based on the characteristics of the lipid composition of the microalgae candidates, by using GC–MS and principal components analysis (PCA), a precise method can be applied in shellfish hatchery culture when screening the potential microalgal diets of high quality. [ABSTRACT FROM AUTHOR]

Abstract: In the present study, we investigated the major lipid changes of six important diet microalgae (Chlorella sp., Nannochloropsis oculata, Nannochloropsis sp., Isochrysis galbana, Phaeodactylum tricornutum Bohlin and Chaetoceros calcitrans) during the entire growth phase using UPLC-Q-TOF MS system. Our results suggested that EPA/AA-rich lipids were largely detected in both Nannochloropsis sp. and N. oculata as a component of diacylglyceryl-N,N,N-trimethylhomoserine (DGTS). Moreover, docosahexaenoic acids (DHAs) were detected only in I. galbana mainly as a component of monogalactosyldiacylglycerol (MGDG) and sulphoquinovosyldiacylglycerol (SQDG). Besides, the growth phase of microalgae could be a useful variable for the content enrichment of EFA-rich lipids. A higher content EPA/AA-rich DGTS could be obtained at the end of stationary phase for Nannochloropsis sp. and N. oculata, and a higher content of DHA-rich lipids could be obtained at the end of stationary phase for I. galbana. However, EPA/AA-rich lipids were mainly detected in both P. tricornutum and C. calcitrans mainly as a component of TAG and MGDG, respectively. In addition, a higher content of EPA-rich TAG could be obtained at the end of the stationary phase for P. tricornutum. A higher content of EPA-rich MGDG could be obtained at the end of stationary phase for C. calcitrans. Taken together, considering the high EFA requirement for artificial rearing of marine organisms, Nannochloropsis sp., N. oculata, P. tricornutum, C. calcitrans and I. galbana would be highly recommended to be harvested at the end of the stationary phase. The distribution characteristics of EFA-rich lipids related to 6 microalgae are described in detail. [Copyright &y& Elsevier]

Sinonovacula constricta is the first marine mollusc demonstrated to possess all fatty acyl desaturase (Fad) and elongase of very long chain fatty acids (Elovl) activities required for long-chain polyunsaturated fatty acids (LC-PUFA) biosynthesis via the Sprecher pathway, providing a good opportunity to investigate the essential fatty acids (FA) for its larval development. In the present study, we analyzed the Fad and Elovl expressions, FA compositions, and the feed effects of three representative microalgae (Chlorella sp. , Chaetoceros calcitrans and Isochrysis galbana) in S. constricta at early developmental stages. The results showed that S. constricta Fad and Elovl were all highly expressed in zygotes, which might occur from maternal transfer. Compared to transcripts of Elovl4 activity, Elovl2/5 was only expressed relatively high in zygotes, indicating that C ≥ 22 LC-PUFA were more important than C ≤ 20 LC-PUFA for S. constricta larval development. The zygotes accumulated abundant FA from parents, especially eicosapentaenoic acid (EPA), but with small amount of docosahexaenoic acid (DHA). Meanwhile, the trochophore larva fed with single or mixed microalgae with I. galbana (rich in DHA) exhibited significantly higher growth and survival rate compared to those fed with single Chlorella sp. (rich in 18:2n-6 and 18:3n-3) and C. calcitrans (rich in EPA) or both of them. Those findings suggested that the highly expressed Fad and Elovl might be largely responsible for DHA biosynthesis, but this capability was limited to meet the developmental requirement and the dietary DHA was necessary. Taken together, our results compellingly demonstrated that DHA was essential for S. constricta larval development, which is important to guide its breeding. • Fad and Elovl expressions were examined in developing S. constricta. • Fatty acid compositions were analyzed in developing S. constricta. • Trochophore larval growth and survival were evaluated by first-feeding diet. • DHA still appeared to be essential for S. constricta larval development. • Microalgae rich in EPA and DHA were critical for S. constricta breeding. [ABSTRACT FROM AUTHOR]