Transcriptome Analysis in Haematococcus pluvialis: Astaxanthin Induction by Salicylic Acid (SA) and Jasmonic Acid (JA).

PubMed

Gao, Zhengquan; Li, Yan; Wu, Guanxun; Li, Guoqiang; Sun, Haifeng; Deng, Suzhen; Shen, Yicheng; Chen, Guoqiang; Zhang, Ruihao; Meng, Chunxiao; Zhang, Xiaowen

2015-01-01

Haematococcus pluvialis is an astaxanthin-rich microalga that can increase its astaxanthin production by salicylic acid (SA) or jasmonic acid (JA) induction. The genetic transcriptome details of astaxanthin biosynthesis were analyzed by exposing the algal cells to 25 mg/L of SA and JA for 1, 6 and 24 hours, plus to the control (no stress). Based on the RNA-seq analysis, 56,077 unigenes (51.7%) were identified with functions in response to the hormone stress. The top five identified subcategories were cell, cellular process, intracellular, catalytic activity and cytoplasm, which possessed 5600 (~9.99%), 5302 (~9.45%), 5242 (~9.35%), 4407 (~7.86%) and 4195 (~7.48%) unigenes, respectively. Furthermore, 59 unigenes were identified and assigned to 26 putative transcription factors (TFs), including 12 plant-specific TFs. They were likely associated with astaxanthin biosynthesis in Haematococcus upon SA and JA stress. In comparison, the up-regulation of differential expressed genes occurred much earlier, with higher transcript levels in the JA treatment (about 6 h later) than in the SA treatment (beyond 24 h). These results provide valuable information for directing metabolic engineering efforts to improve astaxanthin biosynthesis in H. pluvialis.

Enhanced astaxanthin accumulation in Haematococcus pluvialis using high carbon dioxide concentration and light illumination.

PubMed

Christian, David; Zhang, Jun; Sawdon, Alicia J; Peng, Ching-An

2018-05-01

In this study, an economical two-stage method was proposed for the production of natural astaxanthin from Haematococcus pluvialis without a medium replacement step. In stage 1, H. pluvialis were grown under low light illumination until they reached optimal biomass. In stage 2, cells were switched to astaxanthin induction conditions utilizing the combination of high light illumination and elevated carbon dioxide levels (5 or 15%). The introduction of CO 2 altered the C/N balance creating a nutrient deficiency without a change of media. The resulting astaxanthin yield was 2-3 times that of using either stressor alone. This astaxanthin induction method has many advantages over current methods including no medium replacement and a short induction time of less than four days. Copyright © 2018 Elsevier Ltd. All rights reserved.

Characterization of carotenoid hydroxylase gene promoter in Haematococcus pluvialis.

PubMed

Meng, C X; Wei, W; Su, Z- L; Qin, S

2006-10-01

Astaxanthin, a high-value ketocarotenoid is mainly used in fish aquaculture. It also has potential in human health due to its higher antioxidant capacity than beta-carotene and vitamin E. The unicellular green alga Haematococcus pluvialis is known to accumulate astaxanthin in response to environmental stresses, such as high light intensity and salt stress. Carotenoid hydroxylase plays a key role in astaxanthin biosynthesis in H. pluvialis. In this paper, we report the characterization of a promoter-like region (-378 to -22 bp) of carotenoid hydroxylase gene by cloning, sequence analysis and functional verification of its 919 bp 5'-flanking region in H. pluvialis. The 5'-flanking region was characterized using micro-particle bombardment method and transient expression of LacZ reporter gene. Results of sequence analysis showed that the 5'-flanking region might have putative cis-acting elements, such as ABA (abscisic acid)-responsive element (ABRE), C-repeat/dehydration responsive element (C-repeat/DRE), ethylene-responsive element (ERE), heat-shock element (HSE), wound-responsive element (WUN-motif), gibberellin-responsive element (P-box), MYB-binding site (MBS) etc., except for typical TATA and CCAAT boxes. Results of 5' deletions construct and beta-galactosidase assays revealed that a highest promoter-like region might exist from -378 to -22 bp and some negative regulatory elements might lie in the region from -919 to -378 bp. Results of site-directed mutagenesis of a putative C-repeat/DRE and an ABRE-like motif in the promoter-like region (-378 to -22 bp) indicated that the putative C-repeat/DRE and ABRE-like motif might be important for expression of carotenoid hydroxylase gene.

Optimizing gas transfer to improve growth rate of Haematococcus pluvialis in a raceway pond with chute and oscillating baffles.

PubMed

Yang, Zongbo; Cheng, Jun; Li, Ke; Zhou, Junhu; Cen, Kefa

2016-08-01

Up-down chute and oscillating (UCO) baffles were used to generate vortex and oscillating flow field to improve growth rate of Haematococcus pluvialis in a raceway pond. Effects of gas flow rate, solution velocity, and solution depth on solution mass transfer coefficient and mixing time were evaluated using online pH and dissolved oxygen probes. Mass transfer coefficient increased by 1.3 times and mixing time decreased by 33% when UCO baffles were used in the H. pluvialis solution, resulting in an 18% increase in biomass yield with 2% CO2. The H. pluvialis biomass yield further increased to 1.5g/L, and astaxanthin composition accumulated to 29.7mg/L under relatively higher light intensity and salinity. Copyright © 2016 Elsevier Ltd. All rights reserved.

Astaxanthin-Producing Green Microalga Haematococcus pluvialis: From Single Cell to High Value Commercial Products

PubMed Central

Shah, Md. Mahfuzur R.; Liang, Yuanmei; Cheng, Jay J.; Daroch, Maurycy

2016-01-01

Many species of microalgae have been used as source of nutrient rich food, feed, and health promoting compounds. Among the commercially important microalgae, Haematococcus pluvialis is the richest source of natural astaxanthin which is considered as “super anti-oxidant.” Natural astaxanthin produced by H. pluvialis has significantly greater antioxidant capacity than the synthetic one. Astaxanthin has important applications in the nutraceuticals, cosmetics, food, and aquaculture industries. It is now evident that, astaxanthin can significantly reduce free radicals and oxidative stress and help human body maintain a healthy state. With extraordinary potency and increase in demand, astaxanthin is one of the high-value microalgal products of the future.This comprehensive review summarizes the most important aspects of the biology, biochemical composition, biosynthesis, and astaxanthin accumulation in the cells of H. pluvialis and its wide range of applications for humans and animals. In this paper, important and recent developments ranging from cultivation, harvest and postharvest bio-processing technologies to metabolic control and genetic engineering are reviewed in detail, focusing on biomass and astaxanthin production from this biotechnologically important microalga. Simultaneously, critical bottlenecks and major challenges in commercial scale production; current and prospective global market of H. pluvialis derived astaxanthin are also presented in a critical manner. A new biorefinery concept for H. pluvialis has been also suggested to guide toward economically sustainable approach for microalgae cultivation and processing. This report could serve as a useful guide to present current status of knowledge in the field and highlight key areas for future development of H. pluvialis astaxanthin technology and its large scale commercial implementation. PMID:27200009

Stability and changes in astaxanthin ester composition from Haematococcus pluvialis during storage

NASA Astrophysics Data System (ADS)

Miao, Fengping; Geng, Yahong; Lu, Dayan; Zuo, Jincheng; Li, Yeguang

2013-11-01

In this paper, we investigated the effects of temperature, oxygen, antioxidants, and corn germ oil on the stability of astaxanthin from Haematococcus pluvialis under different storage conditions, and changes in the composition of astaxanthin esters during storage using high performance liquid chromatography and spectrophotometry. Oxygen and high temperatures (22-25°C) significantly reduced the stability of astaxanthin esters. Corn germ oil and antioxidants (ascorbic acid and vitamin E) failed to protect astaxanthin from oxidation, and actually significantly increased the instability of astaxanthin. A change in the relative composition of astaxanthin esters was observed after 96 weeks of long-term storage. During storage, the relative amounts of free astaxanthin and astaxanthin monoesters declined, while the relative amount of astaxanthin diesters increased. Thus, the ratio of astaxanthin diester to monoester increased, and this ratio could be used to indicate if astaxanthin esters have been properly preserved. If the ratio is greater than 0.2, it suggests that the decrease in astaxanthin content could be higher than 20%. Our results show that storing algal powder from H. pluvialis or other natural astaxanthin products under vacuum and in the dark below 4°C is the most economical and applicable storage method for the large-scale production of astaxanthin from H. pluvialis. This storage method can produce an astaxanthin preservation rate of at least 80% after 96 weeks of storage.

Effect of drying, storage temperature and air exposure on astaxanthin stability from Haematococcus pluvialis.

PubMed

Ahmed, Faruq; Li, Yan; Fanning, Kent; Netzel, Michael; Schenk, Peer M

2015-08-01

Astaxanthin is a powerful antioxidant with various health benefits such as prevention of age-related macular degeneration and improvement of the immune system, liver and heart function. To improve the post-harvesting stability of astaxanthin used in food, feed and nutraceutical industries, the biomass of the high astaxanthin producing alga Haematococcus pluvialis was dried by spray- or freeze-drying and under vacuum or air at -20°C to 37°C for 20weeks. Freeze-drying led to 41% higher astaxanthin recovery compared to commonly-used spray-drying. Low storage temperature (-20°C, 4°C) and vacuum-packing also showed higher astaxanthin stability with as little as 12.3±3.1% degradation during 20weeks of storage. Cost-benefit analysis showed that freeze-drying followed by vacuum-packed storage at -20°C can generate AUD$600 higher profit compared to spray-drying from 100kgH. pluvialis powder. Therefore, freeze-drying can be suggested as a mild and more profitable method for ensuring longer shelf life of astaxanthin from H. pluvialis. Copyright © 2015 Elsevier Ltd. All rights reserved.

Organization of astaxanthin within oil bodies of Haematococcus pluvialis studied with polarization-dependent harmonic generation microscopy.

PubMed

Tokarz, Danielle; Cisek, Richard; El-Ansari, Omar; Espie, George S; Fekl, Ulrich; Barzda, Virginijus

2014-01-01

Nonlinear optical microscopy was used to image the localization of astaxanthin accumulation in the green alga, Haematococcus pluvialis. Polarization-in, polarization-out (PIPO) second harmonic generation (SHG) and third harmonic generation (THG) microscopy was applied to study the crystalline organization of astaxanthin molecules in light-stressed H. pluvialis in vivo. Since astaxanthin readily forms H- and J-aggregates in aqueous solutions, PIPO THG studies of astaxanthin aggregates contained in red aplanospores were compared to PIPO THG of in vitro self-assembled H- and J-aggregates of astaxanthin. The PIPO THG data clearly showed an isotropic organization of astaxanthin in red aplanospores of H. pluvialis. This is in contrast to the highly anisotropic organization of astaxanthin in synthetic H- and J-aggregates, which showed to be uniaxial. Since carotenoids in vitro preferentially form H- and J-aggregates, but in vivo form a randomly organized structure, this implies that astaxanthin undergoes a different way of packing in biological organisms, which is either due to the unique physical environment of the alga or is controlled enzymatically.

Organization of Astaxanthin within Oil Bodies of Haematococcus pluvialis Studied with Polarization-Dependent Harmonic Generation Microscopy

PubMed Central

Tokarz, Danielle; Cisek, Richard; El-Ansari, Omar; Espie, George S.; Fekl, Ulrich; Barzda, Virginijus

2014-01-01

Nonlinear optical microscopy was used to image the localization of astaxanthin accumulation in the green alga, Haematococcus pluvialis. Polarization-in, polarization-out (PIPO) second harmonic generation (SHG) and third harmonic generation (THG) microscopy was applied to study the crystalline organization of astaxanthin molecules in light-stressed H. pluvialis in vivo. Since astaxanthin readily forms H- and J-aggregates in aqueous solutions, PIPO THG studies of astaxanthin aggregates contained in red aplanospores were compared to PIPO THG of in vitro self-assembled H- and J-aggregates of astaxanthin. The PIPO THG data clearly showed an isotropic organization of astaxanthin in red aplanospores of H. pluvialis. This is in contrast to the highly anisotropic organization of astaxanthin in synthetic H- and J-aggregates, which showed to be uniaxial. Since carotenoids in vitro preferentially form H- and J-aggregates, but in vivo form a randomly organized structure, this implies that astaxanthin undergoes a different way of packing in biological organisms, which is either due to the unique physical environment of the alga or is controlled enzymatically. PMID:25215522

Analysis and enhancement of astaxanthin accumulation in Haematococcus pluvialis.

PubMed

Orosa, M; Franqueira, D; Cid, A; Abalde, J

2005-02-01

The green microalga Haematococcus pluvialis was cultured with different concentrations of NaNO(3) to determine the effect on cell growth and astaxanthin accumulation. The optimum nitrate concentration to obtain astaxanthin and to avoid the cessation of cell division was 0.15 g/l NaNO(3). The ratio chlorophyll a/total carotenoids proved a good physiological indicator of nitrogen deficiency in the cell. The effect of different carbon sources, malonate and acetate, on astaxanthin accumulation was also studied; up to 13 times more carotenoids per cell were accumulated in cultures with malonate than in cultures without this compound. The pigment analysis was performed by a new low toxicity HPLC method capable of separating chlorophylls a and b, carotenes and xanthophylls in a short-period of time, using low volumes of solvents and with an economical price. With this method even echinenone was separated, which had been unsuccessful by any other method.

Determination of astaxanthin in Haematococcus pluvialis by first-order derivative spectrophotometry.

PubMed

Liu, Xiao Juan; Juan, Liu Xiao; Wu, Ying Hua; Hua, Wu Ying; Zhao, Li Chao; Chao, Zhao Li; Xiao, Su Yao; Yao, Xiao Su; Zhou, Ai Mei; Mei, Zhou Ai; Liu, Xin; Xin, Liu

2011-01-01

A highly selective, convenient, and precise method, first-order derivative spectrophotometry, was applied for the determination of astaxanthin in Haematococcus pluvialis. Ethyl acetate and ethanol (1:1, v/v) were found to be the best extraction solvent tested due to their high efficiency and low toxicity compared with nine other organic solvents. Astaxanthin coexisting with chlorophyll and beta-carotene was analyzed by first-order derivative spectrophotometry in order to optimize the conditions for the determination of astaxanthin. The results show that when detected at 432 nm, the interfering substances could be eliminated. The dynamic linear range was 2.0-8.0 microg/mL, with a correlation coefficient of 0.9916. The detection threshold was 0.41 microg/mL. The RSD for the determination of astaxanthin was in the range of 0.01-0.06%; the results of recovery test were 98.1-108.0%. The statistical analysis between first-order derivative spectrophotometry and HPLC by T-testing did not exceed their critical values, revealing no significant differences between these two methods. It was proved that first-order derivative spectrophotometry is a rapid and convenient method for the determination of astaxanthin in H. pluvialis that can eliminate the negative effect resulting from the coexistence of astaxanthin with chlorophyll and beta-carotene.

Comparative transcriptome analysis of Haematococcus pluvialis on astaxanthin biosynthesis in response to irradiation with red or blue LED wavelength.

PubMed

Lee, Changsu; Ahn, Joon-Woo; Kim, Jin-Baek; Kim, Jee Young; Choi, Yoon-E

2018-06-18

The unicellular green microalga Haematococcus pluvialis has the highest content of the natural antioxidant, astaxanthin. Previously, it was determined that astaxanthin accumulation in H. pluvialis could be induced by blue-wavelength irradiation; however, the molecular mechanism remains unknown. The present study aimed to compare the transcriptome of H. pluvialis, with respect to astaxanthin biosynthesis, under the monochromatic red (660 nm) or blue (450 nm) light-emitting diode (LED) irradiation. Among a total of 165,372 transcripts, we identified 67,703 unigenes, of which 2245 and 171 were identified as differentially expressed genes (DEGs) in response to blue and red irradiation, respectively. Interestingly, expressional changes of blue light receptor cryptochromes were detected in response to blue and/or red LED irradiation in H. pluvialis, which may directly and indirectly regulate astaxanthin biosynthesis. In accordance with this observation, expression of the BKT and CHY genes, which are part of the downstream section of the astaxanthin biosynthetic pathway, was significantly upregulated by blue LED irradiation compared with their expression under control white irradiation. Contrastingly, they were downregulated by red LED irradiation. Our transcriptome study provided molecular insights that highlighted the different of responses of H. pluvialis to red and blue irradiation, especially for astaxanthin biosynthesis.

Ethanol induced astaxanthin accumulation and transcriptional expression of carotenogenic genes in Haematococcus pluvialis.

PubMed

Wen, Zewen; Liu, Zhiyong; Hou, Yuyong; Liu, Chenfeng; Gao, Feng; Zheng, Yubin; Chen, Fangjian

2015-10-01

Haematococcus pluvialis is one of the most promising natural sources of astaxanthin. However, inducing the accumulation process has become one of the primary obstacles in astaxanthin production. In this study, the effect of ethanol on astaxanthin accumulation was investigated. The results demonstrated that astaxanthin accumulation occurred with ethanol addition even under low-light conditions. The astaxanthin productivity could reach 11.26 mg L(-1) d(-1) at 3% (v/v) ethanol, which was 2.03 times of that of the control. The transcriptional expression patterns of eight carotenogenic genes were evaluated using real-time PCR. The results showed that ethanol greatly enhanced transcription of the isopentenyl diphosphate (IPP) isomerase genes (ipi-1 and ipi-2), which were responsible for isomerization reaction of IPP and dimethylallyl diphosphate (DMAPP). This finding suggests that ethanol induced astaxanthin biosynthesis was up-regulated mainly by ipi-1 and ipi-2 at transcriptional level, promoting isoprenoid synthesis and substrate supply to carotenoid formation. Thus ethanol has the potential to be used as an effective reagent to induce astaxanthin accumulation in H. pluvialis. Copyright © 2015 Elsevier Inc. All rights reserved.

Effects of CO2 enrichment on primary photochemistry, growth and astaxanthin accumulation in the chlorophyte Haematococcus pluvialis.

PubMed

Chekanov, K; Schastnaya, E; Solovchenko, A; Lobakova, E

2017-06-01

The atmospheric CO 2 level is limiting for growth of phototrophic organisms such as microalgae, so CO 2 enrichment boosts the growth and photosynthesis of microalgal cultures. Still, excessive CO 2 injection might inhibit photosynthesis of microalgae. We investigated the effect of continuous sparging of the cultures of Haematococcus pluvialis BM 1 (IPPAS H-2018) (Chlorophyceae), the richest natural source of the value-added pigment astaxanthin. H. pluvialis cultures with CO 2 -enriched air-gas mixtures (with CO 2 level from the atmospheric to 20%) on growth and astaxanthin accumulation in the microalga. Special attention was paid to photosynthetic activity and non-photochemical excited chlorophyll states quenching in the microalgal cells, which was monitored via chlorophyll fluorescence analysis. We also report on the capability of CO 2 capture by H. pluvialis derived from direct measurements of its elemental carbon content. The beneficial effect of the moderately high (5%) CO 2 levels on the culture growth and astaxanthin accumulation under stress results in a higher overall astaxanthin productivity. However, increase of the CO 2 level to 10% or 20% was deteriorative for growth, photosynthesis and carbon assimilation. The results support the possibility of combining a traditional two-stage H. pluvialis cultivation with CO 2 bio-capture although a dilution of the flue gas before its injection is required. Copyright © 2017. Published by Elsevier B.V.

Dynamic changes of inorganic nitrogen and astaxanthin accumulation in Haematococcus pluvialis

NASA Astrophysics Data System (ADS)

Liu, Jian-Guo; Yin, Ming-Yan; Zhang, Jing-Pu; Liu, Wei; Meng, Zhao-Cai

2002-12-01

This study on dynamic changes of culture color, astaxanthin and chlorophylls, inorganic N including N-NO{3/-}, N-NO{2/-} and N-NH{4/+} in batch culture of Haematococcus pluvialis exposed to different additive nitrate concentration showed (1) ast/chl ratio was over 0.8 for brown and red algae, but was usually less than 0.5 for green and yellow algae; (2) N-NO{3/-}, in general, was unstable and decreased, except for a small unexpected increase in nitrate enriched treatment groups; (3) measurable amounts of N-NO{2/-}, and N-NH{4/+} were observed respectively with three change modes although no external nitrite and ammonia were added into the culture; (4) a non-linear correlation between ast/chl ratio (or color) changes and the levels of N-NO{3/-}, N-NO{2/-}, N-NH{4/+} in H. pluvialis culture; (5) up and down variation of the ast/chl ratio occurred simultaneously with a perceptible color change from yellow to brown (or red) when N-NO{3/-}, N-NO{2/-} and N-NH{4/+} fluctuated around 30, 5, 5, μmol/L respectively; (6) existence of three dynamic modes of N-NO{3/-}, N-NO{2/-} and N-NH{4/+} changes, obviously associated with initial external nitrate; (7) the key level of total inorganic N concentration regulating the above physiological changes during indoor cultivation was about 50 μmol/L; and (8) 0.5 10 mmol/L of nitrate was theoretically conducive to cell growth in batch culture.

Four Different Methods Comparison for Extraction of Astaxanthin from Green Alga Haematococcus pluvialis

PubMed Central

Dong, Shengzhao; Huang, Yi; Zhang, Rui; Wang, Shihui; Liu, Yun

2014-01-01

Haematococcus pluvialis is one of the potent organisms for production of astaxanthin. Up to now, no efficient method has been achieved due to its thick cell wall hindering solvent extraction of astaxanthin. In this study, four different methods, hydrochloric acid pretreatment followed by acetone extraction (HCl-ACE), hexane/isopropanol (6 : 4, v/v) mixture solvents extraction (HEX-IPA), methanol extraction followed by acetone extraction (MET-ACE, 2-step extraction), and soy-oil extraction, were intensively evaluated for extraction of astaxanthin from H. pluvialis. Results showed that HCl-ACE method could obtain the highest oil yield (33.3 ± 1.1%) and astaxanthin content (19.8 ± 1.1%). Quantitative NMR analysis provided the fatty acid chain profiles of total lipid extracts. In all cases, oleyl chains were predominant, and high amounts of polyunsaturated fatty acid chains were observed and the major fatty acid components were oleic acid (13–35%), linoleic acid (37–43%), linolenic acid (20–31%), and total saturated acid (17–28%). DPPH radical scavenging activity of extract obtained by HCl-ACE was 73.2 ± 1.0%, which is the highest amongst the four methods. The reducing power of extract obtained by four extraction methods was also examined. It was concluded that the proposed extraction method of HCl-ACE in this work allowed efficient astaxanthin extractability with high antioxidant properties. PMID:24574909

Four different methods comparison for extraction of astaxanthin from green alga Haematococcus pluvialis.

PubMed

Dong, Shengzhao; Huang, Yi; Zhang, Rui; Wang, Shihui; Liu, Yun

2014-01-01

Haematococcus pluvialis is one of the potent organisms for production of astaxanthin. Up to now, no efficient method has been achieved due to its thick cell wall hindering solvent extraction of astaxanthin. In this study, four different methods, hydrochloric acid pretreatment followed by acetone extraction (HCl-ACE), hexane/isopropanol (6:4, v/v) mixture solvents extraction (HEX-IPA), methanol extraction followed by acetone extraction (MET-ACE, 2-step extraction), and soy-oil extraction, were intensively evaluated for extraction of astaxanthin from H. pluvialis. Results showed that HCl-ACE method could obtain the highest oil yield (33.3±1.1%) and astaxanthin content (19.8±1.1%). Quantitative NMR analysis provided the fatty acid chain profiles of total lipid extracts. In all cases, oleyl chains were predominant, and high amounts of polyunsaturated fatty acid chains were observed and the major fatty acid components were oleic acid (13-35%), linoleic acid (37-43%), linolenic acid (20-31%), and total saturated acid (17-28%). DPPH radical scavenging activity of extract obtained by HCl-ACE was 73.2±1.0%, which is the highest amongst the four methods. The reducing power of extract obtained by four extraction methods was also examined. It was concluded that the proposed extraction method of HCl-ACE in this work allowed efficient astaxanthin extractability with high antioxidant properties.

Biosorption of malachite green onto Haematococcus pluvialis observed through synchrotron-FTIR microspectroscopy.

PubMed

Liu, J H; Zhang, L; Zha, D C; Chen, L Q; Chen, X X; Qi, Z M

2018-06-28

Microalgae have emerged as promising biosorbents for the treatment of malachite green in wastewater. However, the underlying mechanism for the biosorption of malachite green onto microalgae is still unclear and needs further intensive study. In this work, synchrotron Fourier-transform infrared (synchrotron-FTIR) microspectroscoy in combination with biochemical assay is employed to evaluate malachite green removal efficiency (95.2%, 75.6% and 66.5%) by three stages of Haematococcus pluvialis. Meanwhile, the various vital changes of algal cells including lipids, proteins, polysaccharides and carotenoids, is distinguished and quantified in situ. This study illustrates that synchrotron-FTIR microspectroscopy is an effective and powerful tool to scrutinize the mechanism for the interactions between the malachite green dye and microalgal cells and it even provides an effective and none-invasive new approach to screen potentially proper biosorbents for the removal of dyes from wastewater. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.

Isolation and functional identification of a novel cDNA for astaxanthin biosynthesis from Haematococcus pluvialis, and astaxanthin synthesis in Escherichia coli.

PubMed

Kajiwara, S; Kakizono, T; Saito, T; Kondo, K; Ohtani, T; Nishio, N; Nagai, S; Misawa, N

1995-10-01

We succeeded in isolating a novel cDNA involved in astaxanthin biosynthesis from the green alga Haematococcus pluvialis, by an expression cloning method using an Escherichia coli transformant as a host that synthesizes beta-carotene due to the Erwinia uredovora carotenoid biosynthesis genes. The cloned cDNA was shown to encode a novel enzyme, beta-carotene ketolase (beta-carotene oxygenase), which converted beta-carotene to canthaxanthin via echinenone, through chromatographic and spectroscopic analysis of the pigments accumulated in an E. coli transformant. This indicates that the encoded enzyme is responsible for the direct conversion of methylene to keto groups, a mechanism that usually requires two different enzymatic reactions proceeding via a hydroxy intermediate. Northern blot analysis showed that the mRNA was synthesized only in the cyst cells of H. pluvialis. E. coli carrying the H. pluvialis cDNA and the E. uredovora genes required for zeaxanthin biosynthesis was also found to synthesize astaxanthin (3S, 3'S), which was identified after purification by a variety of spectroscopic methods.

The effect of temperature on cell growth and astaxanthin accumulation of Haematococcus pluvialis during a light-dark cyclic cultivation.

PubMed

Wan, Minxi; Zhang, Jingkui; Hou, Dongmei; Fan, Jianhua; Li, Yuanguang; Huang, Jianke; Wang, Jun

2014-09-01

Haematococcus pluvialis is the best source of natural astaxanthin known as "the king of antioxidants". The mass outdoor culture is the most workable strategy for astaxanthin production, but the effects of daytime and night temperatures on the biomass concentration and astaxanthin content of H. pluvialis have received little attention. This study indicated that, raising the daytime or night temperature could stimulate night accumulation of astaxanthin until temperature up to 28°C; the night biomass loss increased firstly and then decreased along with the daytime temperature reducing; decreasing the night temperature can lessen night biomass loss; the daytime temperature of 28°C and the night temperature below 28°C were optimal for achieving high biomass and astaxanthin content. Subsequently, the outdoor culture strategy has been improved and can increase the net biomass and astaxanthin productivities by 5 and 2.9-fold as compared to the former strategy. Copyright © 2014 Elsevier Ltd. All rights reserved.

Variation in Rubisco and other photosynthetic parameters in the life cycle of Haematococcus pluvialis

NASA Astrophysics Data System (ADS)

Chen, Zhangfan; Wang, Guangce; Niu, Jianfeng

2012-01-01

Cells of Haematococcus pluvialis Flot. et Will were collected in four different growth phases. We quantified the initial and total enzyme activity of ribulose-1,5-bisphosphate carboxylase (Rubisco) in crude extracts, and the relative expression of large-subunit ribulose-1,5-bisphosphate caboxylase / oxygenase ( rbcL) mRNA. We measured the ratio of photosynthetic rate to respiration rate (P/R), maximal effective quantum yield of photosystem II ( F v/ F m), electron transport rate (ETR), actual photochemical efficiency of PSII in the light (PSII), and non-photochemical quenching (NPQ). Green vegetative cells were found to be in the most active state, with a relatively higher P/R ratio. These cells also displayed the lowest NPQ and the highest F v/ F m, ETR, and PSII, indicating the most effective PSII. However, both Rubisco activity and rbcL mRNA expression were the lowest measured. In orange resting cysts with relatively lower P/R and NPQ, Rubisco activity and rbcL expression reached a peak, while F v/ F m, ETR, and ΦPSII were the lowest measured. Taking into account the methods of astaxanthin induction used in industry, we suggest that Rubisco may participate in astaxanthin accumulation in H. pluvialis. A continuous and sufficient supply of a carbon source such as CO2 may therefore aid the large scale production of astaxanthin.

Cytotoxic activity of carotenoid rich fractions from Haematococcus pluvialis and Dunaliella salina microalgae and the identification of the phytoconstituents using LC-DAD/ESI-MS.

PubMed

El-Baz, Farouk K; Hussein, Rehab A; Mahmoud, Khaled; Abdo, Sayeda M

2018-02-01

Microalgae represent a rich source that satisfies the growing need for novel ingredients of nutriceuticals, pharmaceuticals, and food supplements. Haematococcus pluvialis and Dunaliella salina microalgae are isolated from the Egyptian hydro-flora and are reported for their potent antioxidant activities. The cytotoxic activity of different fractions of both microalgae was investigated on 4 cell lines HePG2, MCF7, HCT116, and A549. The carotenoid rich fraction of H. pluvialis showed potent cytotoxic activity against colon cancer cell line and moderate activity against both liver and breast cancer cell lines. On the other hand, the carotenoid rich fraction of D. salina showed mild cytotoxic activity on breast and liver cancer cell lines. The carotenoid rich fraction of H. pluvialis was analysed using LC-DAD/ESI-MS and the major carotenoids were identified either free as well as bounded to fatty acids. Copyright © 2017 John Wiley & Sons, Ltd.

Simultaneous measurement of chlorophyll and astaxanthin in Haematococcus pluvialis cells by first-order derivative ultraviolet-visible spectrophotometry.

PubMed

Lababpour, Abdolmajid; Lee, Choul-Gyun

2006-02-01

A first-order derivative spectrophotometric method has been developed for the simultaneous measurement of chlorophyll and astaxanthin concentrations in Haematococcus pluvialis cells. Acetone was selected for the extraction of pigments because of its good sensitivity and low toxicity compared with other organic solvents tested; the tested solvents included acetone, methanol, hexane, chloroform, n-propanol, and acetonitrile. A first-order derivative spectrophotometric method was used to eliminate the effects of the overlaping of the chlorophyll and astaxanthin peaks. The linear ranges in 1D evaluation were from 0.50 to 20.0 microg x ml(-1) for chlorophyll and from 1.00 to 12.0 microg x ml(-1) for astaxanthin. The limits of detection of the analytical procedure were found to be 0.35 microg x ml(-1) for chlorophyll and 0.25 microg x ml(-1) for astaxanthin. The relative standard deviations for the determination of 7.0 microg x ml(-1) chlorophyll and 5.0 microg x ml(-1) astaxanthin were 1.2% and 1.1%, respectively. The procedure was found to be simple, rapid, and reliable. This method was successfully applied to the determination of chlorophyll and astaxanthin concentrations in H. pluvialis cells. A good agreement was achieved between the results obtained by the proposed method and HPLC method.

Low pressure supercritical CO2 extraction of astaxanthin from Haematococcus pluvialis demonstrated on a microfluidic chip.

PubMed

Cheng, Xiang; Qi, ZhenBang; Burdyny, Thomas; Kong, Tian; Sinton, David

2018-02-01

This study demonstrates the efficacy of low pressure supercritical CO 2 extraction of astaxanthin from disrupted Haematococcus pluvialis. A microfluidic reactor was employed that enabled excellent control and allowed direct monitoring of the whole process at the single cell level, in real time. Astaxanthin extraction using ScCO 2 achieved 92% recovery at 55 °C and 8 MPa applied over 15 h. With the addition of co-solvents, ethanol and olive oil, the extraction rates in both experiments were significantly improved reaching full recovery within a few minutes. Notably, for the ethanol case, the timescales of extraction process are reduced 1800-fold from 15 h to 30 s at 55 °C and 8 MPa, representing the fastest complete astaxanthin extraction at such low pressures. Copyright © 2017 Elsevier Ltd. All rights reserved.

Gene expression profiling of astaxanthin and fatty acid pathways in Haematococcus pluvialis in response to different LED lighting conditions.

PubMed

Ma, Ruijuan; Thomas-Hall, Skye R; Chua, Elvis T; Alsenani, Faisal; Eltanahy, Eladl; Netzel, Michael E; Netzel, Gabriele; Lu, Yinghua; Schenk, Peer M

2018-02-01

Haematococcus pluvialis is a green microalga of major interest to industry based on its ability to produce large amounts of astaxanthin. Biosynthesis of astaxanthin and its mono- and di-esters was significantly stimulated under 150 μmol m -2 s -1 of white LED (W-150) compared with lower light intensities, but the highest astaxanthin amounts were produced under 70 μmol m -2 s -1 of blue LED (B-70). Transcripts of astaxanthin biosynthesis genes psy, crtO, and bkt2 were upregulated under W-150, while psy, lcy, crtO, and crtR-B were upregulated by B-70. Total fatty acid content and biosynthesis genes fata and all dgat genes were induced under W-150, while C18:3n6 biosynthesis and dgat2a expression were specifically stimulated by B-70 which was correlated to astaxanthin ester biosynthesis. Nitrogen starvation, various LEDs and the identified upregulated genes may provide useful tools for future metabolic engineering to significantly increase free astaxanthin, its esters and fatty acid precursors in H. pluvialis. Copyright © 2017 Elsevier Ltd. All rights reserved.

Effect of red cyst cell inoculation and iron(II) supplementation on autotrophic astaxanthin production by Haematococcus pluvialis under outdoor summer conditions.

PubMed

Hong, Min-Eui; Choi, Yoon Young; Sim, Sang Jun

2016-01-20

The negative effect of heat stress on the autotrophic astaxanthin production by Haematococcus pluvialis has been observed during outdoor culture in summer. Under the summer conditions, the proliferation of vegetative cells was highly halted in the green stage and the inducibility in the biosynthesis of astaxanthin was partly hindered in the red stage. Herein, under outdoor summer conditions in which variations of the diurnal temperature occur, heat-stress-driven inefficient vegetative growth of H. pluvialis was highly improved by inoculating the red cyst cells; thereby, maintaining relatively moderate intracellular carotenoid levels in the green stage. Subsequently, a remarkably enhanced astaxanthin titer was successfully obtained by supplementing 50 μM iron(II) to induce the heat stress-driven Haber-Weiss reaction in the red stage. As a result, the productivity of astaxanthin in the cells cultured under summer temperature conditions (23.4-33.5 °C) using the two methods of red cell (cyst) inoculation and the iron(Fe(2+)) supplementation was increased by 147% up to 5.53 mg/L day compared with that of the cells cultured under spring temperature conditions (17.5-27.3 °C). Our technical solutions will definitely improve the annual natural astaxanthin productivity in H. pluvialis in locations confronted by hot summer weather, particularly in large-scale closed photobioreactor systems. Copyright © 2015 Elsevier B.V. All rights reserved.

Transcriptome-based analysis on carbon metabolism of Haematococcus pluvialis mutant under 15% CO2.

PubMed

Li, Ke; Cheng, Jun; Lu, Hongxiang; Yang, Weijuan; Zhou, Junhu; Cen, Kefa

2017-06-01

To elucidate the mechanism underlying the enhanced growth rate in the Haematococcus pluvialis mutated with 60 Co-γ rays and domesticated with 15% CO 2 , transcriptome sequencing was conducted to clarify the carbon metabolic pathways of mutant cells. The CO 2 fixation rate of mutant cells increased to 2.57gL -1 d -1 under 15% CO 2 due to the enhanced photosynthesis, carbon fixation, glycolysis pathways. The upregulation of PetH, ATPF0A and PetJ related to photosynthetic electron transport, ATP synthase and NADPH generation promoted the photosynthesis. The upregulation of genes related to Calvin cycle and ppdK promoted carbon fixation in both C3 and C4 photosynthetic pathways. The reallocation of carbon was also enhanced under 15% CO 2 . The 19-, 14- and 3.5-fold upregulation of FBA, TPI and PK genes, respectively, remarkably promoted the glycolysis pathways. This accelerated the conversion of photosynthetic carbon to pyruvate, which was an essential precursor for astaxanthin and lipids biosynthesis. Copyright © 2017 Elsevier Ltd. All rights reserved.

Strain H2-419-4 of Haematococcus pluvialis induced by ethyl methanesulphonate and ultraviolet radiation

NASA Astrophysics Data System (ADS)

Sun, Yanhong; Liu, Jianguo; Zhang, Xiaoli; Lin, Wei

2008-05-01

Two strains H2-410 and H2-419 were obtained from the chemically mutated survivors of wild Haematococcus pluvialis 2 by using ethyl methanesulphonate (EMS). Strains H2-410 and H2-419 showed a fast cell growth with 13% and 20% increase in biomass compared to wild type, respectively. Then H2-419-4, a fast cell growth and high astaxanthin accumulation strain, was obtained by exposing the strain H2-419 to ultraviolet radiation (UV) further. The total biomass, the astaxanthin content per cell, astaxanthin production of H2-419-4 showed 68%, 28%, and 120% increase compared to wild H. pluvialis 2, respectively. HPLC (High Performance Liquid Chromatography) data showed also an obvious proportional variation of different carotenoid compositions in the extracts of H2-419-4 and the wild type, although no peak of carotenoids appeared or disappeared. Therefore, the main compositions in strain H2-419-4, like its wild one, were free of astaxanthin, monoester, and diester of astaxanthin. The asexual reproduction in survivors after exposed to UV was not synchronous, and different from the normal synchronous asexual reproduction as the mother cells were motile instead of non-motile. Interestingly, some survivors from UV irradiation produced many mini-spores (or gamete?), the spores moved away from the mother cell gradually 4 or 5 days later. This is quite similar to sexual reproduction described by Elliot in 1934. However, whether this was sexual reproduction remains questionable, as no mating process has been observed.

Technological process for cell disruption, extraction and encapsulation of astaxanthin from Haematococcus pluvialis.

PubMed

Machado, Francisco R S; Trevisol, Thalles C; Boschetto, Daiane L; Burkert, Janaína F M; Ferreira, Sandra R S; Oliveira, J Vladimir; Burkert, Carlos André V

2016-01-20

In this work, the effectiveness of different enzymatic techniques for cell wall disruption of Haematococcus pluvialis for the extraction of carotenoids and subsequent encapsulation of extracts in the co-polymer poly(hydroxybutyrate-co-hydroxyvalerate) (PHBV) using the Solution Enhanced Dispersion by Supercritical fluids (SEDS) technique was investigated. Glucanex(®) performed best compared with Lyticase(®) and Driselase(®). The conditions for enzymatic lysis using this enzyme preparation were established as a pH of 4.5, a temperature of 55 °C, an initial activity of β-1,3-glucanase of 0.6 U mL(-1) and a reaction time of 30 min. Enzymatic lysis assisted by ultrasound without biomass freezing was shown to be a promising and simple one-step technique for cell wall disruption, reaching 83.90% extractability. In the co-precipitation experiments, the highest encapsulation efficiency (51.21%) was obtained when using a higher biomass to dichloromethane ratio (10 mg mL(-1)) at the carotenoid extraction step and a lower pressure of precipitation (80 bar). In these conditions, spherical particles in the micrometer range (0.228 μm) were obtained. Copyright © 2015 Elsevier B.V. All rights reserved.

Changes of Photosynthetic Behaviors and Photoprotection during Cell Transformation and Astaxanthin Accumulation in Haematococcus pluvialis Grown Outdoors in Tubular Photobioreactors.

PubMed

Zhang, Litao; Su, Fang; Zhang, Chunhui; Gong, Fengying; Liu, Jianguo

2016-12-26

The cell transformation from green motile cells to non-motile cells and astaxanthin accumulation can be induced in the green alga Haematococcus pluvialis cultured outdoors. In the initial 3 d of incubation (cell transformation phase), light absorption and photosynthetic electron transport became more efficient. After five days of incubation (astaxanthin accumulation phase), the light absorption per active reaction center (ABS/RC) increased, but the efficiency of electron transport ( ψ o ) and the quantum yield of electron transport ( φ Eo ) decreased with increased time, indicating that the capacity of photosynthetic energy utilization decreased significantly during astaxanthin accumulation, leading to an imbalance between photosynthetic light absorption and energy utilization. It would inevitably aggravate photoinhibition under high light, e.g., at midday. However, the level of photoinhibition in H. pluvialis decreased as the incubation time increased, which is reflected by the fact that F v / F m determined at midday decreased significantly in the initial 3 d of incubation, but was affected very little after seven days of incubation, compared with that determined at predawn. This might be because the non-photochemical quenching, plastid terminal oxidase, photosystem I cyclic electron transport, defensive enzymes and the accumulated astaxanthin can protect cells against photoinhibition.

Changes of Photosynthetic Behaviors and Photoprotection during Cell Transformation and Astaxanthin Accumulation in Haematococcus pluvialis Grown Outdoors in Tubular Photobioreactors

PubMed Central

Zhang, Litao; Su, Fang; Zhang, Chunhui; Gong, Fengying; Liu, Jianguo

2016-01-01

The cell transformation from green motile cells to non-motile cells and astaxanthin accumulation can be induced in the green alga Haematococcus pluvialis cultured outdoors. In the initial 3 d of incubation (cell transformation phase), light absorption and photosynthetic electron transport became more efficient. After five days of incubation (astaxanthin accumulation phase), the light absorption per active reaction center (ABS/RC) increased, but the efficiency of electron transport (ψo) and the quantum yield of electron transport (φEo) decreased with increased time, indicating that the capacity of photosynthetic energy utilization decreased significantly during astaxanthin accumulation, leading to an imbalance between photosynthetic light absorption and energy utilization. It would inevitably aggravate photoinhibition under high light, e.g., at midday. However, the level of photoinhibition in H. pluvialis decreased as the incubation time increased, which is reflected by the fact that Fv/Fm determined at midday decreased significantly in the initial 3 d of incubation, but was affected very little after seven days of incubation, compared with that determined at predawn. This might be because the non-photochemical quenching, plastid terminal oxidase, photosystem I cyclic electron transport, defensive enzymes and the accumulated astaxanthin can protect cells against photoinhibition. PMID:28035956

Comparative analysis of astaxanthin and its esters in the mutant E1 of Haematococcus pluvialis and other green algae by HPLC with a C30 column.

PubMed

Peng, Juan; Xiang, WenZhou; Tang, QuanMing; Sun, Ni; Chen, Feng; Yuan, JianPing

2008-12-01

A gradient reversed-phase high-performance liquid chromatography (HPLC) method using a C30 column was developed for the simultaneous determination of astaxanthin, astaxanthin monoesters and astaxanthin diesters in the green algae Chlorococcum sp., Chlorella zofingiensis, Haematococcus pluvialis and the mutant E1, which was obtained from the mutagenesis of H. pluvialis by exposure to UV-irradiation and ethyl methanesulphonate (EMS) with subsequent screening using nicotine. The results showed that the contents of total astaxanthins including free astaxanthin and astaxanthin esters ranged from 1.4 to 30.9 mg/g dry biomass in these green algae. The lower total astaxanthin levels (< 2 mg/g dry biomass) were detected in the green algae Chlorococcum sp. and C. zofingiensis. The higher total astaxanthin levels (>16 mg/g dry biomass) were found in the green alga H. pluvialis and its mutant E1. It is notable that the mutant E1 is found to have considerably higher amounts of total astaxanthin (30.9 mg/g) as compared to the wild strain of H. pluvialis (16.1 mg/g). This indicates that UV-irradiation and EMS compound mutagenesis with subsequent screening using nicotine is an effective method for breeding of a high-producing astaxanthin strain of H. pluvialis. In addition, the green alga C. zofingiensis had a remarkably higher percentage of astaxanthin diesters (76.3% of total astaxanthins) and a remarkably lower percentage of astaxanthin monoesters (18.0% of total astaxanthins) in comparison with H. pluvialis (35.5% for diesters and 60.9% for monoesters), the mutant E1 (49.1% and 48.1%) and Chlorococcum sp. (18.0% and 58.6%).

Antioxidative capacity and enzyme activity in Haematococcus pluvialis cells exposed to superoxide free radicals

NASA Astrophysics Data System (ADS)

Liu, Jianguo; Zhang, Xiaoli; Sun, Yanhong; Lin, Wei

2010-01-01

The antioxidative capacity of astaxanthin and enzyme activity of reactive oxygen eliminating enzymes such as superoxide dismutase (SOD), peroxidase (POD), catalase (CAT) and ascorbate peroxidase (APX) were studied in three cell types of Haematococcus pluvialis exposed to high concentrations of a superoxide anion radical (O{2/-}). The results show that defensive enzymes and astaxanthin-related mechanisms were both active in H. pluvialis during exposure to reactive oxygen species (ROS) such as O{2/-}. Astaxanthin reacted with ROS much faster than did the protective enzymes, and had the strongest antioxidative capacity to protect against lipid peroxidation. The defensive mechanisms varied significantly between the three cell types and were related to the level of astaxanthin that had accumulated in those cells. Astaxanthin-enriched red cells had the strongest antioxidative capacity, followed by brown cells, and astaxanthin-deficient green cells. Although there was no significant increase in expression of protective enzymes, the malondialdehyde (MDA) content in red cells was sustained at a low level because of the antioxidative effect of astaxanthin, which quenched O{2/-} before the protective enzymes could act. In green cells, astaxanthin is very low or absent; therefore, scavenging of ROS is inevitably reliant on antioxidative enzymes. Accordingly, in green cells, these enzymes play the leading role in scavenging ROS, and the expression of these enzymes is rapidly increased to reduce excessive ROS. However, because ROS were constantly increased in this study, the enhance enzyme activity in the green cells was not able to repair the ROS damage, leading to elevated MDA content. Of the four defensive enzymes measured in astaxanthin-deficient green cells, SOD eliminates O{2/-}, POD eliminates H2O2, which is a by-product of SOD activity, and APX and CAT are then initiated to scavenge excessive ROS.

Effects of iron electrovalence and species on growth and astaxanthin production of Haematococcus pluvialis

NASA Astrophysics Data System (ADS)

Cai, Minggang; Li, Zhe; Qi, Anxiang

2009-05-01

To increase the cell concentration and the accumulation of astaxanthin in the cultivation of Haematococcus pluvialis, effects of different iron electrovalencies (Fe2+-EDTA and Fe3+-EDTA) and species (Fe-EDTA, Fe(OH){x/32x} and FeC6H5O7) addition on cell growth and accumulation of astaxanthin were studied. Results show that different iron electrovalencies have various effects on cell growth and astaxanthin accumulation of H. pluvialis. Compared with Fe3+-EDTA, Fe2+-EDTA stimulate more effectively the formation of astaxanthin. The maximum astaxanthin content (30.70 mg/g biomass cell) was obtained under conditions of 18 μmol/L Fe2+-EDTA, despite the lower cell density (2.3×105 cell/ml) in such condition. Fe3+-EDTA is more effective than Fe2+-EDTA in improving the cell growth. Especially, the maximal steady-state cell density, 2.9×105 cell/ml was obtained at 18 μmol/L Fe3+-EDTA addition. On the other hand, all the various species of iron (EDTA-Fe, Fe(OH){x/32x}, FeC6H5O7) are capable to improve the growth of the algae and astaxanthin production. Among the three iron species, FeC6H5O7 performed the best. Under the condition of a higher concentration (36 μmol/L) of FeC6H5O7, the cell density and astaxanthin production is 2 and 7 times higher than those of iron-limited group, respectively. The present study demonstrates that the effects of the stimulation with different iron species increased in the order of FeC6H5O7, Fe(OH){x/32x} and EDTA-Fe.

Three-Dimensional Ultrastructural Study of Oil and Astaxanthin Accumulation during Encystment in the Green Alga Haematococcus pluvialis

PubMed Central

Matsuura, Hazuki; Nango, Nobuhito; Hirata, Aiko; Kawano, Shigeyuki

2013-01-01

Haematococcus pluvialis is a freshwater species of green algae and is well known for its accumulation of the strong antioxidant astaxanthin, which is used in aquaculture, various pharmaceuticals, and cosmetics. High levels of astaxanthin are present in cysts, which rapidly accumulate when the environmental conditions become unfavorable for normal cell growth. It is not understood, however, how accumulation of high levels of astaxanthin, which is soluble in oil, becomes possible during encystment. Here, we performed ultrastructural 3D reconstruction based on over 350 serial sections per cell to visualize the dynamics of astaxanthin accumulation and subcellular changes during the encystment of H. pluvialis. This study showcases the marked changes in subcellular elements, such as chloroplast degeneration, in the transition from green coccoid cells to red cyst cells during encystment. In green coccoid cells, chloroplasts accounted for 41.7% of the total cell volume, whereas the relative volume of astaxanthin was very low (0.2%). In contrast, oil droplets containing astaxanthin predominated in cyst cells (52.2%), in which the total chloroplast volume was markedly decreased (9.7%). Volumetric observations also demonstrated that the relative volumes of the cell wall, starch grains, pyrenoids, mitochondria, the Golgi apparatus, and the nucleus in a cyst cell are smaller than those in green coccid cells. Our data indicated that chloroplasts are degraded, resulting in a net-like morphology, but do not completely disappear, even at the red cyst stage. PMID:23326471

Transcriptome Analysis in Haematococcus pluvialis: Astaxanthin Induction by High Light with Acetate and Fe2.

PubMed

He, Bangxiang; Hou, Lulu; Dong, Manman; Shi, Jiawei; Huang, Xiaoyun; Ding, Yating; Cong, Xiaomei; Zhang, Feng; Zhang, Xuecheng; Zang, Xiaonan

2018-01-07

Haematococcus pluvialis is a commercial microalga, that produces abundant levels of astaxanthin under stress conditions. Acetate and Fe 2+ are reported to be important for astaxanthin accumulation in H. pluvialis . In order to study the synergistic effects of high light stress and these two factors, we obtained transcriptomes for four groups: high light irradiation (HL), addition of 25 mM acetate under high light (HA), addition of 20 μM Fe 2+ under high light (HF) and normal green growing cells (HG). Among the total clean reads of the four groups, 156,992 unigenes were found, of which 48.88% were annotated in at least one database (Nr, Nt, Pfam, KOG/COG, SwissProt, KEGG, GO). The statistics for DEGs (differentially expressed genes) showed that there were more than 10 thousand DEGs caused by high light and 1800-1900 DEGs caused by acetate or Fe 2+ . The results of DEG analysis by GO and KEGG enrichments showed that, under the high light condition, the expression of genes related to many pathways had changed, such as the pathway for carotenoid biosynthesis, fatty acid elongation, photosynthesis-antenna proteins, carbon fixation in photosynthetic organisms and so on. Addition of acetate under high light significantly promoted the expression of key genes related to the pathways for carotenoid biosynthesis and fatty acid elongation. Furthermore, acetate could obviously inhibit the expression of genes related to the pathway for photosynthesis-antenna proteins. For addition of Fe 2+ , the genes related to photosynthesis-antenna proteins were promoted significantly and there was no obvious change in the gene expressions related to carotenoid and fatty acid synthesis.

Effects of spray-drying and storage on astaxanthin content of Haematococcus pluvialis biomass.

PubMed

Raposo, Maria Filomena J; Morais, Alcina M M B; Morais, Rui M S C

2012-03-01

The main objective of this study was to evaluate the stability of astaxanthin after drying and storage at different conditions during a 9-week period. Recovery of astaxanthin was evaluated by extracting pigments from the dried powders and analysing extracts by HPLC. The powders obtained were stored under different conditions of temperature and oxygen level and the effects on the degradation of astaxanthin were examined. Under the experimental conditions conducted in this study, the drying temperature that yielded the highest content of astaxanthin was 220°C, as the inlet, and 120°C, as the outlet temperature of the drying chamber. The best results were obtained for biomass dried at 180/110°C and stored at -21°C under nitrogen, with astaxanthin degradation lower than 10% after 9 weeks of storage. A reasonable preservation of astaxanthin can be achieved by conditions 180/80°C, -21°C nitrogen, 180/110°C, 21°C nitrogen, and 220/80°C, 21°C vacuum: the ratio of astaxanthin degradation is equal or inferior to 40%. In order to prevent astaxanthin degradation of Haematococcus pluvialis biomass, it is recommended the storage of the spray dried carotenized cells (180/110ºC) under nitrogen and -21°C.

Astaxanthin from Haematococcus pluvialis Prevents Oxidative Stress on Human Endothelial Cells without Toxicity.

PubMed

Régnier, Philippe; Bastias, Jorge; Rodriguez-Ruiz, Violeta; Caballero-Casero, Noelia; Caballo, Carmen; Sicilia, Dolores; Fuentes, Axelle; Maire, Murielle; Crepin, Michel; Letourneur, Didier; Gueguen, Virginie; Rubio, Soledad; Pavon-Djavid, Graciela

2015-05-07

Astaxanthin, a powerful antioxidant, is a good candidate for the prevention of intracellular oxidative stress. The aim of the study was to compare the antioxidant activity of astaxanthin present in two natural extracts from Haematococcus pluvialis, a microalgae strain, with that of synthetic astaxanthin. Natural extracts were obtained either by solvent or supercritical extraction methods. UV, HPLC-DAD and (HPLC-(atmospheric pressure chemical ionization (APCI)+)/ion trap-MS) characterizations of both natural extracts showed similar compositions of carotenoids, but different percentages in free astaxanthin and its ester derivatives. The Trolox equivalent antioxidant capacity (TEAC) assay showed that natural extracts containing esters displayed stronger antioxidant activities than free astaxanthin. Their antioxidant capacities to inhibit intracellular oxidative stress were then evaluated on HUVEC cells. The intracellular antioxidant activity in natural extracts was approximately 90-times higher than synthetic astaxanthin (5 µM). No modification, neither in the morphology nor in the viability, of vascular human cells was observed by in vitro biocompatibility study up to 10 µM astaxanthin concentrations. Therefore, these results revealed the therapeutic potential of the natural extracts in vascular human cell protection against oxidative stress without toxicity, which could be exploited in prevention and/or treatment of cardiovascular diseases.

Astaxanthin from Haematococcus pluvialis Prevents Oxidative Stress on Human Endothelial Cells without Toxicity

PubMed Central

Régnier, Philippe; Bastias, Jorge; Rodriguez-Ruiz, Violeta; Caballero-Casero, Noelia; Caballo, Carmen; Sicilia, Dolores; Fuentes, Axelle; Maire, Murielle; Crepin, Michel; Letourneur, Didier; Gueguen, Virginie; Rubio, Soledad; Pavon-Djavid, Graciela

2015-01-01

Astaxanthin, a powerful antioxidant, is a good candidate for the prevention of intracellular oxidative stress. The aim of the study was to compare the antioxidant activity of astaxanthin present in two natural extracts from Haematococcus pluvialis, a microalgae strain, with that of synthetic astaxanthin. Natural extracts were obtained either by solvent or supercritical extraction methods. UV, HPLC-DAD and (HPLC-(atmospheric pressure chemical ionization (APCI)+)/ion trap-MS) characterizations of both natural extracts showed similar compositions of carotenoids, but different percentages in free astaxanthin and its ester derivatives. The Trolox equivalent antioxidant capacity (TEAC) assay showed that natural extracts containing esters displayed stronger antioxidant activities than free astaxanthin. Their antioxidant capacities to inhibit intracellular oxidative stress were then evaluated on HUVEC cells. The intracellular antioxidant activity in natural extracts was approximately 90-times higher than synthetic astaxanthin (5 µM). No modification, neither in the morphology nor in the viability, of vascular human cells was observed by in vitro biocompatibility study up to 10 µM astaxanthin concentrations. Therefore, these results revealed the therapeutic potential of the natural extracts in vascular human cell protection against oxidative stress without toxicity, which could be exploited in prevention and/or treatment of cardiovascular diseases. PMID:25962124

Transcriptome sequencing and metabolic pathways of astaxanthin accumulated in Haematococcus pluvialis mutant under 15% CO2.

PubMed

Cheng, Jun; Li, Ke; Zhu, Yanxia; Yang, Weijuan; Zhou, Junhu; Cen, Kefa

2017-03-01

Transcriptome sequencing and annotation was performed on Haematococcus pluvialis mutant red cells induced with high light under 15% CO 2 to demonstrate why astaxanthin yield of the mutant was 1.7 times higher than that of a wild strain. It was found that 56% of 1947 differentially expressed genes were upregulated in mutant cells. Most significant differences were found in unigenes related to photosynthesis, carotenoid biosynthesis and fatty acid biosynthesis pathways. The pyruvate kinase increased by 3.5-fold in mutant cells. Thus, more pyruvate, which was beneficial to carotenoids and fatty acid biosynthesis, was generated. Phytoene synthase, zeta-carotene desaturase, lycopene beta-cyclase involved in β-carotene biosynthesis in mutant cells were upregulated by 10.4-, 4.4-, and 5.8-fold, respectively. Beta-carotene 3-hydroxylase catalyzing conversion of β-carotene into astaxanthin was upregulated by 18.4-fold. The fatty acid biosynthesis was promoted because of the upregulation of acetyl-CoA synthetase and acetyl-CoA carboxylase, thus increasing astaxanthin esterification and accumulation in mutant cells. Copyright © 2016 Elsevier Ltd. All rights reserved.

Simultaneous determination of 13 carotenoids by a simple C18 column-based ultra-high-pressure liquid chromatography method for carotenoid profiling in the astaxanthin-accumulating Haematococcus pluvialis.

PubMed

Jin, Hui; Lao, Yong Min; Zhou, Jin; Zhang, Huai Jin; Cai, Zhong Hua

2017-03-10

A simple ultra-high-pressure liquid chromatography (UHPLC) method for rapidly and simultaneously identifying thirteen carotenoids in Haematococcus pluvialis was developed in this study. The method is capable of effectively separating two astaxanthin isomers, two ζ-carotene isomers, and three phytoene isomers on two simple C18 columns within 9 and 12min only by using methanol and acetonitrile, respectively. To our best knowledge, this is the rapidest method for these carotenoid isomers, currently. Using this method, carotenoid profiling in the astaxanthin-accumulating H. pluvialis under environmental stresses was successfully carried out. Results indicated that carotenoid biosynthesis was differentially perturbed by environmental stresses, indicating that this simple and rapid method is suitable to not only bacterial but also algal samples, with potential applications for a wide range of samples from plant to animal. Finally, possible reasons for the elution order of carotenoids were studied. Copyright © 2017 Elsevier B.V. All rights reserved.

Hydrolysis kinetics of astaxanthin esters and stability of astaxanthin of Haematococcus pluvialis during saponification.

PubMed

Yuan, J P; Chen, F

1999-01-01

The reaction kinetics for the hydrolysis of astaxanthin esters and the degradation of astaxanthin during saponification of the pigment extract from the microalga Haematococcus pluvialis were investigated. Different concentrations of sodium hydroxide in methanol were used for the saponification under nitrogen in darkness at ambient temperature (22 degrees C) followed by the analysis of astaxanthins and other carotenoids using an HPLC method. The concentration of methanolic NaOH solution was important for promoting the hydrolysis of astaxanthin esters and minimizing the degradation of astaxanthin during saponification. With a higher concentration of methanolic NaOH solution, the reaction rate of hydrolysis was high, but the degradation of astaxanthin occurred significantly. The rate constants of the hydrolysis reaction (first order) of astaxanthin esters and the degradation reaction (zero-order) of astaxanthin were directly proportional to the concentration of sodium hydroxide in the saponified solution. Although the concentration of sodium hydroxide in the saponified solution was 0.018 M, complete hydrolysis of astaxanthin esters was achieved in 6 h for different concentrations (10-100 mg/L) of pigment extracts. Results also indicated that a higher temperature should be avoided to minimize the degradation of astaxanthin. In addition, during saponification, no loss of lutein, beta-carotene, and canthaxanthin was found.

Photo Protection of Haematococcus pluvialis Algae by Astaxanthin: Unique Properties of Astaxanthin Deduced by EPR, Optical and Electrochemical Studies

PubMed Central

Focsan, A. Ligia; Polyakov, Nikolay E.; Kispert, Lowell D.

2017-01-01

The antioxidant astaxanthin is known to accumulate in Haematococcus pluvialis algae under unfavorable environmental conditions for normal cell growth. The accumulated astaxanthin functions as a protective agent against oxidative stress damage, and tolerance to excessive reactive oxygen species (ROS) is greater in astaxanthin-rich cells. The detailed mechanisms of protection have remained elusive, however, our Electron Paramagnetic Resonance (EPR), optical and electrochemical studies on carotenoids suggest that astaxanthin’s efficiency as a protective agent could be related to its ability to form chelate complexes with metals and to be esterified, its inability to aggregate in the ester form, its high oxidation potential and the ability to form proton loss neutral radicals under high illumination in the presence of metal ions. The neutral radical species formed by deprotonation of the radical cations can be very effective quenchers of the excited states of chlorophyll under high irradiation. PMID:29065482

In vivo kinetics of lipids and astaxanthin evolution in Haematococcus pluvialis mutant under 15% CO2 using Raman microspectroscopy.

PubMed

Li, Ke; Cheng, Jun; Ye, Qing; He, Yong; Zhou, Junhu; Cen, Kefa

2017-11-01

In vivo spatiotemporal dynamics of lipids and astaxanthin evolution in Haematococcus pluvialis mutant induced with 15% CO 2 and high light intensity were monitored with high spatial resolution in a non-destructive and label-free manner using single-cell Raman imaging. Astaxanthin intensity increased by 3.5 times within 12h under 15% CO 2 , and the accumulation rate was 5.8 times higher than that under air. Lipids intensity under 15% CO 2 was 27% higher than that under air. The lipids initially concentrated in chloroplast under 15% CO 2 due to an increase of directly photosynthetic fatty acid, which was different from the whole-cell dispersed lipids under air. Astaxanthin produced in chloroplast first accumulated around nucleus and then spread in cytoplasmic lipids under both air and 15% CO 2 . The calculation results of kinetic models for lipids and astaxanthin evolutions showed that accumulation rate of lipids was much higher than that of astaxanthin in cells. Copyright © 2017 Elsevier Ltd. All rights reserved.

Media Screening for Obtaining Haematococcus pluvialis Red Motile Macrozooids Rich in Astaxanthin and Fatty Acids

PubMed Central

Butler, Thomas O.; McDougall, Gordon J.; Campbell, Raymond; Stanley, Michele S.; Day, John G.

2017-01-01

Astaxanthin from Haematococcus pluvialis is commercially produced in a two-stage process, involving green vegetative (macrozooid) and red aplanospore stages. This approach has been scaled up to an industrial process but constraints limit its commercial success and profitability, including: contamination issues, high pigment extraction costs, requirements for high light levels and photo-bleaching in the red stage. However, in addition to the aplanospore stage, this alga can produce astaxanthin in vegetative palmelloid and motile macrozooid cells. In this study, a two-stage process utilising different media in the green stage, with subsequent re-suspension in medium without nitrate was employed to optimise the formation of red motile macrozooids. Optimal growth in the green phase was obtained on cultivation under mixotrophic conditions in EG:JM media followed by re-suspension in medium without nitrate resulting in red motile macrozooids with an astaxanthin content of 2.74% (78.4% of total carotenoids) and a lipid content of 35.3% (rich in unsaturated fatty acids. It is envisaged that the red motile macrozooids could be harvested and fed as a whole-cell product directly in the animal feed and aquaculture sectors, or used as a blend of carotenoids and polyunsaturated fatty acids (PUFAs) in nutraceutical products. PMID:29278377

Media Screening for Obtaining Haematococcus pluvialis Red Motile Macrozooids Rich in Astaxanthin and Fatty Acids.

PubMed

Butler, Thomas O; McDougall, Gordon J; Campbell, Raymond; Stanley, Michele S; Day, John G

2017-12-26

Astaxanthin from Haematococcus pluvialis is commercially produced in a two-stage process, involving green vegetative (macrozooid) and red aplanospore stages. This approach has been scaled up to an industrial process but constraints limit its commercial success and profitability, including: contamination issues, high pigment extraction costs, requirements for high light levels and photo-bleaching in the red stage. However, in addition to the aplanospore stage, this alga can produce astaxanthin in vegetative palmelloid and motile macrozooid cells. In this study, a two-stage process utilising different media in the green stage, with subsequent re-suspension in medium without nitrate was employed to optimise the formation of red motile macrozooids. Optimal growth in the green phase was obtained on cultivation under mixotrophic conditions in EG:JM media followed by re-suspension in medium without nitrate resulting in red motile macrozooids with an astaxanthin content of 2.74% (78.4% of total carotenoids) and a lipid content of 35.3% (rich in unsaturated fatty acids. It is envisaged that the red motile macrozooids could be harvested and fed as a whole-cell product directly in the animal feed and aquaculture sectors, or used as a blend of carotenoids and polyunsaturated fatty acids (PUFAs) in nutraceutical products.

Modulation of photosynthetic activity and photoprotection in Haematococcus pluvialis cells during their conversion into haematocysts and back.

PubMed

Chekanov, Konstantin; Lukyanov, Alexander; Boussiba, Sammy; Aflalo, Claude; Solovchenko, Alexei

2016-06-01

The engagement of different photoprotective mechanisms in the cells of the carotenogenic astaxanthin-accumulating chlorophyte Haematococcus pluvialis (i) under favorable conditions, (ii) in the course of stress-induced haematocyst formation and (iii) during recovery from the stress was studied. To this end, we followed the changes in primary photochemistry, electron flow at the acceptor side of photosystem II, and non-photochemical quenching (NPQ) using PAM chlorophyll fluorimetry. A general trend recorded in the stressed cells undergoing transition to haematocysts (and reversed during recovery from the stress) was a gradual reduction of the photosynthetic apparatus accompanied by down-regulation of energy-dependent photoprotective mechanisms such as NPQ, along with the accumulation of astaxanthin. On this background, a transient up-regulation of the photosynthetic activity was detected at the intermediated stages (20-50 h of the stress exposure) of haematocyst formation. This phenomenon was tentatively related with the peak of metabolic activity found earlier in the forming haematocysts. The role of secondary carotenogenesis coupled with a reversible transition from 'active' (energy-dependent) to 'passive' photoprotective mechanisms in the extremely high stress tolerance of carotenogenic phototrophs is discussed.

Identification of optimum fatty acid extraction methods for two different microalgae Phaeodactylum tricornutum and Haematococcus pluvialis for food and biodiesel applications.

PubMed

Otero, Paz; Saha, Sushanta Kumar; Gushin, Joanne Mc; Moane, Siobhan; Barron, John; Murray, Patrick

2017-07-01

Microalgae have the potential to synthesize and accumulate lipids which contain high value fatty acids intended for nutrition and biodiesel applications. Nevertheless, lipid extraction methods for microalgae cells are not well established and there is not a standard analytical methodology to extract fatty acids from lipid-producing microalgae. In this paper, current lipid extraction procedures employing organic solvents (chloroform/methanol, 2:1 and 1:2, v/v), sodium hypochlorite solution (NaClO), acid-catalysed hot-water extraction and the saponification process [2.5 M KOH/methanol (1:4, v/v)] have been evaluated with two species of microalgae with different types of cell walls. One is a marine diatom, Phaeodactylum tricornutum, and the other a freshwater green microalga, Haematococcus pluvialis. Lipids from all types of extracts were estimated gravimetrically and their fatty acids were quantified by a HPLC equipped with Q-TOF mass spectrometer. Results indicated significant differences both in lipids yield and fatty acids composition. The chloroform and methanol mixture was the most effective extraction solvent for the unsaturated fatty acids such as DPA (C22:05), DHA, (C22:06), EPA (C20:05) and ARA (C20:04). While acid treatments improved the saturated fatty acids (SFAs) yield, especially the short chain SFA, lauric acid (C12:0), whose amount was 64% higher in P. tricornutum and 156% higher in H. pluvialis compared to organic solvent extractions. Graphical abstract ᅟ.

A new paradigm for producing astaxanthin from the unicellular green alga Haematococcus pluvialis.

PubMed

Zhang, Zhen; Wang, Baobei; Hu, Qiang; Sommerfeld, Milton; Li, Yuanguang; Han, Danxiang

2016-10-01

The unicellular green alga Haematococcus pluvialis has been exploited as a cell factory to produce the high-value antioxidant astaxanthin for over two decades, due to its superior ability to synthesize astaxanthin under adverse culture conditions. However, slow vegetative growth under favorable culture conditions and cell deterioration or death under stress conditions (e.g., high light, nitrogen starvation) has limited the astaxanthin production. In this study, a new paradigm that integrated heterotrophic cultivation, acclimation of heterotrophically grown cells to specific light/nutrient regimes, followed by induction of astaxanthin accumulation under photoautotrophic conditions was developed. First, the environmental conditions such as pH, carbon source, nitrogen regime, and light intensity, were optimized to induce astaxanthin accumulation in the dark-grown cells. Although moderate astaxanthin content (e.g., 1% of dry weight) and astaxanthin productivity (2.5 mg L(-1)  day(-1) ) were obtained under the optimized conditions, a considerable number of cells died off when subjected to stress for astaxanthin induction. To minimize the susceptibility of dark-grown cells to light stress, the algal cells were acclimated, prior to light induction of astaxanthin biosynthesis, under moderate illumination in the presence of nitrogen. Introduction of this strategy significantly reduced the cell mortality rate under high-light and resulted in increased cellular astaxanthin content and astaxanthin productivity. The productivity of astaxanthin was further improved to 10.5 mg L(-1)  day(-1) by implementation of such a strategy in a bubbling column photobioreactor. Biochemical and physiological analyses suggested that rebuilding of photosynthetic apparatus including D1 protein and PsbO, and recovery of PSII activities, are essential for acclimation of dark-grown cells under photo-induction conditions. Biotechnol. Bioeng. 2016;113: 2088-2099. © 2016 The Authors

A new paradigm for producing astaxanthin from the unicellular green alga Haematococcus pluvialis

PubMed Central

Zhang, Zhen; Wang, Baobei; Hu, Qiang; Sommerfeld, Milton

2016-01-01

ABSTRACT The unicellular green alga Haematococcus pluvialis has been exploited as a cell factory to produce the high‐value antioxidant astaxanthin for over two decades, due to its superior ability to synthesize astaxanthin under adverse culture conditions. However, slow vegetative growth under favorable culture conditions and cell deterioration or death under stress conditions (e.g., high light, nitrogen starvation) has limited the astaxanthin production. In this study, a new paradigm that integrated heterotrophic cultivation, acclimation of heterotrophically grown cells to specific light/nutrient regimes, followed by induction of astaxanthin accumulation under photoautotrophic conditions was developed. First, the environmental conditions such as pH, carbon source, nitrogen regime, and light intensity, were optimized to induce astaxanthin accumulation in the dark‐grown cells. Although moderate astaxanthin content (e.g., 1% of dry weight) and astaxanthin productivity (2.5 mg L−1 day−1) were obtained under the optimized conditions, a considerable number of cells died off when subjected to stress for astaxanthin induction. To minimize the susceptibility of dark‐grown cells to light stress, the algal cells were acclimated, prior to light induction of astaxanthin biosynthesis, under moderate illumination in the presence of nitrogen. Introduction of this strategy significantly reduced the cell mortality rate under high‐light and resulted in increased cellular astaxanthin content and astaxanthin productivity. The productivity of astaxanthin was further improved to 10.5 mg L−1 day−1 by implementation of such a strategy in a bubbling column photobioreactor. Biochemical and physiological analyses suggested that rebuilding of photosynthetic apparatus including D1 protein and PsbO, and recovery of PSII activities, are essential for acclimation of dark‐grown cells under photo‐induction conditions. Biotechnol. Bioeng. 2016;113: 2088–2099. �

Sodiation as a tool for enhancing the diagnostic value of MALDI-TOF/TOF-MS spectra of complex astaxanthin ester mixtures from Haematococcus pluvialis.

PubMed

Weesepoel, Yannick; Vincken, Jean-Paul; Pop, Raluca Maria; Liu, Kun; Gruppen, Harry

2013-07-01

The microalga Haematococcus pluvialis produces the pigment astaxanthin mainly in esterified form with a multitude of fatty acids, which results in a complex mixture of carotenol mono- and diesters. For rapid fingerprinting of these esters, matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF/TOF-MS) might be an alternative to traditional chromatographic separation combined with MS. Investigation of ionization and fragmentation of astaxanthin mono- and diester palmitate standards in MALDI-TOF/TOF-MS showed that sodium adduct parent masses [M + Na](+) gave much simpler MS(2) spectra than radical / protonated [M](+●) / [M + H](+) parents. [M + Na](+) fragments yielded diagnostic polyene-specific eliminations and fatty acid neutral losses, whereas [M](+●) / [M + H](+) fragmentation resulted in a multitude of non-diagnostic daughters. For diesters, a benzonium fragment, formed by polyene elimination, was required for identification of the second fatty acid attached to the astaxanthin backbone. Parents were forced into [M + Na](+) ionization by addition of sodium acetate, and best signal-to-noise ratios were obtained in the 0.1 to 1.0 mM range. This method was applied to fingerprinting astaxanthin esters in a crude H. pluvialis extract. Prior to MALDI-TOF/TOF-MS, the extract was fractionated by normal phase Flash chromatography to obtain fractions enriched in mono- and diesters and to remove pheophytin a, which compromised monoester signals. All 12 types of all-trans esterified esters found in LC were identified with MALDI-TOF/TOF-MS, with the exception of two minor monoesters. Copyright © 2013 John Wiley & Sons, Ltd.

Effects of temperature on the astaxanthin productivity and light harvesting characteristics of the green alga Haematococcus pluvialis.

PubMed

Giannelli, Luca; Yamada, Hiroyuki; Katsuda, Tomohisa; Yamaji, Hideki

2015-03-01

The green alga Haematococcus pluvialis, which accumulates astaxanthin at an optimal temperature of 20°C, was cultivated under temperatures of 20°C, 23.5°C, 27°C, and 30.5°C, in order to assess the effects on algal metabolism during the growth phase. The culture growth rate declined with above-optimal increases in temperature, and the final maximum cell concentration at 30.5°C reached only 35% of that attained at 20°C. On the contrary, the biomass productivity was increased under all the high-temperature conditions, probably reflecting the metabolism switch from cell duplication to energy accumulation that is typically observed in algal cultures subjected to environmental stress. Moreover, an increase in the light-harvesting capability of the alga was observed by means of the total pigment balance and the photosynthesis-intensity (PI) curve measured under the different cultivation conditions. Cultures kept at higher temperatures were able to better harvest and utilize the impinging light due to photo-acclimation. Finally, the differences in the astaxanthin metabolism were elucidated by subjecting the cultures to nitrogen starvation at 20°C and 27°C. In the culture at 27°C, a 1.4-fold increase in the astaxanthin productivity was observed when compared to that at 20°C, and the latter required almost two-fold more energy for the astaxanthin production compared with the 27°C culture. Copyright © 2014. Published by Elsevier B.V.

Determination of astaxanthin and astaxanthin esters in the microalgae Haematococcus pluvialis by LC-(APCI)MS and characterization of predominant carotenoid isomers by NMR spectroscopy.

PubMed

Holtin, Karsten; Kuehnle, Maximilian; Rehbein, Jens; Schuler, Paul; Nicholson, Graeme; Albert, Klaus

2009-11-01

The oily product ZANTHIN consists of natural astaxanthin, which is manufactured from the microalgae Haematococcus pluvialis by supercritical CO(2) extraction. An HPLC method was developed to separate all of the components of the complex astaxanthin extract using a C(30) column. The separation resulted in different isomers of astaxanthin accompanied by two other carotenoids. The main component consisted of astaxanthin singly esterified with several different fatty acids. C18:3, C18:2, C18:1 and C16:0 were identified as the most commonly occurring fatty acids. Doubly esterified astaxanthin was also found, although in lower concentrations compared to singly esterified astaxanthin. After performing a detailed fatty acid analysis by GC-MS, the peaks from the extract were assigned via HPLC-MS. A trans to cis transmutation of the all-trans compound was performed by thermal treatment in order to obtain an enrichment of cis isomers as the basis for unambiguous identification via NMR experiments. The all-trans as well as the 9- and 13-cis isomers of astaxanthin were characterized in detail by UV/Vis, (1)H, and (1)H,(1)H COSY NMR spectroscopy.

Identification and quantification of astaxanthin esters in shrimp (Pandalus borealis) and in a microalga (Haematococcus pluvialis) by liquid chromatography-mass spectrometry using negative ion atmospheric pressure chemical ionization.

PubMed

Breithaupt, Dietmar E

2004-06-16

Negative ion liquid chromatography-atmospheric pressure chemical ionization mass spectrometry [negative ion LC-(APCI)MS] was used for the identification of astaxanthin esters in extracts of commercial shrimp (Pandalus borealis) and dried microalga (Haematococcus pluvialis) samples. A cleanup step using a normal phase solid phase extraction (SPE) cartridge was applied prior to analysis. Recovery experiments with astaxanthin oleate as model compound proved the applicability of this step (98.5 +/- 7.6%; n = 4). The assignment of astaxanthin esters in negative ion LC-(APCI)MS was based on the detection of the molecular ion (M*-) and the formation of characteristic fragment ions, resulting from the loss of one or two fatty acids. Quantification of individual astaxanthin esters was performed using an astaxanthin calibration curve, which was found to be linear over the required range (1-51 micromol/L; r2 = 0.9996). Detection limits, based on the intensity of M*-, a signal-to-noise ratio of 3:1, and an injection volume of 20 microL, were estimated to be 0.05 microg/mL (free astaxanthin), 0.28 microg/mL (astaxanthin-C16:0), and 0.78 microg/mL (astaxanthin-C16:0/C16:0), respectively. This LC-(APCI)MS method allows for the first time the characterization of native astaxanthin esters in P. borealis and H. pluvialis without using time-consuming isolation steps with subsequent gas chromatographic analyses of fatty acid methyl esters. The results suggest that the pattern of astaxanthin-bound polyunsaturated fatty acids of P. borealis does not reflect the respective fatty acid pattern found in triacylglycerides. Application of the presented LC-(APCI)MS technique in common astaxanthin ester analysis will forestall erroneous xanthophyll ester assignment in natural sources.

Some Physical Parameters to Effect the Production of Heamatococcus pluvialis

NASA Astrophysics Data System (ADS)

Akpolat, O.; Eristurk, S.

The aim of this study is to optimize the physical parameters affecting the production of Haematococcus pluvialis in photobioreactors and to simulate the process. Heamatococcus pluvialis is a green microalgea to have a great interest for production of natural astaxanthin and it can be cultivated in a closed photobiorector system under controlled conditions. Biomass composition, growth rate and high value product spectra like polyunsaturated fatty acids, pigments, poly saccariydes or vitamins depend on strongly the parameters of cultivation process. These are composition of cultivation medium, mixing model and aeration rate, hydrodynamic stress of medium which can be changed by adding some chemicals, cultivation temperature, pH, carbon dioxide and oxygen supply and most important of all: illumination. One of the most important problems during the cultivation is that cells have sensitivity to shear stress very much and the shear stress created by aeration and mixing effects the growth rate of the cell negatively and decreases yield. In this study, physical parameters such as; the rate of the air fed into the reactor, the mixing type, the reduction of the hydrodynamic stress by CMC addition, the effect of the cell size on the cell production and the flocculation speed of the culture, were investigated.

A Combination of Soybean and Haematococcus Extract Alleviates Ultraviolet B-Induced Photoaging

PubMed Central

Shin, Jieun; Kim, Jong-Eun; Pak, Kum-Ju; Kang, Jung Il; Kim, Tae-Seok; Lee, Sang-Yoon; Yeo, Ik-Hyun; Park, Jung Han Yoon; Kim, Jong Hun; Kang, Nam Joo; Lee, Ki Won

2017-01-01

Soybean-derived isoflavones have been investigated for their preventative effects against UV-induced symptoms of skin damage including wrinkle formation and inflammation. Haematococcus pluvialis is a freshwater species of Chlorophyta that contains high concentrations of the natural carotenoid pigment astaxanthin. Astaxanthin is known to be involved in retinoic acid receptor (RAR) signaling and previously been associated with the inhibition of activator protein (AP)-1 dependent transcription. Based on previous studies, we hypothesized that a combination of soy extract (SE) and Haematococcus extract (HE) may prevent UVB-induced photoaging through specific signaling pathways, as measured by UVB-induced wrinkling on hairless mice skin and expression changes in human dermal fibroblasts (HDFs). The 1:2 ratio of SE and HE mixture (SHM) showed the optimal benefit in vivo. SHM was found to inhibit wrinkle formation via the downregulation of matrix metalloproteinase (MMP)-1 mRNA and protein expression. SHM also inhibited mitogen-activated protein kinase (MAPK) phosphorylation and the transactivation of AP-1 which plays an important role in regulating MMP expression. These results highlight the potential for SHM to be developed as a therapeutic agent to prevent UVB-induced skin wrinkling. PMID:28327532

A Combination of Soybean and Haematococcus Extract Alleviates Ultraviolet B-Induced Photoaging.

PubMed

Shin, Jieun; Kim, Jong-Eun; Pak, Kum-Ju; Kang, Jung Il; Kim, Tae-Seok; Lee, Sang-Yoon; Yeo, Ik-Hyun; Park, Jung Han Yoon; Kim, Jong Hun; Kang, Nam Joo; Lee, Ki Won

2017-03-22

Soybean-derived isoflavones have been investigated for their preventative effects against UV-induced symptoms of skin damage including wrinkle formation and inflammation. Haematococcus pluvialis is a freshwater species of Chlorophyta that contains high concentrations of the natural carotenoid pigment astaxanthin. Astaxanthin is known to be involved in retinoic acid receptor (RAR) signaling and previously been associated with the inhibition of activator protein (AP)-1 dependent transcription. Based on previous studies, we hypothesized that a combination of soy extract (SE) and Haematococcus extract (HE) may prevent UVB-induced photoaging through specific signaling pathways, as measured by UVB-induced wrinkling on hairless mice skin and expression changes in human dermal fibroblasts (HDFs). The 1:2 ratio of SE and HE mixture (SHM) showed the optimal benefit in vivo. SHM was found to inhibit wrinkle formation via the downregulation of matrix metalloproteinase (MMP)-1 mRNA and protein expression. SHM also inhibited mitogen-activated protein kinase (MAPK) phosphorylation and the transactivation of AP-1 which plays an important role in regulating MMP expression. These results highlight the potential for SHM to be developed as a therapeutic agent to prevent UVB-induced skin wrinkling.

Enriched Astaxanthin Extract from Haematococcus pluvialis Augments Growth Factor Secretions to Increase Cell Proliferation and Induces MMP1 Degradation to Enhance Collagen Production in Human Dermal Fibroblasts

PubMed Central

Chou, Hsin-Yu; Lee, Chelsea; Pan, Jian-Liang; Wen, Zhi-Hong; Huang, Shu-Hung; Lan, Chi-Wei John; Liu, Wang-Ta; Hour, Tzyh-Chyuan; Hseu, You-Cheng; Hwang, Byeong Hee; Cheng, Kuo-Chen; Wang, Hui-Min David

2016-01-01

Among many antioxidants that are used for the repairing of oxidative stress induced skin damages, we identified the enriched astaxanthin extract (EAE) from Haematococcus pluvialis as a viable ingredient. EAE was extracted from the red microalgae through supercritical fluid carbon dioxide extraction. To compare the effectiveness, EAE wastreated on human dermal fibroblasts with other components, phorbol 12-myristate 13-acetate (PMA), and doxycycline. With sirius red staining and quantitative real-time polymerase chain reaction (qRT-PCR), we found that PMA decreased the collagen concentration and production while overall the addition of doxycycline and EAE increased the collagen concentration in a trial experiments. EAE increased collagen contents through inhibited MMP1 and MMP3 mRNA expression and induced TIMP1, the antagonists of MMPs protein, gene expression. As for when tested for various proteins through western blotting, it was seen that the addition of EAE increased the expression of certain proteins that promote cell proliferation. Testing those previous solutions using growth factor assay, it was noticeable that EAE had a positive impact on cell proliferation and vascular endothelial growth factor (VEGF) than doxycycline, indicating that it was a better alternative treatment for collagen production. To sum up, the data confirmed the possible applications as medical cosmetology agentsand food supplements. PMID:27322248

In vivo hyperspectral CARS and FWM microscopy of carotenoid accumulation in H. Pluvialis

NASA Astrophysics Data System (ADS)

Slepkov, Aaron D.; Barlow, Aaron M.; Ridsdale, Andrew; McGinn, Patrick J.; Stolow, Albert

2014-02-01

Coherent anti-Stokes Raman scattering (CARS) and four-wave-mixing (FWM) microscopy are a related pair of powerful nonlinear optical characterization tools. These techniques often yield strong signals from concentrated samples, but because of their quadratic dependence on concentration, they are not typically employed for imaging or identifying dilute cellular constituents. We report here that, depending on the excitation wavelengths employed, both CARS and degenerate-FWM signals from carotenoid accumulations in alga cysts can be exceptionally large, allowing for low-power imaging of astaxanthin (AXN) deposits in Haematococcus pluvialis microalga. By use of a broadband laser pulse scheme for CARS and FWM, we are able to simultaneously collect strong intrinsic two-photon-excitation fluorescence signals from cellular chlorophyll in vivo. We show that CARS signals from astaxanthin (AXN) samples in vitro strictly follow the expected quadratic dependence on concentration, and we demonstrate the collection of wellresolved CARS spectra in the fingerprint region with sensitivity below 2mM. We suggest that multimodal nonlinear optical microscopy is sufficiently sensitive to AXN and chlorophyll concentrations that it will allow for non-invasive monitoring of carotenogenesis in live H. pluvialis microalgae.

[Protective effects of astaxanthin against oxidative damage induced by 60Co gamma-ray irradiation].

PubMed

Zhao, Wei; Jing, Xuejun; Chen, Chen; Cui, Jie; Yang, Mo; Zhang, Zunzhen

2011-09-01

To investigate the protection effect of haematococcus pluvialis (containing astaxanthin) against the impairment of anti-oxidative system and DNA damage in mice induced by 60Co gamma-rays. Fifty mice were randomly divided into five groups, i.e. three haematococcus pluvialis groups (41.7, 83.3 and 166.7 mg/kg in vegetable oil, respectively), control group and model group (vegetable oil only). All mice except control group were irradiated by 8 Gy 60Co gamma-rays 30 days later, and executed in the 4th day after irradiation. Liver cells were collected for the analysis of the integrity of DNA by comet assay, as well as MDA contents, SOD and GSH-Px activities in liver by commercial kits. Peripheral granulocyte and bone marrow nucleated cells were counted by hematocyte counter. MDA contents of model group were higher than those of control group (P < 0.01), and SOD, GSH-Px activities of model group were lower than those of control group (P < 0.01). Compared with the model group, MDA contents were decreased (P < 0.01), and SOD and GSH-Px activities were increased (P < 0.01) in all haematococcus pluvialis groups, especially in the high haematococcus pluvialis group, and the more haematococcus pluvialis in the diet of mice, the lower rate of comet tail and OTM value were shown (P < 0.01). Furthermore, the counts of peripheral granulocyte and bone marrow nucleated cells of model group were lower than those of the control group, while the counts of peripheral granulocyte and bone marrow nucleated cells of medium and high haematococcus pluvialis groups were increased significantly when compared with the model group (P < 0.01). Astaxanthin might have some protective effect against oxidative impairment and DNA damage induced by 60Co gamma-rays in mice.

Antioxidant potential of Haematococcus pluvialis extract rich in astaxanthin on colour and oxidative stability of raw ground pork meat during refrigerated storage.

PubMed

Pogorzelska, Ewelina; Godziszewska, Jolanta; Brodowska, Marta; Wierzbicka, Agnieszka

2018-01-01

Astaxanthin is proven to be one of the most potent, naturally occurring antioxidants. A rich source of astaxanthin is algae Haematoccocus pluvialis (H. pluvialis). The aim of the study was to investigate antioxidant effect of H. pluvalis extract added in different levels (0.15, 0.3 or 0.45g/kg of meat) on colour and oxidative stability of raw ground pork meat during refrigerated storage (7days). Obtained data revealed that DPPH scavenging activity of the extract at the concentration of 0.45g/kg of meat was as high as 85%. Moreover, application of higher extract doses (0.3 and 0.45g/kg) delayed lipids oxidation (lower TBARS value than control) and improved colour stability (increased a* colour parameter). Additionally, usage of 0.3 and 0.45g/kg had a positive effect on meat acceptance declared by consumers' at the final day of storage. However, the extract of H. pluvialis had no antimicrobial or antioxidative activity against myoglobin oxidation. Copyright © 2017 Elsevier Ltd. All rights reserved.

21 CFR 73.185 - Haematococcus algae meal.

Code of Federal Regulations, 2012 CFR

2012-04-01

... 21 Food and Drugs 1 2012-04-01 2012-04-01 false Haematococcus algae meal. 73.185 Section 73.185... COLOR ADDITIVES EXEMPT FROM CERTIFICATION Foods § 73.185 Haematococcus algae meal. (a) Identity. (1) The color additive haematococcus algae meal consists of the comminuted and dried cells of the alga...

21 CFR 73.185 - Haematococcus algae meal.

Code of Federal Regulations, 2014 CFR

2014-04-01

... 21 Food and Drugs 1 2014-04-01 2014-04-01 false Haematococcus algae meal. 73.185 Section 73.185... COLOR ADDITIVES EXEMPT FROM CERTIFICATION Foods § 73.185 Haematococcus algae meal. (a) Identity. (1) The color additive haematococcus algae meal consists of the comminuted and dried cells of the alga...

21 CFR 73.185 - Haematococcus algae meal.

Code of Federal Regulations, 2010 CFR

2010-04-01

... 21 Food and Drugs 1 2010-04-01 2010-04-01 false Haematococcus algae meal. 73.185 Section 73.185... COLOR ADDITIVES EXEMPT FROM CERTIFICATION Foods § 73.185 Haematococcus algae meal. (a) Identity. (1) The color additive haematococcus algae meal consists of the comminuted and dried cells of the alga...

21 CFR 73.185 - Haematococcus algae meal.

Code of Federal Regulations, 2011 CFR

2011-04-01

... 21 Food and Drugs 1 2011-04-01 2011-04-01 false Haematococcus algae meal. 73.185 Section 73.185... COLOR ADDITIVES EXEMPT FROM CERTIFICATION Foods § 73.185 Haematococcus algae meal. (a) Identity. (1) The color additive haematococcus algae meal consists of the comminuted and dried cells of the alga...

21 CFR 73.185 - Haematococcus algae meal.

Code of Federal Regulations, 2013 CFR

2013-04-01

... 21 Food and Drugs 1 2013-04-01 2013-04-01 false Haematococcus algae meal. 73.185 Section 73.185... COLOR ADDITIVES EXEMPT FROM CERTIFICATION Foods § 73.185 Haematococcus algae meal. (a) Identity. (1) The color additive haematococcus algae meal consists of the comminuted and dried cells of the alga...

Metabolite profiling and integrative modeling reveal metabolic constraints for carbon partitioning under nitrogen starvation in the green algae Haematococcus pluvialis.

PubMed

Recht, Lee; Töpfer, Nadine; Batushansky, Albert; Sikron, Noga; Gibon, Yves; Fait, Aaron; Nikoloski, Zoran; Boussiba, Sammy; Zarka, Aliza

2014-10-31

The green alga Hematococcus pluvialis accumulates large amounts of the antioxidant astaxanthin under inductive stress conditions, such as nitrogen starvation. The response to nitrogen starvation and high light leads to the accumulation of carbohydrates and fatty acids as well as increased activity of the tricarboxylic acid cycle. Although the behavior of individual pathways has been well investigated, little is known about the systemic effects of the stress response mechanism. Here we present time-resolved metabolite, enzyme activity, and physiological data that capture the metabolic response of H. pluvialis under nitrogen starvation and high light. The data were integrated into a putative genome-scale model of the green alga to in silico test hypotheses of underlying carbon partitioning. The model-based hypothesis testing reinforces the involvement of starch degradation to support fatty acid synthesis in the later stages of the stress response. In addition, our findings support a possible mechanism for the involvement of the increased activity of the tricarboxylic acid cycle in carbon repartitioning. Finally, the in vitro experiments and the in silico modeling presented here emphasize the predictive power of large scale integrative approaches to pinpoint metabolic adjustment to changing environments. © 2014 by The American Society for Biochemistry and Molecular Biology, Inc.

Accurate quantification of astaxanthin from Haematococcus crude extract spectrophotometrically

NASA Astrophysics Data System (ADS)

Li, Yeguang; Miao, Fengping; Geng, Yahong; Lu, Dayan; Zhang, Chengwu; Zeng, Mingtao

2012-07-01

The influence of alkali on astaxanthin and the optimal working wave length for measurement of astaxanthin from Haematococcus crude extract were investigated, and a spectrophotometric method for precise quantification of the astaxanthin based on the method of Boussiba et al. was established. According to Boussiba's method, alkali treatment destroys chlorophyll. However, we found that: 1) carotenoid content declined for about 25% in Haematococcus fresh cysts and up to 30% in dry powder of Haematococcus broken cysts after alkali treatment; and 2) dimethyl sulfoxide (DMSO)-extracted chlorophyll of green Haematococcus bares little absorption at 520-550 nm. Interestingly, a good linear relationship existed between absorbance at 530 nm and astaxanthin content, while an unknown interference at 540-550 nm was detected in our study. Therefore, with 530 nm as working wavelength, the alkali treatment to destroy chlorophyll was not necessary and the influence of chlorophyll, other carotenoids, and the unknown interference could be avoided. The astaxanthin contents of two samples were measured at 492 nm and 530 nm; the measured values at 530 nm were 2.617 g/100 g and 1.811 g/100 g. When compared with the measured values at 492 nm, the measured values at 530 nm decreased by 6.93% and 11.96%, respectively. The measured values at 530 nm are closer to the true astaxanthin contents in the samples. The data show that 530 nm is the most suitable wave length for spectrophotometric determination to the astaxanthin in Haematococcus crude extract.

Characterization of microalgal carotenoids by mass spectrometry and their bioavailability and antioxidant properties elucidated in rat model.

PubMed

Ranga Rao, A; Raghunath Reddy, R L; Baskaran, V; Sarada, R; Ravishankar, G A

2010-08-11

Of the total carotenoids in respective algal samples, beta-carotene in Spirulina platensis was 69.5%, astaxanthin and its esters in Haematococcus pluvialis was 81.38%, and lutein in Botryococcus braunii was 74.6%. The carotenoids were characterized by mass spectrometry. A time-course study of carotenoids in rats after administration of microalgal biomass showed peak levels in plasma, liver, and eyes at 2, 4, and 6 h, respectively. Beta-carotene accumulation in Spirulina-fed rats was maximum in eye tissues at 6 h. Similarly, levels of astaxanthin and lutein in Haematococcus- and Botryococcus-fed rats were also maximal in eye tissues. Astaxanthin from H. pluvialis showed better bioavailability than beta-carotene and lutein. The antioxidant enzymes, catalase, superoxide dismutase, peroxidase, and TBARS were significantly high in plasma at 2 h and in liver at 4 h, evidently offering protection from free radicals. This study implies that microalgae can be a good source of carotenoids of high bioavailability and nutraceutical value.

Growth and development time of subtropical Cladocera Diaphanosoma birgei Korinek, 1981 fed with different microalgal diets.

PubMed

Sipaúba-Tavares, L H; Truzzi, B S; Berchielli-Morais, F A

2014-05-01

The aim of this work was to investigate the growth performance of Diaphanosoma birgei fed with two Chlorophyceae algae, Ankistrodesmus gracilis and Haematococcus pluvialis using monoalgal diets and simpler mixed diets. D. birgei was daily fed on four treatments: 1) 100% Ankistrodesmus gracilis (Ag); 2) 100% Haematoccocus pluvialis (Hp); 3) 25% A. gracilis + 75% H. pluvialis (Ag-25+Hp-75) and 4) 75% A. gracilis + 25% H. pluvialis (Ag-75+Hp-25). The fecundity curve of D. birgei showed that the mixed feed Ag-25+Hp-75 and temperature 24±2°C triggered fast fecundity at approximately two days. The fecundity was low when based only on H. pluvialis (Hp), albeit with greater longevity (19 days) and a higher number of broods (8). D. birgei fed on Ag and Ag-75+Hp-25 diets in this experiment sustained higher growth rate and higher lipid content in these treatments. The present study showed that A. gracilis diet and mixed microalgae diets tested were able to support the egg production and development of D. birgei.

Cell-wall disruption and lipid/astaxanthin extraction from microalgae: Chlorella and Haematococcus.

PubMed

Kim, Dong-Yeon; Vijayan, Durairaj; Praveenkumar, Ramasamy; Han, Jong-In; Lee, Kyubock; Park, Ji-Yeon; Chang, Won-Seok; Lee, Jin-Suk; Oh, You-Kwan

2016-01-01

Recently, biofuels and nutraceuticals produced from microalgae have emerged as major interests, resulting in intensive research of the microalgal biorefinery process. In this paper, recent developments in cell-wall disruption and extraction methods are reviewed, focusing on lipid and astaxanthin production from the biotechnologically important microalgae Chlorella and Haematococcus, respectively. As a common, critical bottleneck for recovery of intracellular components such as lipid and astaxanthin from these microalgae, the composition and structure of rigid, thick cell-walls were analyzed. Various chemical, physical, physico-chemical, and biological methods applied for cell-wall breakage and lipid/astaxanthin extraction from Chlorella and Haematococcus are discussed in detail and compared based on efficiency, energy consumption, type and dosage of solvent, biomass concentration and status (wet/dried), toxicity, scalability, and synergistic combinations. This report could serve as a useful guide to the implementation of practical downstream processes for recovery of valuable products from microalgae including Chlorella and Haematococcus. Copyright © 2015 Elsevier Ltd. All rights reserved.

Influence of TiO2 Nanoparticles on Growth and Phenolic Compounds Production in Photosynthetic Microorganisms

PubMed Central

Comotto, Mattia; Casazza, Alessandro Alberto; Aliakbarian, Bahar; Caratto, Valentina; Ferretti, Maurizio; Perego, Patrizia

2014-01-01

The influence of titanium dioxide nanoparticles (pure anatase and 15% N doped anatase) on the growth of Chlorella vulgaris, Haematococcus pluvialis, and Arthrospira platensis was investigated. Results showed that pure anatase can lead to a significant growth inhibition of C. vulgaris and A. platensis (17.0 and 74.1%, resp.), while for H. pluvialis the nanoparticles do not cause a significant inhibition. Since in these stress conditions photosynthetic microorganisms can produce antioxidant compounds in order to prevent cell damages, we evaluated the polyphenols content either inside the cells or released in the medium. Although results did not show a significant difference in C. vulgaris, the phenolic concentrations of two other microorganisms were statistically affected by the presence of titanium dioxide. In particular, 15% N doped anatase resulted in a higher production of extracellular antioxidant compounds, reaching the concentration of 65.2 and 68.0 mg gDB −1 for H. pluvialis and A. platensis, respectively. PMID:25610914

Phytophthora pluvialis, a new species from mixed tanoak-Douglas-fir forests of western Oregon, U.S.A

Treesearch

Paul Reeser; Wendy Sutton; Everett Hansen

2013-01-01

A new species, Phytophthora pluvialis is described. P. pluvialis has been recovered from streams, soil and canopy drip in the mixed tanoak-Douglas-fir forest in Curry County, Oregon, and in two additional streams in other areas of western Oregon. It has been found only rarely in association with twig and stem cankers on tanoak...

A Novel Adaptation Mechanism Underpinning Algal Colonization of a Nuclear Fuel Storage Pond.

PubMed

MeGraw, Victoria E; Brown, Ashley R; Boothman, Christopher; Goodacre, Royston; Morris, Katherine; Sigee, David; Anderson, Lizzie; Lloyd, Jonathan R

2018-06-26

Geochemical analyses alongside molecular techniques were used to characterize the microbial ecology and biogeochemistry of an outdoor spent nuclear fuel storage pond at Sellafield, United Kingdom, that is susceptible to seasonal algal blooms that cause plant downtime. 18S rRNA gene profiling of the filtered biomass samples showed the increasing dominance of a species closely related to the alga Haematococcus pluvialis , alongside 16S rRNA genes affiliated with a diversity of freshwater bacteria, including Proteobacteria and Cyanobacteria High retention of 137 Cs and 90 Sr on pond water filters coincided with high levels of microbial biomass in the pond, suggesting that microbial colonization may have an important control on radionuclide fate in the pond. To interpret the unexpected dominance of Haematococcus species during bloom events in this extreme environment, the physiological response of H. pluvialis to environmentally relevant ionizing radiation doses was assessed. Irradiated laboratory cultures produced significant quantities of the antioxidant astaxanthin, consistent with pigmentation observed in pond samples. Fourier transform infrared (FT-IR) spectroscopy suggested that radiation did not have a widespread impact on the metabolic fingerprint of H. pluvialis in laboratory experiments, despite the 80-Gy dose. This study suggests that the production of astaxanthin-rich encysted cells may be related to the preservation of the Haematococcus phenotype, potentially allowing it to survive oxidative stress arising from radiation doses associated with the spent nuclear fuel. The oligotrophic and radiologically extreme conditions in this environment do not prevent extensive colonization by microbial communities, which play a defining role in controlling the biogeochemical fate of major radioactive species present. IMPORTANCE Spent nuclear fuel is stored underwater in large ponds prior to processing and disposal. Such environments are intensively radioactive but

Raman-Activated Droplet Sorting (RADS) for Label-Free High-Throughput Screening of Microalgal Single-Cells.

PubMed

Wang, Xixian; Ren, Lihui; Su, Yetian; Ji, Yuetong; Liu, Yaoping; Li, Chunyu; Li, Xunrong; Zhang, Yi; Wang, Wei; Hu, Qiang; Han, Danxiang; Xu, Jian; Ma, Bo

2017-11-21

Raman-activated cell sorting (RACS) has attracted increasing interest, yet throughput remains one major factor limiting its broader application. Here we present an integrated Raman-activated droplet sorting (RADS) microfluidic system for functional screening of live cells in a label-free and high-throughput manner, by employing AXT-synthetic industrial microalga Haematococcus pluvialis (H. pluvialis) as a model. Raman microspectroscopy analysis of individual cells is carried out prior to their microdroplet encapsulation, which is then directly coupled to DEP-based droplet sorting. To validate the system, H. pluvialis cells containing different levels of AXT were mixed and underwent RADS. Those AXT-hyperproducing cells were sorted with an accuracy of 98.3%, an enrichment ratio of eight folds, and a throughput of ∼260 cells/min. Of the RADS-sorted cells, 92.7% remained alive and able to proliferate, which is equivalent to the unsorted cells. Thus, the RADS achieves a much higher throughput than existing RACS systems, preserves the vitality of cells, and facilitates seamless coupling with downstream manipulations such as single-cell sequencing and cultivation.

Conversion of stranded waste-stream carbon and nutrients into value-added products via metabolically coupled binary heterotroph-photoautotroph system.

PubMed

Bohutskyi, Pavlo; Kucek, Leo A; Hill, Eric; Pinchuk, Grigoriy E; Mundree, Sagadevan G; Beliaev, Alexander S

2018-07-01

Growth of heterotrophic bacterium Bacillus subtilis was metabolically coupled with the photosynthetic activity of an astaxanthin-producing alga Haematococcus pluvialis for conversion of starch-containing waste stream into carotenoid-enriched biomass. The H. pluvialis accounted for 63% of the produced co-culture biomass of 2.2 g/L. Importantly, the binary system requires neither exogenous supply of gaseous substrates nor application of energy-intensive mass transfer technologies due to in-situ exchange in CO 2 and O 2 . The maximum reduction in COD, total nitrogen and phosphorus reached 65%, 55% and 30%, respectively. Conducted techno-economic assessment suggested that the astaxanthin-rich biomass may potentially offset the costs of waste treatment, and, with specific productivity enhancements (induction of astaxanthin to 2% and increase H. pluvialis fraction to 80%), provide and additional revenue stream. The outcome of this study demonstrates a successful proof-of-principle for conversion of waste carbon and nutrients into value-added products through metabolic coupling of heterotrophic and phototrophic metabolisms. Copyright © 2018. Published by Elsevier Ltd.

Astaxanthin: Sources, Extraction, Stability, Biological Activities and Its Commercial Applications—A Review

PubMed Central

Ambati, Ranga Rao; Siew Moi, Phang; Ravi, Sarada; Aswathanarayana, Ravishankar Gokare

2014-01-01

There is currently much interest in biological active compounds derived from natural resources, especially compounds that can efficiently act on molecular targets, which are involved in various diseases. Astaxanthin (3,3′-dihydroxy-β, β′-carotene-4,4′-dione) is a xanthophyll carotenoid, contained in Haematococcus pluvialis, Chlorella zofingiensis, Chlorococcum, and Phaffia rhodozyma. It accumulates up to 3.8% on the dry weight basis in H. pluvialis. Our recent published data on astaxanthin extraction, analysis, stability studies, and its biological activities results were added to this review paper. Based on our results and current literature, astaxanthin showed potential biological activity in in vitro and in vivo models. These studies emphasize the influence of astaxanthin and its beneficial effects on the metabolism in animals and humans. Bioavailability of astaxanthin in animals was enhanced after feeding Haematococcus biomass as a source of astaxanthin. Astaxanthin, used as a nutritional supplement, antioxidant and anticancer agent, prevents diabetes, cardiovascular diseases, and neurodegenerative disorders, and also stimulates immunization. Astaxanthin products are used for commercial applications in the dosage forms as tablets, capsules, syrups, oils, soft gels, creams, biomass and granulated powders. Astaxanthin patent applications are available in food, feed and nutraceutical applications. The current review provides up-to-date information on astaxanthin sources, extraction, analysis, stability, biological activities, health benefits and special attention paid to its commercial applications. PMID:24402174

Chlorella zofingiensis as an Alternative Microalgal Producer of Astaxanthin: Biology and Industrial Potential

PubMed Central

Liu, Jin; Sun, Zheng; Gerken, Henri; Liu, Zheng; Jiang, Yue; Chen, Feng

2014-01-01

Astaxanthin (3,3′-dihydroxy-β,β-carotene-4,4′-dione), a high-value ketocarotenoid with a broad range of applications in food, feed, nutraceutical, and pharmaceutical industries, has been gaining great attention from science and the public in recent years. The green microalgae Haematococcus pluvialis and Chlorella zofingiensis represent the most promising producers of natural astaxanthin. Although H. pluvialis possesses the highest intracellular astaxanthin content and is now believed to be a good producer of astaxanthin, it has intrinsic shortcomings such as slow growth rate, low biomass yield, and a high light requirement. In contrast, C. zofingiensis grows fast phototrophically, heterotrophically and mixtrophically, is easy to be cultured and scaled up both indoors and outdoors, and can achieve ultrahigh cell densities. These robust biotechnological traits provide C. zofingiensis with high potential to be a better organism than H. pluvialis for mass astaxanthin production. This review aims to provide an overview of the biology and industrial potential of C. zofingiensis as an alternative astaxanthin producer. The path forward for further expansion of the astaxanthin production from C. zofingiensis with respect to both challenges and opportunities is also discussed. PMID:24918452

Highly efficient biosynthesis of astaxanthin in Saccharomyces cerevisiae by integration and tuning of algal crtZ and bkt.

PubMed

Zhou, Pingping; Ye, Lidan; Xie, Wenping; Lv, Xiaomei; Yu, Hongwei

2015-10-01

Astaxanthin is a highly valued carotenoid with strong antioxidant activity and has wide applications in aquaculture, food, cosmetic, and pharmaceutical industries. The market demand for natural astaxanthin promotes research in metabolic engineering of heterologous hosts for astaxanthin production. In this study, an astaxanthin-producing Saccharomyces cerevisiae strain was created by successively introducing the Haematococcus pluvialis β-carotenoid hydroxylase (crtZ) and ketolase (bkt) genes into a previously constructed β-carotene hyperproducer. Further integration of strategies including codon optimization, gene copy number adjustment, and iron cofactor supplementation led to significant increase in the astaxanthin production, reaching up to 4.7 mg/g DCW in the shake-flask cultures which is the highest astaxanthin content in S. cerevisiae reported to date. Besides, the substrate specificity of H. pluvialis CrtZ and BKT and the probable formation route of astaxanthin from β-carotene in S. cerevisiae were figured out by expressing the genes separately and in combination. The yeast strains engineered in this work provide a basis for further improving biotechnological production of astaxanthin and might offer a useful general approach to the construction of heterologous biosynthetic pathways for other natural products.

Optimization of protein electroextraction from microalgae by a flow process.

PubMed

Coustets, Mathilde; Joubert-Durigneux, Vanessa; Hérault, Josiane; Schoefs, Benoît; Blanckaert, Vincent; Garnier, Jean-Pierre; Teissié, Justin

2015-06-01

Classical methods, used for large scale treatments such as mechanical or chemical extractions, affect the integrity of extracted cytosolic protein by releasing proteases contained in vacuoles. Our previous experiments on flow processes electroextraction on yeasts proved that pulsed electric field technology allows preserving the integrity of released cytosolic proteins, by not affecting vacuole membranes. Furthermore, large cell culture volumes are easily treated by the flow technology. Based on this previous knowledge, we developed a new protocol in order to electro-extract total cytoplasmic proteins from microalgae (Nannochloropsis salina, Chlorella vulgaris and Haematococcus pluvialis). Given that induction of electropermeabilization is under the control of target cell size, as the mean diameter for N. salina is only 2.5 μm, we used repetitive 2 ms long pulses of alternating polarities with stronger field strengths than previously described for yeasts. The electric treatment was followed by a 24h incubation period in a salty buffer. The amount of total protein release was observed by a classical Bradford assay. A more accurate evaluation of protein release was obtained by SDS-PAGE. Similar results were obtained with C. vulgaris and H. pluvialis under milder electrical conditions as expected from their larger size. Copyright © 2014 Elsevier B.V. All rights reserved.

Enhancing Photon Utilization Efficiency for Astaxanthin Production from Haematococcus lacustris Using a Split-Column Photobioreactor.

PubMed

Kim, Z-Hun; Park, Hanwool; Lee, Ho-Sang; Lee, Choul-Gyun

2016-07-28

A split-column photobioreactor (SC-PBR), consisting of two bubble columns with different sizes, was developed to enhance the photon utilization efficiency in an astaxanthin production process from Haematococcus lacustris. Among the two columns, only the smaller column of SC-PBR was illuminated. Astaxanthin productivities and photon efficiencies of the SC-PBRs were compared with a standard bubble-column PBR (BC-PBR). Astaxanthin productivity of SC-PBR was improved by 28%, and the photon utilization efficiencies were 28-366% higher than the original BC-PBR. The results clearly show that the effective light regime of SC-PBR could enhance the production of astaxanthin.

Astaxanthin preparation by fermentation of esters from Haematococcus pluvialis algal extracts with Stenotrophomonas species.

PubMed

Dong, Hao; Li, Xuemin; Xue, Changhu; Mao, Xiangzhao

2016-05-01

Natural astaxanthin (Ax) is an additive that is widely used because of its beneficial biochemical functions. However, the methods used to produce free Ax have drawbacks. Chemical saponification methods produce several by-products, and lipase-catalyzed hydrolysis methods are not cost effective. In this study, a bacterial strain of Stenotrophomonas sp. was selected to enzymatically catalyze the saponification of Ax esters to produce free all-trans-Ax. Through single-factor experiments and a Box-Behnken design, the optimal fermentation conditions were determined as follows: a seed culture age of 37.79 h, an inoculum concentration of 5.92%, and an initial broth pH of 6.80. Under these conditions, a fermentation curve was drawn, and the optimal fermentation time was shown to be 60 h. At 60 h, the degradation rate of the Ax esters was 98.08%, and the yield of free all-trans-Ax was 50.130 μg/mL. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:649-656, 2016. © 2016 American Institute of Chemical Engineers.

Bioavailability of astaxanthin in Haematococcus algal extract: the effects of timing of diet and smoking habits.

PubMed

Okada, Yumika; Ishikura, Masaharu; Maoka, Takashi

2009-09-01

Astaxanthin is a caroteonid that possesses strong antioxidant activity. Recently, many studies on biological activity have been reported. In general, the absorption of carotenoids is affected greatly by diet and by smoking. In this report, we investigated astaxanthin pharmacokinetics after administration of Haematococcus algal extract, a source of astaxanthin, to smokers and nonsmokers before and after a meal; astaxanthin was given before the meal to nonsmokers (n = 7), after the meal to nonsmokers (n = 6), and after the meal to smokers (n = 7), then serum samples were analyzed. The timing of administration greatly affected astaxanthin bioavailability including the area under the curve (AUC(0-168), 2,968 + or - 959 microg h/l in the before-meal group vs. 7,219 + or - 3,118 microg h/l in the after-meal group), indicating high availability in the after-meal group. Smoking also affected the pharmacokinetic parameters and reduced the half-life (t(1/2)) of astaxanthin elimination significantly.

A Simple Method to Decode the Complete 18-5.8-28S rRNA Repeated Units of Green Algae by Genome Skimming.

PubMed

Lin, Geng-Ming; Lai, Yu-Heng; Audira, Gilbert; Hsiao, Chung-Der

2017-11-06

Green algae, Chlorella ellipsoidea , Haematococcus pluvialis and Aegagropila linnaei (Phylum Chlorophyta) were simultaneously decoded by a genomic skimming approach within 18-5.8-28S rRNA region. Whole genomic DNAs were isolated from green algae and directly subjected to low coverage genome skimming sequencing. After de novo assembly and mapping, the size of complete 18-5.8-28S rRNA repeated units for three green algae were ranged from 5785 to 6028 bp, which showed high nucleotide diversity (π is around 0.5-0.6) within ITS1 and ITS2 (Internal Transcribed Spacer) regions. Previously, the evolutional diversity of algae has been difficult to decode due to the inability design universal primers that amplify specific marker genes across diverse algal species. In this study, our method provided a rapid and universal approach to decode the 18-5.8-28S rRNA repeat unit in three green algal species. In addition, the completely sequenced 18-5.8-28S rRNA repeated units provided a solid nuclear marker for phylogenetic and evolutionary analysis for green algae for the first time.

Solar spectral conversion for improving the photosynthetic activity in algae reactors.

PubMed

Wondraczek, Lothar; Batentschuk, Miroslaw; Schmidt, Markus A; Borchardt, Rudolf; Scheiner, Simon; Seemann, Benjamin; Schweizer, Peter; Brabec, Christoph J

2013-01-01

Sustainable biomass production is expected to be one of the major supporting pillars for future energy supply, as well as for renewable material provision. Algal beds represent an exciting resource for biomass/biofuel, fine chemicals and CO2 storage. Similar to other solar energy harvesting techniques, the efficiency of algal photosynthesis depends on the spectral overlap between solar irradiation and chloroplast absorption. Here we demonstrate that spectral conversion can be employed to significantly improve biomass growth and oxygen production rate in closed-cycle algae reactors. For this purpose, we adapt a photoluminescent phosphor of the type Ca0.59Sr0.40Eu0.01S, which enables efficient conversion of the green part of the incoming spectrum into red light to better match the Qy peak of chlorophyll b. Integration of a Ca0.59Sr0.40Eu0.01S backlight converter into a flat panel algae reactor filled with Haematococcus pluvialis as a model species results in significantly increased photosynthetic activity and algae reproduction rate.

Production of stable food-grade microencapsulated astaxanthin by vibrating nozzle technology.

PubMed

Vakarelova, Martina; Zanoni, Francesca; Lardo, Piergiovanni; Rossin, Giacomo; Mainente, Federica; Chignola, Roberto; Menin, Alessia; Rizzi, Corrado; Zoccatelli, Gianni

2017-04-15

Astaxanthin is a carotenoid known for its strong antioxidant and health-promoting characteristics, but it is also highly degradable and thus unsuited for several applications. We developed a sustainable method for the extraction and the production of stable astaxanthin microencapsulates. Nearly 2% astaxanthin was extracted by high-pressure homogenization of dried Haematococcus pluvialis cells in soybean oil. Astaxanthin-enriched oil was encapsulated in alginate and low-methoxyl pectin by Ca 2+ -mediated vibrating-nozzle extrusion technology. The 3% pectin microbeads resulted the best compromise between sphericity and oil retention upon drying. We monitored the stability of these astaxanthin beads under four different conditions of light, temperature and oxygen exposition. After 52weeks, the microbeads showed a total-astaxanthin retention of 94.1±4.1% (+4°C/-light/+O 2 ), 83.1±3.2% (RT/-light/-O 2 ), 38.3±2.2% (RT/-light/+O2), and 57.0±0.4% (RT/+light/+O 2 ), with different degradation kinetics. Refrigeration, therefore, resulted the optimal storage condition to preserve astaxanthin stability. Copyright © 2016 Elsevier Ltd. All rights reserved.

Colonization of Snow by Microorganisms as Revealed Using Miniature Raman Spectrometers - Possibilities for Detecting Carotenoids of Psychrophiles on Mars?

NASA Astrophysics Data System (ADS)

Jehlička, Jan; Culka, Adam; Nedbalová, Linda

2016-12-01

We tested the potential of a miniaturized Raman spectrometer for use in field detection of snow algae pigments. A miniature Raman spectrometer, equipped with an excitation laser at 532 nm, allowed for the detection of carotenoids in cells of Chloromonas nivalis and Chlamydomonas nivalis at different stages of their life cycle. Astaxanthin, the major photoprotective pigment, was detected in algal blooms originating in snows at two alpine European sites that differed in altitude (Krkonoše Mts., Czech Republic, 1502 m a.s.l., and Ötztal Alps, Austria, 2790 m a.s.l.). Comparison is made with a common microalga exclusively producing astaxanthin (Haematococcus pluvialis). The handheld Raman spectrometer is a useful tool for fast and direct field estimations of the presence of carotenoids (mainly astaxanthin) within blooms of snow algae. Application of miniature Raman instruments as well as flight prototypes in areas where microbes are surviving under extreme conditions is an important stage in preparation for successful deployment of this kind of instrumentation in the framework of forthcoming astrobiological missions to Mars.

Effects of non-steroidal anti-inflammatory drugs on cyanobacteria and algae in laboratory strains and in natural algal assemblages.

PubMed

Bácsi, István; B-Béres, Viktória; Kókai, Zsuzsanna; Gonda, Sándor; Novák, Zoltán; Nagy, Sándor Alex; Vasas, Gábor

2016-05-01

In recent years measurable concentrations of non-steroidal anti-inflammatory drugs (NSAIDs) have been shown in the aquatic environment as a result of increasing human consumption. Effects of five frequently used non-steroidal anti-inflammatory drugs (diclofenac, diflunisal, ibuprofen, mefenamic acid and piroxicam in 0.1 mg ml(-1) concentration) in batch cultures of cyanobacteria (Synechococcus elongatus, Microcystis aeruginosa, Cylindrospermopsis raciborskii), and eukaryotic algae (Desmodesmus communis, Haematococcus pluvialis, Cryptomonas ovata) were studied. Furthermore, the effects of the same concentrations of NSAIDs were investigated in natural algal assemblages in microcosms. According to the changes of chlorophyll-a content, unicellular cyanobacteria seemed to be more tolerant to NSAIDs than eukaryotic algae in laboratory experiments. Growth of eukaryotic algae was reduced by all drugs, the cryptomonad C. ovata was the most sensitive to NSAIDs, while the flagellated green alga H. pluvialis was more sensitive than the non-motile green alga D. communis. NSAID treatments had weaker impact in the natural assemblages dominated by cyanobacteria than in the ones dominated by eukaryotic algae, confirming the results of laboratory experiments. Diversity and number of functional groups did not change notably in cyanobacteria dominated assemblages, while they decreased significantly in eukaryotic algae dominated ones compared to controls. The results highlight that cyanobacteria (especially unicellular ones) are less sensitive to the studied, mostly hardly degradable NSAIDs, which suggest that their accumulation in water bodies may contribute to the expansion of cyanobacterial mass productions in appropriate environmental circumstances by pushing back eukaryotic algae. Thus, these contaminants require special attention during wastewater treatment and monitoring of surface waters. Copyright © 2016 Elsevier Ltd. All rights reserved.

Effect of Diets Supplemented with Different Sources of Astaxanthin on the Gonad of the Sea Urchin Anthocidaris crassispina

PubMed Central

Peng, Juan; Yuan, Jian-Ping; Wang, Jiang-Hai

2012-01-01

The effect of the microalgae Haematococcus pluvialis and Chorella zofingiensis, and synthetic astaxanthin on the gonad of the sea urchin Anthocidaris crassispina was studied. The basal diet was supplemented with H. pluvialis, C. zofingiensis, or synthetic astaxanthin, at two levels of astaxanthin (approximately 400 mg/kg and 100 mg/kg), to obtain the experimental diets HP1, HP2, CZ1, CZ2, AST1, and AST2, respectively, for two months of feeding experiment. The results showed that the concentrations of astaxanthin in the gonads of the sea urchins fed these experimental diets ranged from 0.15 to 3.01 mg/kg dry gonad weight. The higher astaxanthin levels (>2.90 mg/kg) were found in the gonads of the sea urchins fed the diets HP1 (containing 380 mg/kg of astaxanthins, mostly mono- and diesters) and AST1 (containing 385 mg/kg of synthetic astaxanthin). The lowest astaxanthin level (0.15 mg/kg) was detected in the gonads of the sea urchins fed the diet CZ2 (containing 98 mg/kg of astaxanthins, mostly diesters). Furthermore, the highest canthaxanthin level (7.48 mg/kg) was found in the gonads of the sea urchins fed the diet CZ1 (containing 387 mg/kg of astaxanthins and 142 mg/kg of canthaxanthin), suggesting that astaxanthins, especially astaxanthin esters, might not be assimilated as easily as canthaxanthin by the sea urchins. Our results show that sea urchins fed diets containing astaxanthin pigments show higher incorporation of these known antioxidant constituents, with the resultant seafood products therefore being of potential higher nutritive value. PMID:23016124

Specific light uptake rates can enhance astaxanthin productivity in Haematococcus lacustris.

PubMed

Lee, Ho-Sang; Kim, Z-Hun; Park, Hanwool; Lee, Choul-Gyun

2016-05-01

Lumostatic operation was applied for efficient astaxanthin production in autotrophic Haematococcus lacustris cultures using 0.4-L bubble column photobioreactors. The lumostatic operation in this study was performed with three different specific light uptake rates (q(e)) based on cell concentration, cell projection area, and fresh weight as one-, two- and three-dimensional characteristics values, respectively. The q(e) value from the cell concentration (q(e1D)) obtained was 13.5 × 10⁻⁸ μE cell⁻¹ s⁻¹, and the maximum astaxanthin concentration was increased to 150 % compared to that of a control with constant light intensity. The other optimum q e values by cell projection area (q(e2D)) and fresh weight (q( e3D)) were determined to be 195 μE m⁻² s⁻¹ and 10.5 μE g⁻¹ s⁻¹ for astaxanthin production, respectively. The maximum astaxanthin production from the lumostatic cultures using the parameters controlled by cell projection area (2D) and fresh weight (3D) also increased by 36 and 22% over that of the controls, respectively. When comparing the optimal q e values among the three different types, the lumostatic cultures using q(e) based on fresh weight showed the highest astaxanthin productivity (22.8 mg L⁻¹ day⁻¹), which was a higher level than previously reported. The lumostatic operations reported here demonstrated that more efficient and effective astaxanthin production was obtained by H. lacustris than providing a constant light intensity, regardless of which parameter is used to calculate the specific light uptake rate.

Colonization of Snow by Microorganisms as Revealed Using Miniature Raman Spectrometers-Possibilities for Detecting Carotenoids of Psychrophiles on Mars?

PubMed

Jehlička, Jan; Culka, Adam; Nedbalová, Linda

2016-12-01

We tested the potential of a miniaturized Raman spectrometer for use in field detection of snow algae pigments. A miniature Raman spectrometer, equipped with an excitation laser at 532 nm, allowed for the detection of carotenoids in cells of Chloromonas nivalis and Chlamydomonas nivalis at different stages of their life cycle. Astaxanthin, the major photoprotective pigment, was detected in algal blooms originating in snows at two alpine European sites that differed in altitude (Krkonoše Mts., Czech Republic, 1502 m a.s.l., and Ötztal Alps, Austria, 2790 m a.s.l.). Comparison is made with a common microalga exclusively producing astaxanthin (Haematococcus pluvialis). The handheld Raman spectrometer is a useful tool for fast and direct field estimations of the presence of carotenoids (mainly astaxanthin) within blooms of snow algae. Application of miniature Raman instruments as well as flight prototypes in areas where microbes are surviving under extreme conditions is an important stage in preparation for successful deployment of this kind of instrumentation in the framework of forthcoming astrobiological missions to Mars. Key Words: Snow algae-Chloromonas nivalis-Chlamydomonas nivalis-On-site field detection-Raman spectroscopy-Astaxanthin. Astrobiology 16, 913-924.

Preventive effect of dietary astaxanthin on UVA-induced skin photoaging in hairless mice.

PubMed

Komatsu, Toshiyuki; Sasaki, Suguru; Manabe, Yuki; Hirata, Takashi; Sugawara, Tatsuya

2017-01-01

Astaxanthin, a carotenoid found mainly in seafood, has potential clinical applications due to its antioxidant activity. In this study, we evaluated the effect of dietary astaxanthin derived from Haematococcus pluvialis on skin photoaging in UVA-irradiated hairless mice by assessing various parameters of photoaging. After chronic ultraviolet A (UVA) exposure, a significant increase in transepidermal water loss (TEWL) and wrinkle formation in the dorsal skin caused by UVA was observed, and dietary astaxanthin significantly suppressed these photoaging features. We found that the mRNA expression of lympho-epithelial Kazal-type-related inhibitor, steroid sulfatase, and aquaporin 3 in the epidermis was significantly increased by UVA irradiation for 70 days, and dietary astaxanthin significantly suppressed these increases in mRNA expression to be comparable to control levels. In the dermis, the mRNA expression of matrix metalloprotease 13 was increased by UVA irradiation and significantly suppressed by dietary astaxanthin. In addition, HPLC-PDA analysis confirmed that dietary astaxanthin reached not only the dermis but also the epidermis. Our results indicate that dietary astaxanthin accumulates in the skin and appears to prevent the effects of UVA irradiation on filaggrin metabolism and desquamation in the epidermis and the extracellular matrix in the dermis.

Recent breakthroughs in the biology of astaxanthin accumulation by microalgal cell.

PubMed

Solovchenko, Alexei E

2015-09-01

Massive accumulation of the secondary ketokarotenoid astaxanthin is a characteristic stress response of certain microalgal species with Haematococcus pluvialis as an illustrious example. The carotenogenic response confers these organisms a remarkable ability to survive in extremely unfavorable environments and makes them the richest source of natural astaxanthin. Exerting a plethora of beneficial effects on human and animal health, astaxanthin is among the most important bioproducts from microalgae. Though our understanding of astaxanthin biosynthesis, induction, and regulation is far from complete, this gap is filling rapidly with new knowledge generated predominantly by application of advanced "omics" approaches. This review focuses on the most recent progress in the biology of astaxanthin accumulation in microalgae including the genomic, proteomic, and metabolomics insights into the induction and regulation of secondary carotenogenesis and its role in stress tolerance of the photosynthetic microorganisms. Special attention is paid to the coupling of the carotenoid and lipid biosynthesis as well as deposition of astaxanthin in the algal cell. The place of the carotenogenic response among the stress tolerance mechanisms is revisited, and possible implications of the new findings for biotechnological production of astaxanthin from microalgae are considered. The potential use of the carotenogenic microalgae as a source not only of value-added carotenoids, but also of biofuel precursors is discussed.

Preventive effect of dietary astaxanthin on UVA-induced skin photoaging in hairless mice

PubMed Central

Komatsu, Toshiyuki; Sasaki, Suguru; Manabe, Yuki; Hirata, Takashi

2017-01-01

Astaxanthin, a carotenoid found mainly in seafood, has potential clinical applications due to its antioxidant activity. In this study, we evaluated the effect of dietary astaxanthin derived from Haematococcus pluvialis on skin photoaging in UVA-irradiated hairless mice by assessing various parameters of photoaging. After chronic ultraviolet A (UVA) exposure, a significant increase in transepidermal water loss (TEWL) and wrinkle formation in the dorsal skin caused by UVA was observed, and dietary astaxanthin significantly suppressed these photoaging features. We found that the mRNA expression of lympho-epithelial Kazal-type-related inhibitor, steroid sulfatase, and aquaporin 3 in the epidermis was significantly increased by UVA irradiation for 70 days, and dietary astaxanthin significantly suppressed these increases in mRNA expression to be comparable to control levels. In the dermis, the mRNA expression of matrix metalloprotease 13 was increased by UVA irradiation and significantly suppressed by dietary astaxanthin. In addition, HPLC-PDA analysis confirmed that dietary astaxanthin reached not only the dermis but also the epidermis. Our results indicate that dietary astaxanthin accumulates in the skin and appears to prevent the effects of UVA irradiation on filaggrin metabolism and desquamation in the epidermis and the extracellular matrix in the dermis. PMID:28170435

Construction of transplastomic lettuce (Lactuca sativa) dominantly producing astaxanthin fatty acid esters and detailed chemical analysis of generated carotenoids.

PubMed

Harada, Hisashi; Maoka, Takashi; Osawa, Ayako; Hattan, Jun-Ichiro; Kanamoto, Hirosuke; Shindo, Kazutoshi; Otomatsu, Toshihiko; Misawa, Norihiko

2014-04-01

The plastid genome of lettuce (Lactuca sativa L.) cv. Berkeley was site-specifically modified with the addition of three transgenes, which encoded β,β-carotenoid 3,3'-hydroxylase (CrtZ) and β,β-carotenoid 4,4'-ketolase (4,4'-oxygenase; CrtW) from a marine bacterium Brevundimonas sp. strain SD212, and isopentenyl diphosphate isomerase from a marine bacterium Paracoccus sp. strain N81106. Constructed transplastomic lettuce plants were able to grow on soil at a growth rate similar to that of non-transformed lettuce cv. Berkeley and generate flowers and seeds. The germination ratio of the lettuce transformants (T0) (98.8%) was higher than that of non-transformed lettuce (93.1 %). The transplastomic lettuce (T1) leaves produced the astaxanthin fatty acid (myristate or palmitate) diester (49.2% of total carotenoids), astaxanthin monoester (18.2%), and the free forms of astaxanthin (10.0%) and the other ketocarotenoids (17.5%), which indicated that artificial ketocarotenoids corresponded to 94.9% of total carotenoids (230 μg/g fresh weight). Native carotenoids were there lactucaxanthin (3.8%) and lutein (1.3 %) only. This is the first report to structurally identify the astaxanthin esters biosynthesized in transgenic or transplastomic plants producing astaxanthin. The singlet oxygen-quenching activity of the total carotenoids extracted from the transplastomic leaves was similar to that of astaxanthin (mostly esterified) from the green algae Haematococcus pluvialis.

Xanthophyllomyces dendrorhous for the industrial production of astaxanthin.

PubMed

Rodríguez-Sáiz, Marta; de la Fuente, Juan Luis; Barredo, José Luis

2010-10-01

Astaxanthin is a red xanthophyll (oxygenated carotenoid) with large importance in the aquaculture, pharmaceutical, and food industries. The green alga Haematococcus pluvialis and the heterobasidiomycetous yeast Xanthophyllomyces dendrorhous are currently known as the main microorganisms useful for astaxanthin production at the industrial scale. The improvement of astaxanthin titer by microbial fermentation is a requirement to be competitive with the synthetic manufacture by chemical procedures, which at present is the major source in the market. In this review, we show how the isolation of new strains of X. dendrorhous from the environment, the selection of mutants by the classical methods of random mutation and screening, and the rational metabolic engineering, have provided improved strains with higher astaxanthin productivity. To reduce production costs and enhance competitiveness from an industrial point of view, low-cost raw materials from industrial and agricultural origin have been adopted to get the maximal astaxanthin productivity. Finally, fermentation parameters have been studied in depth, both at flask and fermenter scales, to get maximal astaxanthin titers of 4.7 mg/g dry cell matter (420 mg/l) when X. dendrorhous was fermented under continuous white light. The industrial scale-up of this biotechnological process will provide a cost-effective method, alternative to synthetic astaxanthin, for the commercial exploitation of the expensive astaxanthin (about $2,500 per kilogram of pure astaxanthin).

In Vitro Effects of Some Botanicals with Anti-Inflammatory and Antitoxic Activity

PubMed Central

Guidetti, Gianandrea; Giovazzino, Angela; Rubino, Valentina; Palatucci, Anna Teresa; Centenaro, Sara; Fraccaroli, Elena; Cortese, Laura; Bonomo, Maria Grazia; Ruggiero, Giuseppina; Canello, Sergio; Terrazzano, Giuseppe

2016-01-01

Several extrinsic factors, like drugs and chemicals, can foster autoimmunity. Tetracyclines, in particular oxytetracycline (OTC), appear to correlate with the emergence of immune-mediated diseases. Accumulation of OTC, the elective drug for gastrointestinal and respiratory infectious disease treatment in broiler chickens, was reported in chicken edible tissues and could represent a potential risk for pets and humans that could assume this antibiotic as residue in meat or in meat-derived byproducts. We investigated the in vitro anti-inflammatory properties of a pool of thirteen botanicals as a part of a nutraceutical diet, with proven immunomodulatory activity. In addition, we evaluated the effect of such botanicals in contrasting the in vitro proinflammatory toxicity of OTC. Our results showed a significant reduction in interferon- (INF-) γ production by human and canine lymphocytes in presence of botanicals (⁎ p < 0.05). Increased INF-γ production, dependent on 24-hour OTC-incubation of T lymphocytes, was significantly reduced by the coincubation with Haematococcus pluvialis, with Glycine max, and with the mix of all botanicals (⁎ p < 0.05). In conclusion, the use of these botanicals was shown to be able to contrast OTC-toxicity and could represent a new approach for the development of functional foods useful to enhance the standard pharmacological treatment in infections as well as in preventing or reducing the emergence of inflammatory diseases. PMID:27597982

Evolutionary origins, molecular cloning and expression of carotenoid hydroxylases in eukaryotic photosynthetic algae

PubMed Central

2013-01-01

Background Xanthophylls, oxygenated derivatives of carotenes, play critical roles in photosynthetic apparatus of cyanobacteria, algae, and higher plants. Although the xanthophylls biosynthetic pathway of algae is largely unknown, it is of particular interest because they have a very complicated evolutionary history. Carotenoid hydroxylase (CHY) is an important protein that plays essential roles in xanthophylls biosynthesis. With the availability of 18 sequenced algal genomes, we performed a comprehensive comparative analysis of chy genes and explored their distribution, structure, evolution, origins, and expression. Results Overall 60 putative chy genes were identified and classified into two major subfamilies (bch and cyp97) according to their domain structures. Genes in the bch subfamily were found in 10 green algae and 1 red alga, but absent in other algae. In the phylogenetic tree, bch genes of green algae and higher plants share a common ancestor and are of non-cyanobacterial origin, whereas that of red algae is of cyanobacteria. The homologs of cyp97a/c genes were widespread only in green algae, while cyp97b paralogs were seen in most of algae. Phylogenetic analysis on cyp97 genes supported the hypothesis that cyp97b is an ancient gene originated before the formation of extant algal groups. The cyp97a gene is more closely related to cyp97c in evolution than to cyp97b. The two cyp97 genes were isolated from the green alga Haematococcus pluvialis, and transcriptional expression profiles of chy genes were observed under high light stress of different wavelength. Conclusions Green algae received a β-xanthophylls biosynthetic pathway from host organisms. Although red algae inherited the pathway from cyanobacteria during primary endosymbiosis, it remains unclear in Chromalveolates. The α-xanthophylls biosynthetic pathway is a common feature in green algae and higher plants. The origination of cyp97a/c is most likely due to gene duplication before divergence of

Proceedings of Damping 89, Volume 2, West Palm Beach, FL, 8-10 February 1989

DTIC Science & Technology

1989-11-01

Vinyl Methyl Ether)-Polystyrene Blends and IPN’s J. J. Fay, Dr. C. J. Murphy, Dr. D. A. Thomas and Prof. L. H. Sperling BAA Time-Temperature...Structural Vibrations with Piezoelectric Materials and Passive Electrical Networks N. W. Hagood and Prof. A. von Flotow ICC Passive Electromagnetic

Conversion of stranded waste-stream carbon and nutrients into value-added products via metabolically coupled binary heterotroph-photoautotroph system

DOE Office of Scientific and Technical Information (OSTI.GOV)

Bohutskyi, Pavlo; Kucek, Leo A.; Hill, Eric

Metabolic flexibility and robustness of phototroph- heterotroph co-cultures provide a flexible binary engineering platform for a variety of biotechnological and environmental applications. Here, we metabolically coupled a heterotrophic bacterium Bacillus subtilis with astaxanthin producing alga Haematococcus pluvialis and successfully applied this binary co-culture for conversion of the starch-rich waste stream into valuable astaxanthin-rich biomass. Importantly, the implemented system required less mass transfer of CO2 and O2 due to in-situ exchange between heterotroph and phototroph, which can contribute to reduction in energy consumption for wastewater treatment. In addition, the maximum reduction in chemical oxygen demand, total nitrogen and phosphorus reached 65%,more » 55% and 30%, respectively. The preliminary economic analysis indicated that realization of produced biomass with 0.8% astaxanthin content may generate annual revenues of $3.2M (baseline scenario) from treatment of wastewater (1,090 m3/day) from a potato processing plant. Moreover, the revenues may be increased up to $18.2M for optimized scenario with astaxanthin content in algae of 2%. This work demonstrates a successful proof-of-principle for conversion of waste carbon and nutrients into targeted value-added products through metabolic connection of heterotrophic and phototrophic organisms. Utilization of heterotrophic-algal binary cultures opens new perspectives for designing highly-efficient production processes for feedstock biomass production as well as allows utilization of variety of organic agricultural, chemical, or municipal wastes.« less

Determination of fucoxanthin isomers in microalgae (Isochrysis sp.) by high-performance liquid chromatography coupled with diode-array detector multistage mass spectrometry coupled with positive electrospray ionization.

PubMed

Crupi, Pasquale; Toci, Aline Theodore; Mangini, Silvio; Wrubl, Federico; Rodolfi, Liliana; Tredici, Mario R; Coletta, Antonio; Antonacci, Donato

2013-05-15

Due to their health benefits, there is growing interest in the production and use of carotenoids from natural sources, e.g. microalgae. To date, only Haematococcus pluvialis and Dunaliella, that accumulate, respectively, astaxanthin and β-carotene in large quantities, are grown commercially. However, interest is also being focused on other xanthophylls, such as (all-E)-fucoxanthin characterized by anti-obesity and anti-carcinogenic effects. In this regard, rigorous chemical and analytical techniques following preparative isolation of components are needed to unequivocally identify individual carotenoids in microalgae. The carotenoid profile of Isochrysis sp. biomass, produced in closed photobioreactors, was determined by reversed-phase C30 (RP-30) high-performance liquid chromatography coupled with diode-array detector mass spectrometry using positive electrospray ionization (HPLC/DAD-MS/ESI(+) ) analysis. Additionally, multistage mass spectrometry (MS(n) ) analyses, together with fine structures of the UV-vis spectra, were used to differentiate structural and geometrical isomers. This technique allowed the simultaneous determination of geometrical, isomers of fucoxanthin (all-E-fucoxanthin, 13Z-, 13'Z- and 9'Z-fucoxanthin), diatoxanthin and 5,8-epoxydiadinoxanthin diasteroisomers (R/S). The analyzed extracts contained fucoxanthin isomers as the major carotenoids and, in particular, (all-E)-fucoxanthin was the main geometrical isomer (~85%) found at a concentration of 17 mg/g of the lyophilized biomass. Considering the high content of fucoxanthin in Isochrysis sp. biomass, the microalga could be proposed as a source of this compound for nutraceutical and pharmaceutical applications. Copyright © 2013 John Wiley & Sons, Ltd.

The Effect of Astaxanthin-Rich Microalgae "Haematococcus pluvialis" and Wholemeal Flours Incorporation in Improving the Physical and Functional Properties of Cookies.

PubMed

Hossain, A K M Mofasser; Brennan, Margaret A; Mason, Susan L; Guo, Xinbo; Zeng, Xin An; Brennan, Charles S

2017-07-26

Marine-based food supplements can improve human nutrition. In an effort to modulate glycaemic response and enhance nutritional aspects, marine-derived algal food rich in astaxanthin was used in the formulation of a model food (wholemeal cookie). Astaxanthin substitution of cookies made from three flours (wheat, barley and oat) demonstrated a significant reduction in the rate of glucose released during in vitro digestion together with an increase in the total phenolic content (TPC) and antioxidant capacity of the food. The significantly ( p < 0.005) lower free glucose release was observed from cookies with 15% astaxanthin, followed by 10% and then 5% astaxanthin in comparison with control cookies of each flour. Total phenolic content, DPPH radical scavenging and Oxygen Radical Absorbance Capacity (ORAC) value also notably increased with increase in astaxanthin content. The results evidence the potential use of microalgae to enhance the bioactive compounds and lower the glycaemic response of wholemeal flour cookie.

Mussel processing wastewater: a low-cost substrate for the production of astaxanthin by Xanthophyllomyces dendrorhous.

PubMed

Amado, Isabel Rodríguez; Vázquez, José Antonio

2015-11-09

The use of astaxanthin in different industries such as the chemical, pharmaceutical, food, animal feed and cosmetic has been receiving increasing attention in recent years. Natural supplies of the pigment include crustacean by-products, algal, and microbial cultivation, being the yeast Xanthophyllomyces dendrorhous together with the alga Haematococcus pluvialis the most promising microorganisms for this bioproduction. Different vegetable by-products of the food industry have been explored so far as low-cost substrates for the production of astaxanthin by X. dendrorhous. This study focuses for the first time on the use of a low-cost formulated medium from a marine by-product, mussel-processing wastewater, for the production of astaxanthin by the yeast X. dendrorhous. The yeast was able to grow in non-saccharified mussel broth, revealing the ability of the microorganism to hydrolyze glycogen. However, partial glycogen saccharification with α-amylase was needed for astaxanthin biosynthesis, obtaining maximal productions of 22.5-26.0 mg/L towards the end of the culture and coinciding with yeast highest amylolytic activity. Cultivations in totally-saccharified media revealed an increase in maximal cell concentrations and a decrease in maximal growth rates and astaxanthin production with increasing glucose initial concentration. Astaxanthin production was higher in partially-saccharified mussel-processing waste than in synthetic medium (yeast peptone dextrose) containing glucose as carbon source (13 mg/L), suggesting this by-product is a promising nutritive medium for astaxanthin production. The use of this effluent also contributes towards the recycling and depuration of this highly pollutant effluent.

Functional characterization of various algal carotenoid ketolases reveals that ketolating zeaxanthin efficiently is essential for high production of astaxanthin in transgenic Arabidopsis

PubMed Central

Zhong, Yu-Juan; Huang, Jun-Chao; Liu, Jin; Li, Yin; Jiang, Yue; Xu, Zeng-Fu; Sandmann, Gerhard; Chen, Feng

2011-01-01

Extending the carotenoid pathway to astaxanthin in plants is of scientific and industrial interest. However, expression of a microbial β-carotene ketolase (BKT) that catalyses the formation of ketocarotenoids in transgenic plants typically results in low levels of astaxanthin. The low efficiency of BKTs in ketolating zeaxanthin to astaxanthin is proposed to be the major limitation for astaxanthin accumulation in engineered plants. To verify this hypothesis, several algal BKTs were functionally characterized using an Escherichia coli system and three BKTs were identified, with high (up to 85%), moderate (∼38%), and low (∼1%) conversion rate from zeaxanthin to astaxanthin from Chlamydomonas reinhardtii (CrBKT), Chlorella zofingiensis (CzBKT), and Haematococcus pluvialis (HpBKT3), respectively. Transgenic Arabidopsis thaliana expressing the CrBKT developed orange leaves which accumulated astaxanthin up to 2 mg g−1 dry weight with a 1.8-fold increase in total carotenoids. In contrast, the expression of CzBKT resulted in much lower astaxanthin content (0.24 mg g−1 dry weight), whereas HpBKT3 was unable to mediate synthesis of astaxanthin in A. thaliana. The none-native astaxanthin was found mostly in a free form integrated into the light-harvesting complexes of photosystem II in young leaves but in esterified forms in senescent leaves. The alteration of carotenoids did not affect chlorophyll content, plant growth, or development significantly. The astaxanthin-producing plants were more tolerant to high light as shown by reduced lipid peroxidation. This study advances a decisive step towards the utilization of plants for the production of high-value astaxanthin. PMID:21398427

Proceedings of Damping 󈨟, Held in San Diego, California on 13 - 15 February 1991. Volume 2

DTIC Science & Technology

1991-08-01

Efficiency CBC* Dr. L. H. Sperling , J. J. Fay, and Dr. D. A. Thomas The Thermorheologically Complex Material CBD* Lt. Col. Ronald L. Bagley SESSION CC...Andrew S. Bicos. and J. S. Fechter The Need for Passive Damping in Feedback Controlled Flexible Struc- GBB tures Dr. Andreas von Flotow and D. W. Vos...Vibration Research, Southampton, England (R.C. Drew) Lehigh University, Bethlehem, Pennsylvania (L. Sperling ) MrS Corporation, Minneapolis, Minnesota

Ketocarotenoid Production in Soybean Seeds through Metabolic Engineering

PubMed Central

Pierce, Emily C.; LaFayette, Peter R.; Ortega, María A.; Joyce, Blake L.; Kopsell, Dean A.; Parrott, Wayne A.

2015-01-01

The pink or red ketocarotenoids, canthaxanthin and astaxanthin, are used as feed additives in the poultry and aquaculture industries as a source of egg yolk and flesh pigmentation, as farmed animals do not have access to the carotenoid sources of their wild counterparts. Because soybean is already an important component in animal feed, production of these carotenoids in soybean could be a cost-effective means of delivery. In order to characterize the ability of soybean seed to produce carotenoids, soybean cv. Jack was transformed with the crtB gene from Pantoea ananatis, which codes for phytoene synthase, an enzyme which catalyzes the first committed step in the carotenoid pathway. The crtB gene was engineered together in combinations with ketolase genes (crtW from Brevundimonas sp. strain SD212 and bkt1 from Haematococcus pluvialis) to produce ketocarotenoids; all genes were placed under the control of seed-specific promoters. HPLC results showed that canthaxanthin is present in the transgenic seeds at levels up to 52 μg/g dry weight. Transgenic seeds also accumulated other compounds in the carotenoid pathway, such as astaxanthin, lutein, β-carotene, phytoene, α-carotene, lycopene, and β-cryptoxanthin, whereas lutein was the only one of these detected in non-transgenic seeds. The accumulation of astaxanthin, which requires a β-carotene hydroxylase in addition to a β-carotene ketolase, in the transgenic seeds suggests that an endogenous soybean enzyme is able to work in combination with the ketolase transgene. Soybean seeds that accumulate ketocarotenoids could potentially be used in animal feed to reduce or eliminate the need for the costly addition of these compounds. PMID:26376481

Proceedings of Damping󈨝 Held in West Palm Beach, Florida on 8-10 February 1989. Volume 2. Pages FAA-1 through IBD-15)

DTIC Science & Technology

1989-11-01

Behavior of Poly (Vinyl Methyl Ether)-Polystyrene Blends and IEN’s J. J. Fay, Dr. C. J. Murphy, Dr. D. A. Thomas and Prof. L. H. Sperling BAA Time...ICB Damping of Structural Vibrations with Piezoelectric Materials and Passive Electrical Networks N. W. Hagood and Prof. A. von Flotow ICC Passive...Fri9 = - d-T- (10) and Fti = e L "d + e I (11) The flux terms Rri and Rti are given by ait* Von dS+f t* (V’n) dSRri = - Saf4csae ___ c 1L 1 (V.n

Creating a Collection of Microalgae for use in Biofuels Research

DTIC Science & Technology

2008-06-25

Chlorella , lower left – Scenedesmus, lower center – Fragillaria, lower right – Chlamydomonad. Figure 7: Shown is a phylogram based on the nuclear...variety of desmids including but not limited to Scenedesmus, Tetradesmus, Chlamydomonas, Chlorella , Chlorococcum, Haematococcus, and Kirchneriella

Inorganic and organic contaminants in Alaskan shorebird eggs.

PubMed

Saalfeld, David T; Matz, Angela C; McCaffery, Brian J; Johnson, Oscar W; Bruner, Phil; Lanctot, Richard B

2016-05-01

Many shorebird populations throughout North America are thought to be declining, with potential causes attributed to habitat loss and fragmentation, reduced prey availability, increased predation, human disturbance, and increased exposure to environmental pollutants. Shorebirds may be particularly vulnerable to contaminant exposure throughout their life cycle, as they forage primarily on invertebrates in wetlands, where many contaminants accumulate disproportionately in the sediments. Therefore, it is important to document and monitor shorebird populations thought to be at risk and assess the role that environmental contaminants may have on population declines. To investigate potential threats and provide baseline data on shorebird contaminant levels in Alaskan shorebirds, contaminant concentrations were evaluated in shorebird eggs from 16 species residing in seven geographic distinct regions of Alaska. Similar to previous studies, low levels of most inorganic and organic contaminants were found, although concentrations of several inorganic and organic contaminants were higher than those of previous studies. For example, elevated strontium levels were observed in several species, especially black oystercatcher (Haematopus bachmani) sampled in Prince William Sound, Alaska. Additionally, contaminant concentrations varied among species, with significantly higher concentrations of inorganic contaminants found in eggs of pectoral sandpiper (Calidris melanotos), semipalmated sandpiper (Calidris pusilla), black oystercatcher, and bar-tailed godwit (Limosa lapponica). Similarly, significantly higher concentrations of some organic contaminants were found in the eggs of American golden plover (Pluvialis dominica), black-bellied plover (Pluvialis squatarola), pacific golden plover (Pluvialis fulva), bar-tailed godwit, and semipalmated sandpiper. Despite these elevated levels, current concentrations of contaminants in shorebird eggs suggest that breeding environments are

Response of Tabanidae (Diptera) to different natural attractants.

PubMed

Krcmar, Stjepan; Mikuska, Alma; Merdić, Enrih

2006-12-01

The response of female tabanids to natural attractants was studied in the Monjoros Forest along the Nature Park Kopacki rit in eastern Croatia. Tabanids were caught in canopy traps baited with either aged cow, horse, sheep, or pig urine and also in unbaited traps. Tabanids were collected in a significantly higher numbers in traps baited with natural attractants compared to unbaited traps. The number of females of Tabanus bromius, Tabanus maculicornis, Tabanus tergestinus, and Hybomitra bimaculata collected from canopy traps baited with cow urine and traps baited with other natural attractants differed significantly. Females of Haematopota pluvialis were also collected more frequently in canopy traps baited with aged cow urine than in those with aged horse urine, but this difference was not significant. However, the number of females of Haematopota pluvialis collected from canopy traps baited with other natural attractants (sheep and pig urine) differed significantly when compared with aged cow urine baited traps. Canopy traps baited with aged cow urine collected significantly more Tabanus sudeticus than did traps baited with aged pig urine. Finally, the aged cow urine baited canopy traps collected 51 times more tabanids than unbaited traps, while aged horse, aged sheep, and aged pig urine baited traps collected 36, 30, and 22 times as many tabanids, respectively, than unbaited traps.

A new species of Leptolalax (Amphibia, Anura, Megophryidae) from Peninsular Malaysia.

PubMed

Matsui, Masafumi; Belabut, Daicus M; Ahmad, Norhayati; Yong, Hoi-Sen

2009-03-01

A new megophryid species, Leptolalax kecil, is described from the Cameron Highlands of Peninsular Malaysia on the basis of acoustic and morphological characteristics. It has an advertisement call dissimilar to that of other, congeneric species, and is the smallest of the known Leptolalax. It is similar to L. pluvialis from Vietnam in small body size, but differs from it in ventral color, and in the size and color of the pectoral gland. The distributional pattern of the new species Is discussed.

[Simultaneous determination of carotenoids and chlorophylls in algae by high performance liquid chromatography].

PubMed

Yuan, J; Zhang, Y; Shi, X; Gong, X; Chen, F

1997-03-01

An isocratic reversed-phase HPLC method for the simultaneous determination of carotenoids and chlorophylls in algae is presented. Both carotenoids and chlorophylls were separated by using a C18 column and a mobile phase consisting of dichloromethane/acetonitrile/methanol/water (22.5:9.5:67.5:0.5). The flow rate was 1.0mL/min. These pigments were detected by a UV-VIS absorbance detector set at 450nm. The average recoveries of these pigments by standard addition method were 99.1% for lutein, 98.5% for alpha-carotene, 99.4% for beta-carotene, 100.6% for chlorophyll a and 99.9% for chlorophyll b. The relative standard deviations of these pigments were 2.4% for lutein, 5.6% for alpha-carotene, 6.0% for beta-carotene, 4.1% for chlorophyll a and 4.0% for chlorophyll b. The calibration curves for these pigments gave good linearity over a wide range of 0-50mg/L (r = 0.9992-0.9999). The retention times were 3.0 min for lutein, 4.7 min for chlorophyll b, 6.3 min for chlorophyll a, 14.1 min for alpha-carotene, and 14.7 min for beta-carotene. Using this method, carotenoids and chlorophylls were determined in Spirulina platensis, Chlorella vulgaris and Haematococcus lacustris. Chlorella vulgaris and Haematococcus lacustris samples were ground in a B. Braun homogenizer. Spirulina platensis sample was smashed by supersonic wave. Acetone was used as extractant.

A new species of Pristimantis (Amphibia, Anura, Craugastoridae) from the foothills of the Andes in Manu National Park, southeastern Peru

PubMed Central

Shepack, Alexander; von May, Rudolf; Ttito, Alex; Catenazzi, Alessandro

2016-01-01

Abstract We describe a new species of Pristimantis from the humid sub-montane forest of the Región Cusco in Peru. Pristimantis pluvialis sp. n. was collected in the Kosñipata and Entoro valleys at elevations from 740 to 1110 m a.s.l., near the borders of Manu National Park and within the Huachiperi Haramba Queros Conservation Concession. The new species can be distinguished from other members of the genus Pristimantis by its rostral tubercle, smooth dorsal skin, and by its advertisement call. Pristimantis lacrimosus and Pristimantis waoranii superficially most resemble the new species, but Pristimantis pluvialis sp. n. differs from both species by having a rostral tubercle (absent in Pristimantis waoranii and variable in Pristimantis lacrimosus) and larger size, from Pristimantis lacrimosus by its call emitted at a lower frequency, and from Pristimantis waoranii for its dorsal coloration with dark markings. Two other species have partially overlapping distributions and resemble the new species, Pristimantis mendax and Pristimantis olivaceus, but they produce advertisement calls with much higher dominant frequencies than the advertisement call of the new species. Furthermore, Pristimantis mendax differs from the new species by lacking a rostral tubercle and by having a sigmoid inner tarsal fold, whereas Pristimantis olivaceus differs by being smaller and by having dorsal skin shagreen with scattered tubercles. The new species has snout-vent length of 21.8–26.9 mm in males (n = 12) and 28.8 mm in a single female. PMID:27408563

A new species of Pristimantis (Amphibia, Anura, Craugastoridae) from the foothills of the Andes in Manu National Park, southeastern Peru.

PubMed

Shepack, Alexander; von May, Rudolf; Ttito, Alex; Catenazzi, Alessandro

2016-01-01

We describe a new species of Pristimantis from the humid sub-montane forest of the Región Cusco in Peru. Pristimantis pluvialis sp. n. was collected in the Kosñipata and Entoro valleys at elevations from 740 to 1110 m a.s.l., near the borders of Manu National Park and within the Huachiperi Haramba Queros Conservation Concession. The new species can be distinguished from other members of the genus Pristimantis by its rostral tubercle, smooth dorsal skin, and by its advertisement call. Pristimantis lacrimosus and Pristimantis waoranii superficially most resemble the new species, but Pristimantis pluvialis sp. n. differs from both species by having a rostral tubercle (absent in Pristimantis waoranii and variable in Pristimantis lacrimosus) and larger size, from Pristimantis lacrimosus by its call emitted at a lower frequency, and from Pristimantis waoranii for its dorsal coloration with dark markings. Two other species have partially overlapping distributions and resemble the new species, Pristimantis mendax and Pristimantis olivaceus, but they produce advertisement calls with much higher dominant frequencies than the advertisement call of the new species. Furthermore, Pristimantis mendax differs from the new species by lacking a rostral tubercle and by having a sigmoid inner tarsal fold, whereas Pristimantis olivaceus differs by being smaller and by having dorsal skin shagreen with scattered tubercles. The new species has snout-vent length of 21.8-26.9 mm in males (n = 12) and 28.8 mm in a single female.

Seasonal abundance of horse flies (Diptera: Tabanidae) from two locations in eastern Croatia.

PubMed

Krcmar, Stjepan

2005-12-01

A total of 10,539 tabanid horse flies from 22 species and five genera was collected in the Tikves forest within the Kopacki rit Nature Park in eastern Croatia. Seasonal abundance was analyzed for the six most abundant species. Tabanus maculicornis, Tabanus tergestinus, and Haematopota pluvialis reached their highest peak abundance in the fourth week of June. Atylotus loewianus and Tabanus bromius reached their highest peak of abundance in the first week of August, whereas Tabanus sudeticus reached its maximum abundance in the third week of July. Horse flies also were collected once a week on the pasture at Petrijevci from mid-May to mid-September during 1993. Paired collections were made from a Malaise trap and from a horse by using a sweep net. A total of 2,867 tabanids belonging to 26 species was collected. The number of tabanids collected on horses was much higher than the total captured with Malaise traps. On their natural host (horse), 2.6 times more tabanids were collected than in the traps. Seasonal abundance was analyzed only for the eight most abundant species. Chrysops paralellogrammus, Tabanus autumnalis, Tabanus bromius, Tabanus tergestinus, Haematopota pluvialis, and Haematopota subcylindrica all reached their highest peak of abundance in the second week of July, whereas Tabanus maculicornis reached the maximal peak of abundance in the third week of June. Seasonal meteorological variability that occurs periodically from one year to another has a significant influence on the maximal peaks of tabanid abundance.

Predicting breeding shorebird distributions on the Arctic Coastal Plain of Alaska

USGS Publications Warehouse

Saalfeld, Sarah T.; Lanctot, Richard B.; Brown, Stephen C.; Saalfeld, David T.; Johnson, James A.; Andres, Brad A.; Bart, Jonathan R.

2013-01-01

The Arctic Coastal Plain (ACP) of Alaska is an important region for millions of migrating and nesting shorebirds. However, this region is threatened by climate change and increased human development (e.g., oil and gas production) that have the potential to greatly impact shorebird populations and breeding habitat in the near future. Because historic data on shorebird distributions in the ACP are very coarse and incomplete, we sought to develop detailed, contemporary distribution maps so that the potential impacts of climate-mediated changes and development could be ascertained. To do this, we developed and mapped habitat suitability indices for eight species of shorebirds (Black-bellied Plover [Pluvialis squatarola], American Golden-Plover [Pluvialis dominica], Semipalmated Sandpiper [Calidris pusilla], Pectoral Sandpiper [Calidris melanotos], Dunlin [Calidris alpina], Long-billed Dowitcher [Limnodromus scolopaceus], Red-necked Phalarope [Phalaropus lobatus], and Red Phalarope [Phalaropus fulicarius]) that commonly breed within the ACP of Alaska. These habitat suitability models were based on 767 plots surveyed during nine years between 1998 and 2008 (surveys were not conducted in 2003 and 2005), using single-visit rapid area searches during territory establishment and incubation (8 June, 1 July). Species specific habitat suitability indices were developed and mapped using presence-only modeling techniques (partitioned Mahalanobis distance) and landscape environmental variables. For most species, habitat suitability was greater at lower elevations (i.e., near the coast and river deltas) and lower within upland habitats. Accuracy of models was high for all species, ranging from 65 -98%. Our models predicted that the largest fraction of suitable habitat for the majority of species occurred within the National Petroleum Reserve-Alaska, with highly suitable habitat also occurring within coastal areas of the Arctic National Wildlife Refuge west to Prudhoe Bay.

The complete mitochondrial genome of Gryllotalpa unispina Saussure, 1874 (Orthoptera: Gryllotalpoidea: Gryllotalpidae).

PubMed

Zhang, Yulong; Shao, Dandan; Cai, Miao; Yin, Hong; Zhang, Daochuan

2016-01-01

The complete mitochondrial genome of Gryllotalpa unispina was 15,513 bp in length and contained 70.9% AT. All G. unispina protein-coding sequences except for the nad2 started with a typical ATN codon. The usual termination codons (TAA) and incomplete stop codons (T) were found from 13 protein-coding genes. All tRNA genes were folded into the typical cloverleaf secondary structure, except trnS(AGN) lacking the dihydrouridine arm. The sizes of the large and small ribosomal RNA genes were 1245 and 725 bp, respectively. The A + T-rich region was 917 bp in length with 76.8%. The orientation and gene order of the G. unispina mitogenome were identical to the G. orientalis and G. pluvialis, there was no phenomenon of "DK rearrangement" which has been widely reported in Caelifera.

Cell disruption and lipid extraction for microalgal biorefineries: A review.

PubMed

Lee, Soo Youn; Cho, Jun Muk; Chang, Yong Keun; Oh, You-Kwan

2017-11-01

The microalgae-based biorefinement process has attracted much attention from academic and industrial researchers attracted to its biofuel, food and nutraceutical applications. In this paper, recent developments in cell-disruption and lipid-extraction methods, focusing on four biotechnologically important microalgal species (namely, Chlamydomonas, Haematococcus, Chlorella, and Nannochloropsis spp.), are reviewed. The structural diversity and rigidity of microalgal cell walls complicate the development of efficient downstream processing methods for cell-disruption and subsequent recovery of intracellular lipid and pigment components. Various mechanical, chemical and biological cell-disruption methods are discussed in detail and compared based on microalgal species and status (wet/dried), scale, energy consumption, efficiency, solvent extraction, and synergistic combinations. The challenges and prospects of the downstream processes for the future development of eco-friendly and economical microalgal biorefineries also are outlined herein. Copyright © 2017 Elsevier Ltd. All rights reserved.

[Microalgae as the source of natural products].

PubMed

Vasas, Gábor

2018-05-01

More than 90% of herbal products and herbal medicines have been derived from higher plants recently, but due to independent circumstances, several photosynthetic microalgal species are in focus in this point of view. In the last 50 years, many carbohydrate-, peptide-, terpenoid-, alkaloid- and phenol-type components were described from algae because of the developing structural determination and analytical methods, algae mass production and also artificial algae technologies. At the same time, based partly on traditional causes and partly on the clinical and preclinical data of today, some dried products of algae are directly used as food supplements. Hereinafter, the historical background, economic significance and metabolic background of the mostly used microalgal species will be reviewed. The diverse metabolite production of these organisms will be demonstrated by some molecules with special bioactivity. Several preclinical and clinical studies will be described relating to the microalgal species Spirulina sp., Chlorella sp., Haematococcus sp. and Dunaliella sp. Orv Hetil. 2018; 159(18): 703-708.

Maximizing carotenoid extraction from microalgae used as food additives and determined by liquid chromatography (HPLC).

PubMed

Cerón-García, M C; González-López, C V; Camacho-Rodríguez, J; López-Rosales, L; García-Camacho, F; Molina-Grima, E

2018-08-15

Microalgae are an interesting source of natural pigments that have valuable applications. However, further research is necessary to develop processes that allow us to achieve high levels of carotenoid recovery while avoiding degradation. This work presents a comprehensive study on the recovery of carotenoids from several microalgae genera, optimizing carotenoid extraction using alkaline saponification at various temperatures and KOH concentrations. Results show that I. galbana requires a temperature of 60 °C and <10% KOH, N. gaditana and K. veneficum require 60 °C and no saponification, P. reticulatum requires 40 °C and 10% KOH, T. suecica and H. pluvialis require 25 °C and 40% KOH while C. sp. and S. almeriensis require 80 °C and 40% KOH. The influence of the solvent on carotenoid recovery was also studied. In general terms, an ethanol:hexane:water (77:17:6 v/v/v) mixture results in good yields. Copyright © 2018 Elsevier Ltd. All rights reserved.

Extraction of astaxanthin from microalgae: process design and economic feasibility study

NASA Astrophysics Data System (ADS)

Zgheib, Nancy; Saade, Roxana; Khallouf, Rindala; Takache, Hosni

2018-03-01

In this work, the process design and the economic feasibility of natural astaxanthin extraction fromHaematococcus pluvialisspecies have been reported. Complete process drawing of the process was first performed, and then the process was designed including five main steps being the harvesting process, the cell disruption, the spray drying, the supercritical CO2extraction and the anaerobic digestion. The major components of the facility would include sedimentation tanks, a disk stack centrifuge, a bed miller, a spray dryer, a multistage compressor, an extractor, a pasteurizer and a digester. All units have been sized assuming a 10 kg/h of dried biomass as a feedstock to produce nearly 2592 kg of astaxanthin per year. The investment payback time and the return on investment were all estimated for different market prices of astaxanthin. Based on the results the production process was found to become economically feasible for a market price higher than 1500/Kg. Also, a payback period of 1 year and an ROI equal to 113% was estimated for an astaxanthin market price equal to 6000/Kg.

Evaluation of algal phytodegradation of petroleum naphthenic acids.

PubMed

Headley, John V; Du, Jing Long; Peru, Kerry M; Gurprasad, Narine; McMartin, Dena W

2008-02-15

The algal phytodegradation of a model naphthenic acid (4-methylcyclohexaneacetic acid) and an oilsands mixture of naphthenic acids (NAs) were evaluated in support of studies to remediate recalcitrant NAs in soils and water. The algae investigated included blue-green algae (Oscillatoria sp.; Aphanizomenon sp.; Anbaena sp.) green algae (Selenastrum sp.; Nannochloris sp.; Ankistrodesmus sp.; Scenedesmus sp.; Haematococcus sp.; Chlorella sp.) and diatoms (Naviculla (1), Naviculla (2) and Nitzschia sp.). Both the cis- and trans-isomers of the model NA were completely uptaken and presumed phytodegraded by the diatom algae Naviculla (2) sp. at a concentration of approximately 5.5 mg/L within a period of 14 days. However, there was no evidence for the phytodegradation of the petroleum oilsands naphthenic acids mixtures, except for possibly experiments utilizing the green algae, Selenastrum sp. The differences in the phytodegradation of the model NA by the diatoms appears to be linked to differences in transport mechanisms by the algae along with differences in the concentration and structure of the respective naphthenic acids.

Identification of a Novel Esterase from Marine Environmental Genomic DNA Libraries and Its Application in Production of Free All- trans-Astaxanthin.

PubMed

Lu, Ping; Gao, Xinwei; Dong, Hao; Liu, Zhen; Secundo, Francesco; Xue, Changhu; Mao, Xiangzhao

2018-03-21

Astaxanthin is a pigment with various functions. Free astaxanthin is obtained mainly through saponification methods, which could result in many byproducts. Enzymatic methods using lipases have been used in a few cases, while there are no reports on the use of esterases for the production of free astaxanthin. Herein we present the screening and identification of a novel esterase (Est3-14) from a marine mud metagenomic library. Est3-14 is pH-sensitive and keeps good stability in alkaline buffers (residual activity 94%, pH 8.0, 4 °C, and 36 h). Meanwhile, Est3-14 keeps a good stability in the medium temperature condition (residual activity 56.7%, pH 8.0, 40 °C, and 84 h). Est3-14 displayed high hydrolysis activity to prepare free all- trans-astaxanthin in biphasic systems. Furthermore, under optimal conditions (0.5 mL ethanol, 6 mL 0.1 M Tris-HCl buffer, pH 8.0, 0.5% (w/v) H. pluvialis oil, 40 °C), the hydrolytic conversion ratio was 99.3% after 36 h.

Brood amalgamation in the Bristle-thighed Curlew Numenius tahitiensis: process and function

USGS Publications Warehouse

Lanctot, Richard B.; Gill, Robert E.; Tibbitts, T.L.; Handel, Colleen M.

1995-01-01

Alloparental care in birds generally involves nonbreeding adults that help at nests or breeding adults that help raise young in communal nests. A less often reported form involves the amalgamation of broods, where one or more adults care for young that are not their own. We observed this phenomenon among Bristle-thighed Curlew Numenius tahitiensis broods in western Alaska during 1990–1992. Amalgamation of broods generally involved the formation of temporary and extended associations. Temporary associations were formed by the incidental convergence of broods soon after they left their nests. During this period, parents defended distinct brood-rearing areas, were antagonistic to conspecifics and remained together for less than 3 days. Extended associations formed when chicks were 1–2 weeks old. Here, parents and their broods occupied distinct, but adjacent, brood-rearing areas and moved around as a unit. Whether a brood participated in either temporary or extended associations or remained solitary appeared to depend on brood density in the immediate area and on hatching date. When chicks were 3–4 weeks old, aggregations of up to ten broods formed wherein young mixed and parents defended a common brood-rearing area. All broods (n = 48) that survived to fledging joined such aggregations. Alloparental care involved only antipredator defence and was not associated with activities such as feeding and brooding. Most female parents abandoned their broods shortly after the young could fly and when aggregations were forming. The female parent of a pair always deserted its young before or on the same day as the male parent and, in every aggregation, one or two males continued to tend young for about 5 days longer than other male parents. In most cases, adults deserted the young 2–6 days before the young departed the area when about 38 days old. Bristle-thighed Curlews also formed temporary associations with American and Pacific Golden Plover Pluvialis dominica and

Metabolic engineering of Dunaliella salina for production of ketocarotenoids.

PubMed

Anila, N; Simon, Daris P; Chandrashekar, Arun; Ravishankar, G A; Sarada, R

2016-03-01

Dunaliella is a commercially important marine alga producing high amount of β-carotene. The use of Dunaliella as a potential transgenic system for the production of recombinant proteins has been recently recognized. The present study reports for the first time the metabolic engineering of carotenoid biosynthesis in Dunaliella salina for ketocarotenoid production. The pathway modification included the introduction of a bkt gene from H. pluvialis encoding β-carotene ketolase (4,4'β-oxygenase) along with chloroplast targeting for the production of ketocarotenoids. The bkt under the control of Dunaliella Rubisco smaller subunit promoter along with its transit peptide sequence was introduced into the alga through standardized Agrobacterium-mediated transformation procedure. The selected transformants were confirmed using GFP and GUS expression, PCR and southern blot analysis. A notable upregulation of the endogenous hydroxylase level of transformants was observed where the BKT expression was higher in nutrient-limiting conditions. Carotenoid analysis of the transformants through HPLC and MS analysis showed the presence of astaxanthin and canthaxanthin with maximum content of 3.5 and 1.9 µg/g DW, respectively. The present study reports the feasibility of using D. salina for the production of ketocarotenoids including astaxanthin.

Dietary Carotenoids Regulate Astaxanthin Content of Copepods and Modulate Their Susceptibility to UV Light and Copper Toxicity

PubMed Central

Caramujo, Maria-José; de Carvalho, Carla C. C. R.; Silva, Soraya J.; Carman, Kevin R.

2012-01-01

High irradiation and the presence of xenobiotics favor the formation of reactive oxygen species in marine environments. Organisms have developed antioxidant defenses, including the accumulation of carotenoids that must be obtained from the diet. Astaxanthin is the main carotenoid in marine crustaceans where, among other functions, it scavenges free radicals thus protecting cell compounds against oxidation. Four diets with different carotenoid composition were used to culture the meiobenthic copepod Amphiascoides atopus to assess how its astaxanthin content modulates the response to prooxidant stressors. A. atopus had the highest astaxanthin content when the carotenoid was supplied as astaxanthin esters (i.e., Haematococcus meal). Exposure to short wavelength UV light elicited a 77% to 92% decrease of the astaxanthin content of the copepod depending on the culture diet. The LC50 values of A. atopus exposed to copper were directly related to the initial astaxanthin content. The accumulation of carotenoids may ascribe competitive advantages to certain species in areas subjected to pollution events by attenuating the detrimental effects of metals on survival, and possibly development and fecundity. Conversely, the loss of certain dietary items rich in carotenoids may be responsible for the amplification of the effects of metal exposure in consumers. PMID:22822352

Responses of Tabanidae (Diptera) to canopy traps baited with 4-methylphenol, 3-isopropylphenol, and naphthalene.

PubMed

Krcmar, Stjepan

2007-12-01

The attraction of female tabanids to unbaited and single-baited canopy traps using 4-methylphenol, 3-isopropylphenol, and naphthalene was studied in three forest localities in eastern Croatia. Tabanids were collected in a significantly higher number in traps baited with these chemicals compared to unbaited control traps. The number of females of Tabanus bromius, Tabanus sudeticus, Tabanus tergestinus, Hybomitra ciureai, Haematopota pluvialis, and Tabanus maculicornis collected from 4-methylphenol baited canopy traps and traps baited with other attractants differed significantly. A total of 89.0% of tabanids collected belonged to these six species. The response of the other species to used chemicals was not analyzed because of small sample sizes. Moreover, the results with 3-isopropylphenol and naphthalene are very similar and not significant for some tabanids. Tabanus bromius was the most abundant species with 48.4% in the sample collected by canopy traps. Finally, the 4-methylphenol baited canopy traps collected 16 times more tabanids than unbaited traps, while 3-isopropylphenol and naphthalene baited traps collected 3.5 and 2 times as many tabanids, respectively, than unbaited traps. Also, 4-methylphenol appeared to be a very effective attractant for Lucilia caesar (Calliphoridae), Sarcophaga carnaria (Sarcophagidae), and Musca domestica (Muscidae).

Birds of the Kilbuck and Ahklun mountain region, Alaska

USGS Publications Warehouse

Petersen, Margaret R.; Weir, Douglas N.; Dick, Matthew H.

1991-01-01

Between 1952 and 1988, we studied the abundance, distribution, occurrence, and habitats used by birds in the northwest portion of Bristol Bay and the adjacent Kilbuck and Ahklun mountains. In the 809 days we were present, we conducted 53 studies or surveys of birds in the region. We gathered information on 185 species, of which 65% (121) nested, 10% (19) probably nested, and 11% (21) were permanent residents in the region. Most breeding or probably breeding forms were of North American (58%; 81) or Beringian (24%; 33) affinity, while the remainder of the species were of Panboreal (17%; 24) and Old World (1%; 2) affinity. Similarly, most of the 44 migrants and visitants were of North American (41%; 18) affinity, while the remainder were of Beringian (32%; 14) and Panboreal (27%; 12) affinity. Of the 140 species that nested or probably nested, 53% (73) were abundant to fairly common, 29% (40) were uncommon to very rare, and 20% (27) were localized. Shrub thicket, dwarf shrub mat, coniferous forest, deciduous forest, mixed deciduous-coniferous forest, and fluviatile water and shoreline habitats supported the greatest diversity of species breeding and suspected of breeding. The highest concentrations of birds occurred in the estuaries of Nanvak, Chagvan, and Goodnews bays during spring and fall migrations and on the coastal and island cliffs during the breeding season.The information presented here provides the basis for range extensions of several species. Our records further clarify the known or probable Alaska breeding ranges of 11 species (fork-tailed storm-petrel, Oceanodroma furcata; double-crested cormorant, Phalacrocorax auritus; red-faced cormorant, Phatacrocorax utile, brant, Branta bernicla; king eider, Somateria spectabilis; white-tailed ptarmigan, Lagopus leucurus; black-bellied plover, Pluvialis squatarola; Pacific golden-plover, Pluvialis fulva; lesser yellowlegs, Tringa flavipes; Say's phoebe, Sayomis saya; and Bohemian waxwing, Bombycilla garrulus). We

Winter distributions of North American Plovers in the Laguna Madre regions of Tamaulipas, Mexico and Texas, USA

USGS Publications Warehouse

Mabee, Todd J.; Plissner, Jonathan H.; Haig, Susan M.; Goossen, J.P.

2001-01-01

To determine the distribution and abundance of wintering plovers in the Laguna Madre of Texas and Tamaulipas, surveys were conducted in December 1997 and February 1998, along a 160 km stretch of barrier islands in Mexico and- 40 km of shoreline on South Padre Island, Texas. Altogether, 5,673 individuals, representing six plover species, were recorded during the surveys. Black-bellied Plovers Pluvialis squatarola were the most numerous (3 ,013 individuals) representing 53% of the total number of plovers observed. Numbers of Piping Charadriusm elodu, Snowy C . alexandrinus, Semipalmated C. semipalmatus and Wilson's Plovers C. wilsonia were 739, 1,345, 561, and 13 birds, respectively. Most individuals (97%) of all species except Wilson's Plovers were observed on bayside flats of the barrier islands. Similar numbers of Piping Plovers were recorded at South Padre Island, Texas, and in the Laguna Madre de Tamaulipas. Over 85% of the individuals of each of the other species were found in the more extensively surveyed Mexico portion of Laguna Madre. In Tamaulipas, most plover species were observed more often on algal flats than any other substrate. These results provide evidence of the value of these systems as wintering areas for plover species and indicate the need for more extensive survey efforts to determine temporal and spatial variation in the distribution of these species within the Laguna ecosystem.

Outdoor cultivation of microalgae for carotenoid production: current state and perspectives.

PubMed

Del Campo, José A; García-González, Mercedes; Guerrero, Miguel G

2007-04-01

Microalgae are a major natural source for a vast array of valuable compounds, including a diversity of pigments, for which these photosynthetic microorganisms represent an almost exclusive biological resource. Yellow, orange, and red carotenoids have an industrial use in food products and cosmetics as vitamin supplements and health food products and as feed additives for poultry, livestock, fish, and crustaceans. The growing worldwide market value of carotenoids is projected to reach over US$1,000 million by the end of the decade. The nutraceutical boom has also integrated carotenoids mainly on the claim of their proven antioxidant properties. Recently established benefits in human health open new uses for some carotenoids, especially lutein, an effective agent for the prevention and treatment of a variety of degenerative diseases. Consumers' demand for natural products favors development of pigments from biological sources, thus increasing opportunities for microalgae. The biotechnology of microalgae has gained considerable progress and relevance in recent decades, with carotenoid production representing one of its most successful domains. In this paper, we review the most relevant features of microalgal biotechnology related to the production of different carotenoids outdoors, with a main focus on beta-carotene from Dunaliella, astaxanthin from Haematococcus, and lutein from chlorophycean strains. We compare the current state of the corresponding production technologies, based on either open-pond systems or closed photobioreactors. The potential of scientific and technological advances for improvements in yield and reduction in production costs for carotenoids from microalgae is also discussed.

Migration stopovers and the conservation of arctic-breeding Calidrine sandpipers

USGS Publications Warehouse

Skagen, Susan K.

2006-01-01

Long-distance migration, one of the most physically demanding events in the animal kingdom, is well developed in many species of Charadriidae and Scolopacidae. Some shorebirds renowned for their extraordinary long-distance migrations, notably American Golden-Plover (Pluvialis dominica), Red Knot (Calidris canutus rufa), and White-rumped Sandpiper (C. fuscicollis), travel as many as 15,000 km between southern South American wintering grounds and Canadian Arctic breeding areas. Migration strategies of shorebirds vary in many aspects. There are remarkable accounts of shorebirds, such as northbound Red Knots, that stage in a few key sites for 2–3 weeks and lay on extensive body stores, then fly nonstop for distances of ≤2,500 km (Harrington 2001, Piersma et al. 2005). Less well known are the examples of populations that refuel only briefly at stopover sites, disperse broadly on the landscape, and fly shorter distances between sites (Skagen 1997, Haig et al. 1998, Warnock et al. 1998). This latter pattern applies to many long-distance migrant shorebirds that cross the interior plains of North America during spring and fall migrations. For them, interior wetland complexes provide critical refueling resources along the direct routes between summering and wintering grounds (Skagen et al. 1999). In this issue of The Auk, Krapu et al. (2006) describe patterns and implications of fat deposition by Semipalmated Sandpipers (C. pusilla), White-rumped Sandpipers, and Baird's Sandpipers (C. bairdii) refueling during northward migration across the prairies of mid-continental North America.

The Effect of Astaxanthin-Rich Microalgae “Haematococcus pluvialis” and Wholemeal Flours Incorporation in Improving the Physical and Functional Properties of Cookies

PubMed Central

Hossain, A. K. M. Mofasser; Brennan, Margaret A.; Mason, Susan L.; Guo, Xinbo; Zeng, Xin An

2017-01-01

Marine-based food supplements can improve human nutrition. In an effort to modulate glycaemic response and enhance nutritional aspects, marine-derived algal food rich in astaxanthin was used in the formulation of a model food (wholemeal cookie). Astaxanthin substitution of cookies made from three flours (wheat, barley and oat) demonstrated a significant reduction in the rate of glucose released during in vitro digestion together with an increase in the total phenolic content (TPC) and antioxidant capacity of the food. The significantly (p < 0.005) lower free glucose release was observed from cookies with 15% astaxanthin, followed by 10% and then 5% astaxanthin in comparison with control cookies of each flour. Total phenolic content, DPPH radical scavenging and Oxygen Radical Absorbance Capacity (ORAC) value also notably increased with increase in astaxanthin content. The results evidence the potential use of microalgae to enhance the bioactive compounds and lower the glycaemic response of wholemeal flour cookie. PMID:28933728

An experimental assessment of vehicle disturbance effects on migratory shorebirds

USGS Publications Warehouse

Tarr, Nathan M.; Simons, T.R.; Pollock, K.H.

2010-01-01

Off-road vehicle (ORV) traffic is one of several forms of disturbance thought to affect shorebirds at migration stopover sites. Attempts to measure disturbance effects on shorebird habitat use and behavior at stopover sites are difficult because ORV disturbance is frequently confounded with habitat and environmental factors. We used a before-after-control-impact experimental design to isolate effects of vehicle disturbance from shorebird responses to environmental and habitat factors. We manipulated disturbance levels within beach closures along South Core Banks, North Carolina, USA, and measured changes in shorebird abundance and location, as well as the activity of one focal species, the sanderling (Calidris alba), within paired control and impact plots. We applied a discrete treatment level of one flee-response-inducing event every 10 minutes on impact plots. We found that disturbance reduced total shorebird and black-bellied plover (Pluvialis squatarola) abundance and reduced relative use of microhabitat zones above the swash zone (wet sand and dry sand) by sanderlings, black-bellied plovers, willets (Tringa semipalmata), and total shorebirds. Sanderlings and total shorebirds increased use of the swash zone in response to vehicle disturbance. Disturbance reduced use of study plots by sanderlings for resting and increased sanderling activity, but we did not detect an effect of vehicle disturbance on sanderling foraging activity. We provide the first estimates of how a discrete level of disturbance affects shorebird distributions among ocean beach microhabitats. Our findings provide a standard to which managers can compare frequency and intensity of disturbance events at other shorebird stopover and roosting sites and indicate that limiting disturbance will contribute to use of a site by migratory shorebirds. ?? 2010 The Wildlife Society.

Is it safe to nest near conspicuous neighbours? Spatial patterns in predation risk associated with the density of American Golden-Plover nests

PubMed Central

Trottier-Paquet, Myriam; Bêty, Joël; Lamarre, Vincent; Lecomte, Nicolas

2016-01-01

Predation is one of the main factors explaining nesting mortality in most bird species. Birds can avoid nest predation or reduce predation pressure by breeding at higher latitude, showing anti-predator behaviour, selecting nest sites protected from predators, and nesting in association with protective species. American Golden-Plovers (Pluvialis dominica) defend their territory by using various warning and distraction behaviours displayed at varying levels of intensity (hereafter “conspicuous behaviour”), as well as more aggressive behaviours such as aerial attacks, but only in some populations. Such antipredator behaviour has the potential to repel predators and thus benefit the neighbouring nests by decreasing their predation risk. Yet, conspicuous behaviour could also attract predators by signalling the presence of a nest. To test for the existence of a protective effect associated with the conspicuous antipredator behaviour of American Golden-Plovers, we studied the influence of proximity to plover nests on predation risk of artificial nests on Igloolik Island (Nunavut, Canada) in July 2014. We predicted that the predation risk of artificial nests would decrease with proximity to and density of plover nests. We monitored 18 plover nests and set 35 artificial nests at 30, 50, 100, 200, and 500 m from seven of those plover nests. We found that the predation risk of artificial nests increases with the density of active plover nests. We also found a significant negative effect of the distance to the nearest active protector nest on predation risk of artificial nests. Understanding how the composition and structure of shorebird communities generate spatial patterns in predation risks represents a key step to better understand the importance of these species of conservation concern in tundra food webs. PMID:27602257

Is it safe to nest near conspicuous neighbours? Spatial patterns in predation risk associated with the density of American Golden-Plover nests.

PubMed

Giroux, Marie-Andrée; Trottier-Paquet, Myriam; Bêty, Joël; Lamarre, Vincent; Lecomte, Nicolas

2016-01-01

Predation is one of the main factors explaining nesting mortality in most bird species. Birds can avoid nest predation or reduce predation pressure by breeding at higher latitude, showing anti-predator behaviour, selecting nest sites protected from predators, and nesting in association with protective species. American Golden-Plovers (Pluvialis dominica) defend their territory by using various warning and distraction behaviours displayed at varying levels of intensity (hereafter "conspicuous behaviour"), as well as more aggressive behaviours such as aerial attacks, but only in some populations. Such antipredator behaviour has the potential to repel predators and thus benefit the neighbouring nests by decreasing their predation risk. Yet, conspicuous behaviour could also attract predators by signalling the presence of a nest. To test for the existence of a protective effect associated with the conspicuous antipredator behaviour of American Golden-Plovers, we studied the influence of proximity to plover nests on predation risk of artificial nests on Igloolik Island (Nunavut, Canada) in July 2014. We predicted that the predation risk of artificial nests would decrease with proximity to and density of plover nests. We monitored 18 plover nests and set 35 artificial nests at 30, 50, 100, 200, and 500 m from seven of those plover nests. We found that the predation risk of artificial nests increases with the density of active plover nests. We also found a significant negative effect of the distance to the nearest active protector nest on predation risk of artificial nests. Understanding how the composition and structure of shorebird communities generate spatial patterns in predation risks represents a key step to better understand the importance of these species of conservation concern in tundra food webs.

Hyaluronidase activity in the salivary glands of tabanid flies.

PubMed

Volfova, Vera; Tothova, Viktorie; Volf, Petr

2016-06-01

Tabanids are haematophagous insects that act as biological and mechanical vectors of various diseases, including viruses, bacteria and parasites. The saliva of these insects contains strong anticoagulant and vasodilatory activities as well as immunoregulatory peptides. Here we demonstrate pronounced hyaluronidase (hyase) activity in ten tabanid species of the genera Chrysops, Haematopota, Hybomitra and Tabanus. Compared to other haematophagous insects, the ability of tabanid hyases to hydrolyze hyaluronic acid (HA) is extremely high, for example the enzyme activity of Hybomitra muehlfeldi was found to be 32-fold higher than the salivary hyase activity of the sand fly Phlebotomus papatasi. Hyases of all ten tabanid species tested also cleaved chondroitin sulfate A, another glycosaminoglycan present in the extracellular matrix of vertebrates. The pH optimum of the enzyme activity was measured in eight tabanid species; the hyase of Haemopota pluvialis was the only one with optimum at pH 4.0, while in the other seven species the activity optimum was at 5.0. SDS PAGE zymography showed the monomeric character of the enzymes in all tabanid species tested. Under non-reducing conditions the activities were visible as single bands with estimated MW between 35 and 52 kDa. The very high hyaluronidase activity in tabanid saliva might be related to their aggressive biting behavior as well as to their high efficiency as mechanical vectors. As they are supposedly involved in the enlargement of feeding hematomas, hyases might contribute to the mechanical transmission of pathogens. Pathogens present in vector mouthparts are co-inoculated into the vertebrate host together with saliva and may benefit from increased tissue permeability and the immunomodulatory activity of the salivary hyase. Copyright © 2016 The Author(s). Published by Elsevier Ltd.. All rights reserved.

Concentrations of 17 elements, including mercury, and their relationship to fitness measures in arctic shorebirds and their eggs.

PubMed

Hargreaves, Anna L; Whiteside, Douglas P; Gilchrist, Grant

2010-07-15

Exposure to contaminants is one hypothesis proposed to explain the global decline in shorebirds, and this is of particular concern in the arctic. However, little information exists on contaminant levels in arctic-breeding shorebirds, especially in Canada. We studied potential contaminants in three biparental shorebird species nesting in Nunavut, Canada: ruddy turnstones (Arenaria interpres), black-bellied plovers (Pluvialis squatarola) and semipalmated plovers (Charadrius semipalmatus). Blood, feathers and eggs were analyzed for As, Be, Cd, Co, Cr, Cu, Fe, Hg, Mn, Mo, Ni, Pb, Sb, Se, Tl, V, and Zn. We assessed whether element concentrations a) differed among species and sexes, b) were correlated among pairs and their eggs, and c) were related to fitness endpoints, namely body condition, blood-parasite load, nest survival days, and hatching success. Non-essential elements were found at lower concentrations than essential elements, with the exception of Hg. Maximum Hg levels in blood approached those associated with toxicological effects in other bird species, but other elements were well below known toxicological thresholds. Reproductive success was negatively correlated with paternal Hg and maternal Pb, although these effects were generally weak and varied among tissues. Element levels were positively correlated within pairs for blood-Hg (turnstones) and feather-Ni and Cr (semipalmated plovers); concentrations in eggs and maternal blood were never correlated. Concentrations of many elements differed among species, but there was no evidence that any species had higher overall exposure to non-essential metals. In conclusion, whereas we found little evidence that exposure to the majority of these elements is leading to declines of the species studied here, Hg levels were of potential concern and both Hg and Pb warrant further monitoring. Copyright 2010 Elsevier B.V. All rights reserved.

Wind effects on prey availability: How northward migrating waders use brackish and hypersaline lagoons in the sivash, Ukraine

NASA Astrophysics Data System (ADS)

Verkuil, Yvonne; Koolhaas, Anita; Van Der Winden, Jan

Large numbers of waders migrating northward in spring use the Sivash, a large system of shallow, brackish and hypersaline lagoons in the Black Sea and Azov Sea region (Ukraine). The bottoms of these lagoons are often uncovered by the wind. Hence, for waders the time and space available for feeding depend on wind conditions. In hypersaline lagoons the benthic and pelagic fauna was very poor, consisting mainly of chironomid larvae (0.19 g AFDM·m -2) and brine shrimps Artemia salina, respectively. Brine shrimp abundance was correlated with salinity, wind force, wind direction and water depth. Dunlin Calidris alpina and curlew sandpiper Calidris ferruginea were the only species feeding on brine shrimp. As brine shrimp densities are higher in deeper water, smaller waders such as broad-billed sandpipers Limicola falcinellus are too short-legged to reach exploitable densities of brine shrimp. In brackish lagoons the benthic and pelagic fauna was rich, consisting of polychaetes, bivalves, gastropods, chironomid larvae, isopods and amphipods (8.9 to 30.5 g AFDM·m -2), but there were no brine shrimps. Prey biomass increased with the distance from the coast, being highest on the site that was most frequently inundated. Dunlin, broad-billed sandpiper and grey plover Pluvialis squatarola were the most abundant birds in the brackish lagoon. Due to the effects of wind-tides only a small area was usually available as a feeding site. Gammarus insensibilis was the alternative prey resource in the water layer, and their density varied with wind direction in the same way as brine shrimp. Curlew sandpipers and dunlins in the hypersaline lagoons and broad-billed sandpipers in the brackish lagoons often changed feeding sites, probably following the variation in prey availability. Only because of the large size and variety of lagoons are waders in the Sivash always able to find good feeding sites.

Concentrations of 17 elements, including mercury, in the tissues, food and abiotic environment of Arctic shorebirds.

PubMed

Hargreaves, Anna L; Whiteside, Douglas P; Gilchrist, Grant

2011-09-01

Exposure to contaminants is one hypothesis proposed to explain the global decline in shorebirds, and is also an increasing concern in the Arctic. We assessed potential contaminants (As, Be, Cd, Co, Cr, Cu, Fe, Hg, Mn, Mo, Ni, Pb, Sb, Se, Tl, V, and Zn) at a shorebird breeding site in Nunavut, Canada. We compared element levels in soil, invertebrates and shorebird blood to assess evidence for bioconcentration and biomagnification within the Arctic-based food chain. We tested whether elements in blood, feathers and eggs of six shorebird species (Pluvialis squatarola, Calidris alpina, C. fuscicollis, Phalaropus fulicarius, Charadrius semipalmatus, and Arenaria interpres) were related to fitness endpoints: adult body condition, blood-parasite load, egg size, eggshell thickness, nest duration, and hatching success. To facilitate comparison to other sites, we summarise the published data on toxic metals in shorebird blood and egg contents. Element concentrations and invertebrate composition differed strongly among habitats, and habitat use and element concentrations differed among shorebird species. Hg, Se, Cd, Cu, and Zn bioconcentrated from soil to invertebrates, and Hg, Se and Fe biomagnified from invertebrates to shorebird blood. As, Ni, Pb, Co and Mn showed significant biodilution from soil to invertebrates to shorebirds. Soil element levels were within Canadian guidelines, and invertebrate Hg levels were below dietary levels suggested for the protection of wildlife. However, maximum Hg in blood and eggs approached levels associated with toxicological effects and Hg-pollution in other bird species. Parental blood-Hg was negatively related to egg volume, although the relationship varied among species. No other elements approached established toxicological thresholds. In conclusion, whereas we found little evidence that exposure to elements at this site is leading to the declines of the species studied, Hg, as found elsewhere in the Canadian Arctic, is of potential

Inventory of montane-nesting birds in the Arctic Network of National Parks, Alaska

USGS Publications Warehouse

Tibbitts, T.L.; Ruthrauff, D.R.; Gill, Robert E.; Handel, Colleen M.

2006-01-01

The Alaska Science Center of the U.S. Geological Survey conducted an inventory of birds in montane areas of the four northern parks in the Arctic Network of National Parks, Alaska. This effort represents the first comprehensive assessment of breeding range and habitat associations for the majority of avian species in the Arctic Network. Ultimately, these data provide a framework upon which to design future monitoring programs.A stratified random sampling design was used to select sample plots (n = 73 plots) that were allocated in proportion to the availability of ecological subsections. Point counts (n = 1,652) were conducted to quantify abundance, distribution, and habitat associations of birds. Field work occurred over three years (2001 to 2003) during two-week-long sessions in late May through early June that coincided with peak courtship activity of breeding birds.Totals of 53 species were recorded in Cape Krusenstern National Monument, 91 in Noatak National Preserve, 57 in Kobuk Valley National Park, and 96 in Gates of the Arctic National Park and Preserve. Substantial proportions of species in individual parks are considered species of conservation concern (18 to 26%) or species of stewardship responsibility of the land managers in the region (8 to 18%). The most commonly detected passerines on point counts included Redpoll spp. (Carduelis flammea and C. hornemanni), Savannah Sparrow (Passerculus sandwichensis), and American Tree Sparrow (Spizella arborea). The most numerous shorebirds were American Golden-Plover (Pluvialis dominica), Wilson’s Snipe (Gallinago delicata), and Whimbrel (Numenius phaeopus). Most species were detected at low rates, reflecting the low breeding densities (and/or low detectabilities) of birds in the montane Arctic. Suites of species were associated with particular ranges of elevation and showed strong associations with particular habitat types.

Inventory of montane-nesting birds in Katmai and Lake Clark national parks and preserves

USGS Publications Warehouse

Ruthrauff, Daniel R.; Tibbitts, Lee; Gill, Robert E.; Handel, Colleen M.

2007-01-01

As part of the National Park Service’s Inventory and Monitoring Program, biologists from the U. S. Geological Survey’s Alaska Science Center conducted an inventory of birds in montane regions of Katmai and Lake Clark National Parks and Preserves during 2004–2006. We used a stratified random survey design to allocate samples by ecological subsection. To survey for birds, we conducted counts at 468 points across 29, 10-km x 10-km (6.2-mi x 6.2-mi) sample plots in Katmai and 417 points across 25, 10-km x 10-km sample plots in Lake Clark. We detected 92 and 104 species in Katmai and Lake Clark, respectively, including 40 species of conservation concern. We detected three species not previously recorded in Katmai (Ring-necked Duck [Aythya collaris], Lesser Scaup [Aythya affinis], and White-tailed Ptarmigan [Lagopus leucurus]) and two species not previously recorded in Lake Clark (Northern Flicker [Colaptes auratus ] and Olive-sided Flycatcher [Contopus cooperi]). The most commonly detected species in both parks was Golden-crowned Sparrow (Zonotrichia atricapilla); Fox Sparrow (Passerella iliaca) and American Pipit (Anthus rubescens) were abundant and widely-distributed as well. We defined sites as low (100–350 m), middle (351–600 m), or high (601–1,620 m) elevation based on the distribution of vegetation cover, and similarly categorized the 34 most-commonly detected species based on the mean elevation of sample points at which they were detected. High elevation (i.e., alpine) sites were characterized by high percent cover of dwarf shrub and bare ground habitat and supported species like Rock Ptarmigan (L. mutus), American Golden-Plover (Pluvialis dominica), Wandering Tattler (Tringa incana), Surfbird (Aphriza virgata), and Snow Bunting (Plectrophenax nivalis), all species of conservation concern. This inventory represents the first systematic survey of birds nesting in montane regions of both parks. Results from this inventory can form the foundation of

Seasonality and daily activity of male and female tabanid flies monitored in a Hungarian hill-country pasture by new polarization traps and traditional canopy traps.

PubMed

Herczeg, Tamás; Blahó, Miklós; Száz, Dénes; Kriska, György; Gyurkovszky, Mónika; Farkas, Róbert; Horváth, Gábor

2014-11-01

Blood-sucking female tabanid flies cause serious problems for animals and humans. For the control of tabanids, the knowledge about their seasonality and daily activity is of great importance. Earlier, only traditional traps capturing exclusively female tabanids have been used to survey tabanid activity. The data of such temporal trapping do not reflect correctly the activity of male and female tabanid flies. Our major aim was to monitor the trapping numbers of male and female tabanids during a 3-month summer survey in Hungary. We used (i) conventional canopy traps with liquid traps on the ground beneath the canopy and (ii) L-shaped sticky traps with vertical and horizontal components. Our other goal was to compare the efficiencies of the two components of each trap type used. We observed two greater peaks of the trapping number of tabanids. These peaks started with increased catches of female tabanids captured by the canopy traps and the vertical sticky traps and ended with a dominance of male and female tabanids caught by the liquid traps and the horizontal sticky traps. The swarming periods were interrupted by rainy/cool days, when the number of tabanids decreased drastically. Among the 17 species, six dominated and composed 89.4% of the captured tabanids: Haematopota pluvialis, Tabanus tergestinus, Tabanus bromius, Tabanus maculicornis, Tabanus bovinus and Atylotus loewianus. The number of water-seeking male and female tabanids rose up to 12-13 h and then decreased but had a secondary peak at about 17 h. The stochastic weather change and the communities of different species resulted in large standard deviations of the averaged number of tabanids in the course of a day. The horizontally polarizing (liquid and horizontal sticky) traps captured both male and female specimens and were about three times more efficient than the canopy and vertical sticky traps that caught only females. The results of the horizontal sticky traps corresponded to those of the liquid

Efficacy of the repellent N,N-diethyl-3-methyl-benzamide (DEET) against tabanid flies on horses evaluated in a field test in Switzerland.

PubMed

Herholz, C; Kopp, C; Wenger, M; Mathis, A; Wägeli, S; Roth, N

2016-05-15

pluvialis, but also non-specified arthropods. The design of the present study simulated practical conditions, allowed to quantify the number of tabanids flies and to demonstrate repellency of DEET in horses. Copyright © 2016 Elsevier B.V. All rights reserved.