Critical evaluation and modeling of algal harvesting using dissolved air flotation. DAF Algal Harvesting Modeling

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

Zhang, Xuezhi; Hewson, John C.; Amendola, Pasquale

In our study, Chlorella zofingiensis harvesting by dissolved air flotation (DAF) was critically evaluated with regard to algal concentration, culture conditions, type and dosage of coagulants, and recycle ratio. Harvesting efficiency increased with coagulant dosage and leveled off at 81%, 86%, 91%, and 87% when chitosan, Al 3+, Fe 3+, and cetyl trimethylammonium bromide (CTAB) were used at dosages of 70, 180, 250, and 500 mg g -1, respectively. The DAF efficiency-coagulant dosage relationship changed with algal culture conditions. In evaluating the influence of the initial algal concentration and recycle ratio revealed that, under conditions typical for algal harvesting, wemore » found that it is possible that the number of bubbles is insufficient. A DAF algal harvesting model was developed to explain this observation by introducing mass-based floc size distributions and a bubble limitation into the white water blanket model. Moreover, the model revealed the importance of coagulation to increase floc-bubble collision and attachment, and the preferential interaction of bubbles with larger flocs, which limited the availability of bubbles to the smaller sized flocs. The harvesting efficiencies predicted by the model agree reasonably with experimental data obtained at different Al 3+ dosages, algal concentrations, and recycle ratios. Based on this modeling, critical parameters for efficient algal harvesting were identified.« less

Accelerating Commercialization of Algal Biofuels Through Partnerships (Brochure)

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

Not Available

2011-10-01

This brochure describes National Renewable Energy Laboratory's (NREL's) algal biofuels research capabilities and partnership opportunities. NREL is accelerating algal biofuels commercialization through: (1) Advances in applied biology; (2) Algal strain development; (3) Development of fuel conversion pathways; (4) Techno-economic analysis; and (5) Development of high-throughput lipid analysis methodologies. NREL scientists and engineers are addressing challenges across the algal biofuels value chain, including algal biology, cultivation, harvesting and extraction, and fuel conversion. Through partnerships, NREL can share knowledge and capabilities in the following areas: (1) Algal Biology - A fundamental understanding of algal biology is key to developing cost-effective algal biofuelsmore » processes. NREL scientists are experts in the isolation and characterization of microalgal species. They are identifying genes and pathways involved in biofuel production. In addition, they have developed a high-throughput, non-destructive technique for assessing lipid production in microalgae. (2) Cultivation - NREL researchers study algal growth capabilities and perform compositional analysis of algal biomass. Laboratory-scale photobioreactors and 1-m2 open raceway ponds in an on-site greenhouse allow for year-round cultivation of algae under a variety of conditions. A bioenergy-focused algal strain collection is being established at NREL, and our laboratory houses a cryopreservation system for long-term maintenance of algal cultures and preservation of intellectual property. (3) Harvesting and Extraction - NREL is investigating cost-effective harvesting and extraction methods suitable for a variety of species and conditions. Areas of expertise include cell wall analysis and deconstruction and identification and utilization of co-products. (4) Fuel Conversion - NREL's excellent capabilities and facilities for biochemical and thermochemical conversion of biomass to biofuels

Plant and algal cell walls: diversity and functionality

PubMed Central

Popper, Zoë A.; Ralet, Marie-Christine; Domozych, David S.

2014-01-01

Background Although plants and many algae (e.g. the Phaeophyceae, brown, and Rhodophyceae, red) are only very distantly related they are united in their possession of carbohydrate-rich cell walls, which are of integral importance being involved in many physiological processes. Furthermore, wall components have applications within food, fuel, pharmaceuticals, fibres (e.g. for textiles and paper) and building materials and have long been an active topic of research. As shown in the 27 papers in this Special Issue, as the major deposit of photosynthetically fixed carbon, and therefore energy investment, cell walls are of undisputed importance to the organisms that possess them, the photosynthetic eukaryotes (plants and algae). The complexities of cell wall components along with their interactions with the biotic and abiotic environment are becoming increasingly revealed. Scope The importance of plant and algal cell walls and their individual components to the function and survival of the organism, and for a number of industrial applications, are illustrated by the breadth of topics covered in this issue, which includes papers concentrating on various plants and algae, developmental stages, organs, cell wall components, and techniques. Although we acknowledge that there are many alternative ways in which the papers could be categorized (and many would fit within several topics), we have organized them as follows: (1) cell wall biosynthesis and remodelling, (2) cell wall diversity, and (3) application of new technologies to cell walls. Finally, we will consider future directions within plant cell wall research. Expansion of the industrial uses of cell walls and potentially novel uses of cell wall components are both avenues likely to direct future research activities. Fundamentally, it is the continued progression from characterization (structure, metabolism, properties and localization) of individual cell wall components through to defining their roles in almost every

Plant and algal cell walls: diversity and functionality.

PubMed

Popper, Zoë A; Ralet, Marie-Christine; Domozych, David S

2014-10-01

Although plants and many algae (e.g. the Phaeophyceae, brown, and Rhodophyceae, red) are only very distantly related they are united in their possession of carbohydrate-rich cell walls, which are of integral importance being involved in many physiological processes. Furthermore,wall components have applications within food, fuel, pharmaceuticals, fibres (e.g. for textiles and paper) and building materials and have long been an active topic of research. As shown in the 27 papers in this Special Issue, as the major deposit of photosynthetically fixed carbon, and therefore energy investment, cell walls are of undisputed importance to the organisms that possess them, the photosynthetic eukaryotes ( plants and algae). The complexities of cell wall components along with their interactions with the biotic and abiotic environment are becoming increasingly revealed. The importance of plant and algal cell walls and their individual components to the function and survival of the organism, and for a number of industrial applications, are illustrated by the breadth of topics covered in this issue, which includes papers concentrating on various plants and algae, developmental stages, organs, cell wall components, and techniques. Although we acknowledge that there are many alternative ways in which the papers could be categorized (and many would fit within several topics), we have organized them as follows: (1) cell wall biosynthesis and remodelling, (2) cell wall diversity, and (3) application of new technologies to cell walls. Finally, we will consider future directions within plant cell wall research. Expansion of the industrial uses of cell walls and potentially novel uses of cell wall components are both avenues likely to direct future research activities. Fundamentally, it is the continued progression from characterization (structure, metabolism, properties and localization) of individual cell wall components through to defining their roles in almost every aspect of plant

Prevalent and persistent viral infection in cultures of the coral algal endosymbiont Symbiodinium

NASA Astrophysics Data System (ADS)

Weynberg, Karen D.; Neave, Matthew; Clode, Peta L.; Voolstra, Christian R.; Brownlee, Christopher; Laffy, Patrick; Webster, Nicole S.; Levin, Rachel A.; Wood-Charlson, Elisha M.; van Oppen, Madeleine J. H.

2017-09-01

Reef corals are under threat from bleaching and disease outbreaks that target both the host animal and the algal symbionts within the coral holobiont. A viral origin for coral bleaching has been hypothesized, but direct evidence has remained elusive. Using a multifaceted approach incorporating flow cytometry, transmission electron microscopy, DNA and RNA virome sequencing, we show that type C1 Symbiodinium cultures host a nucleocytoplasmic large double-stranded DNA virus (NCLDV) related to Phycodnaviridae and Mimiviridae, a novel filamentous virus of unknown phylogenetic affiliation, and a single-stranded RNA virus related to retroviruses. We discuss implications of these findings for laboratory-based experiments using Symbiodinium cultures.

Biomass recycle as a means to improve the energy efficiency of CELSS algal culture systems

NASA Technical Reports Server (NTRS)

Radmer, R.; Cox, J.; Lieberman, D.; Behrens, P.; Arnett, K.

1987-01-01

Algal cultures can be very rapid and efficient means to generate biomass and regenerate the atmosphere for closed environmental life support systems. However, as in the case of most higher plants, a significant fraction of the biomass produced by most algae cannot be directly converted to a useful food product by standard food technology procedures. This waste biomass will serve as an energy drain on the overall system unless it can be efficiently recycled without a significant loss of its energy content. Experiments are reported in which cultures of the alga Scenedesmus obliquus were grown in the light and at the expense of an added carbon source, which either replaced or supplemented the actinic light. As part of these experiments, hydrolyzed waste biomass from these same algae were tested to determine whether the algae themselves could be made part of the biological recycling process. Results indicate that hydrolyzed algal (and plant) biomass can serve as carbon and energy sources for the growth of these algae, suggesting that the efficiency of the closed system could be significantly improved using this recycling process.

Dynamic metabolic exchange governs a marine algal-bacterial interaction.

PubMed

Segev, Einat; Wyche, Thomas P; Kim, Ki Hyun; Petersen, Jörn; Ellebrandt, Claire; Vlamakis, Hera; Barteneva, Natasha; Paulson, Joseph N; Chai, Liraz; Clardy, Jon; Kolter, Roberto

2016-11-18

Emiliania huxleyi is a model coccolithophore micro-alga that generates vast blooms in the ocean. Bacteria are not considered among the major factors influencing coccolithophore physiology. Here we show through a laboratory model system that the bacterium Phaeobacter inhibens , a well-studied member of the Roseobacter group, intimately interacts with E. huxleyi. While attached to the algal cell, bacteria initially promote algal growth but ultimately kill their algal host. Both algal growth enhancement and algal death are driven by the bacterially-produced phytohormone indole-3-acetic acid. Bacterial production of indole-3-acetic acid and attachment to algae are significantly increased by tryptophan, which is exuded from the algal cell. Algal death triggered by bacteria involves activation of pathways unique to oxidative stress response and programmed cell death. Our observations suggest that bacteria greatly influence the physiology and metabolism of E. huxleyi. Coccolithophore-bacteria interactions should be further studied in the environment to determine whether they impact micro-algal population dynamics on a global scale.

Host–microbe interactions as a driver of acclimation to salinity gradients in brown algal cultures

PubMed Central

Dittami, Simon M; Duboscq-Bidot, Laëtitia; Perennou, Morgan; Gobet, Angélique; Corre, Erwan; Boyen, Catherine; Tonon, Thierry

2016-01-01

Like most eukaryotes, brown algae live in association with bacterial communities that frequently have beneficial effects on their development. Ectocarpus is a genus of small filamentous brown algae, which comprises a strain that has recently colonized freshwater, a rare transition in this lineage. We generated an inventory of bacteria in Ectocarpus cultures and examined the effect they have on acclimation to an environmental change, that is, the transition from seawater to freshwater medium. Our results demonstrate that Ectocarpus depends on bacteria for this transition: cultures that have been deprived of their associated microbiome do not survive a transfer to freshwater, but restoring their microflora also restores the capacity to acclimate to this change. Furthermore, the transition between the two culture media strongly affects the bacterial community composition. Examining a range of other closely related algal strains, we observed that the presence of two bacterial operational taxonomic units correlated significantly with an increase in low salinity tolerance of the algal culture. Despite differences in the community composition, no indications were found for functional differences in the bacterial metagenomes predicted to be associated with algae in the salinities tested, suggesting functional redundancy in the associated bacterial community. Our study provides an example of how microbial communities may impact the acclimation and physiological response of algae to different environments, and thus possibly act as facilitators of speciation. It paves the way for functional examinations of the underlying host–microbe interactions, both in controlled laboratory and natural conditions. PMID:26114888

Application of a fluorometric microplate algal toxicity assay for riverine periphytic algal species.

PubMed

Nagai, Takashi; Taya, Kiyoshi; Annoh, Hirochica; Ishihara, Satoru

2013-08-01

Although riverine periphytic algae attached to riverbed gravel are dominant species in flowing rivers, there is limited toxicity data on them because of the difficulty in cell culture and assays. Moreover, it is well known that sensitivity to pesticides differ markedly among species, and therefore the toxicity data for multiple species need to be efficiently obtained. In this study, we investigated the use of fluorometric microplate toxicity assay for testing periphytic algal species. We selected five candidate test algal species Desmodesmus subspicatus, Achnanthidium minutissimum, Navicula pelliculosa, Nitzschia palea, and Pseudanabaena galeata. The selected species are dominant in the river, include a wide range of taxon, and represent actual species composition. Other additional species were also used to compare the sensitivity and suitability of the microplate assay. A 96-well microplate was used as a test chamber and algal growth was measured by in-vivo fluorescence. Assay conditions using microplate and fluorometric measurement were established, and sensitivities of 3,5-dichlorophenol as a reference substance were assayed. The 50 percent effect concentrations (EC50s) obtained by fluorometric microplate assay and those obtained by conventional Erlenmeyer flask assay conducted in this study were consistent. Moreover, the EC50 values of 3,5-dichlorophenol were within the reported confidence intervals in literature. These results supported the validity of our microplate assay. Species sensitivity distribution (SSD) analysis was conducted using the EC50s of five species. The SSD was found to be similar to the SSD obtained using additional tested species, suggesting that SSD using the five species largely represents algal sensitivity. Our results provide a useful and efficient method for high-tier probabilistic ecological risk assessment of pesticides. Copyright © 2013 Elsevier Inc. All rights reserved.

Experimental study on the interspecific interactions between the two bloom-forming algal species and the rotifer Brachionus plicatilis

NASA Astrophysics Data System (ADS)

Xie, Zhihao; Xiao, Hui; Tang, Xuexi; Cai, Hengjiang

2009-06-01

The interspecific interactions between the rotifer Brachionus plicatilis and two harmful algal blooms (HAB) species were investigated experimentally by single culture method. B. plicatilis population and the growth of the two algae were compared at different algal cell densities. The results demonstrated that the B. plicatilis obtained sufficient nutrition from Prorocentrum donghaiense to support net population increase. With exposure to 2.5×104 cells mL-1 of P. donghaiense, the number of B. plicatilis increased faster than it did when exposed to other four algal densities (5, 10, 15 and 20 ×104 cells mL-1), and the increase rate of B. plicatilis population ( r) at this algal density was 0.104 ± 0.015 rd-1. Cell densities of P. donghaiense decreased due to the grazing of B. plicatilis. In contrast, Heterosigma akashiwo had an adverse effect on B. plicatilis population and its growth was largely unaffected by rotifer grazing. In this case, B. plicatilis population decreased and H. akashiwo grew at a rate similar to that of the control.

Dynamic metabolic exchange governs a marine algal-bacterial interaction

PubMed Central

Segev, Einat; Wyche, Thomas P; Kim, Ki Hyun; Petersen, Jörn; Ellebrandt, Claire; Vlamakis, Hera; Barteneva, Natasha; Paulson, Joseph N; Chai, Liraz; Clardy, Jon; Kolter, Roberto

2016-01-01

Emiliania huxleyi is a model coccolithophore micro-alga that generates vast blooms in the ocean. Bacteria are not considered among the major factors influencing coccolithophore physiology. Here we show through a laboratory model system that the bacterium Phaeobacter inhibens, a well-studied member of the Roseobacter group, intimately interacts with E. huxleyi. While attached to the algal cell, bacteria initially promote algal growth but ultimately kill their algal host. Both algal growth enhancement and algal death are driven by the bacterially-produced phytohormone indole-3-acetic acid. Bacterial production of indole-3-acetic acid and attachment to algae are significantly increased by tryptophan, which is exuded from the algal cell. Algal death triggered by bacteria involves activation of pathways unique to oxidative stress response and programmed cell death. Our observations suggest that bacteria greatly influence the physiology and metabolism of E. huxleyi. Coccolithophore-bacteria interactions should be further studied in the environment to determine whether they impact micro-algal population dynamics on a global scale. DOI: http://dx.doi.org/10.7554/eLife.17473.001 PMID:27855786

Enhanced Production of Green Tide Algal Biomass through Additional Carbon Supply

PubMed Central

de Paula Silva, Pedro H.; Paul, Nicholas A.; de Nys, Rocky; Mata, Leonardo

2013-01-01

Intensive algal cultivation usually requires a high flux of dissolved inorganic carbon (Ci) to support productivity, particularly for high density algal cultures. Carbon dioxide (CO2) enrichment can be used to overcome Ci limitation and enhance productivity of algae in intensive culture, however, it is unclear whether algal species with the ability to utilise bicarbonate (HCO3 −) as a carbon source for photosynthesis will benefit from CO2 enrichment. This study quantified the HCO3 − affinity of three green tide algal species, Cladophora coelothrix, Cladophora patentiramea and Chaetomorpha linum, targeted for biomass and bioenergy production. Subsequently, we quantified productivity and carbon, nitrogen and ash content in response to CO2 enrichment. All three species had similar high pH compensation points (9.7–9.9), and grew at similar rates up to pH 9, demonstrating HCO3 − utilization. Algal cultures enriched with CO2 as a carbon source had 30% more total Ci available, supplying twenty five times more CO2 than the control. This higher Ci significantly enhanced the productivity of Cladophora coelothrix (26%), Chaetomorpha linum (24%) and to a lesser extent for Cladophora patentiramea (11%), compared to controls. We demonstrated that supplying carbon as CO2 can enhance the productivity of targeted green tide algal species under intensive culture, despite their clear ability to utilise HCO3 −. PMID:24324672

Enhanced production of green tide algal biomass through additional carbon supply.

PubMed

de Paula Silva, Pedro H; Paul, Nicholas A; de Nys, Rocky; Mata, Leonardo

2013-01-01

Intensive algal cultivation usually requires a high flux of dissolved inorganic carbon (Ci) to support productivity, particularly for high density algal cultures. Carbon dioxide (CO2) enrichment can be used to overcome Ci limitation and enhance productivity of algae in intensive culture, however, it is unclear whether algal species with the ability to utilise bicarbonate (HCO3 (-)) as a carbon source for photosynthesis will benefit from CO2 enrichment. This study quantified the HCO3 (-) affinity of three green tide algal species, Cladophora coelothrix, Cladophora patentiramea and Chaetomorpha linum, targeted for biomass and bioenergy production. Subsequently, we quantified productivity and carbon, nitrogen and ash content in response to CO2 enrichment. All three species had similar high pH compensation points (9.7-9.9), and grew at similar rates up to pH 9, demonstrating HCO3 (-) utilization. Algal cultures enriched with CO2 as a carbon source had 30% more total Ci available, supplying twenty five times more CO2 than the control. This higher Ci significantly enhanced the productivity of Cladophora coelothrix (26%), Chaetomorpha linum (24%) and to a lesser extent for Cladophora patentiramea (11%), compared to controls. We demonstrated that supplying carbon as CO2 can enhance the productivity of targeted green tide algal species under intensive culture, despite their clear ability to utilise HCO3 (-).

Culturing Selenastrum capricornutum (Chlorophyta) in a synthetic algal nutrient medium with defined mineral particulates

USGS Publications Warehouse

Kuwabara, J.S.; Davis, J.A.; Chang, Cecily C.Y.

1985-01-01

Algal nutrient studies in chemically-defined media typically employ a synthetic chelator to prevent iron hydroxide precipitation. Micronutrient-particulate interactions may, however, significantly affect chemical speciation and hence biovailability of these nutrients in natural waters. A technique is described by which Selenastrum capricornutum Printz (Chlorophyta) may be cultured in a medium where trace metal speciation (except iron) is controlled, not by organic chelation, but by sorption onto titanium dioxide. Application of this culturing protocol in conjunction with results from sorption studies of nutrient ions on mineral particles provides a means of studying biological impacts of sorptive processes in aquatic environments. ?? 1985 Dr W. Junk Publishers.

Maintenance of algal endosymbionts in Paramecium bursaria: a simple model based on population dynamics.

PubMed

Iwai, Sosuke; Fujiwara, Kenji; Tamura, Takuro

2016-09-01

Algal endosymbiosis is widely distributed in eukaryotes including many protists and metazoans, and plays important roles in aquatic ecosystems, combining phagotrophy and phototrophy. To maintain a stable symbiotic relationship, endosymbiont population size in the host must be properly regulated and maintained at a constant level; however, the mechanisms underlying the maintenance of algal endosymbionts are still largely unknown. Here we investigate the population dynamics of the unicellular ciliate Paramecium bursaria and its Chlorella-like algal endosymbiont under various experimental conditions in a simple culture system. Our results suggest that endosymbiont population size in P. bursaria was not regulated by active processes such as cell division coupling between the two organisms, or partitioning of the endosymbionts at host cell division. Regardless, endosymbiont population size was eventually adjusted to a nearly constant level once cells were grown with light and nutrients. To explain this apparent regulation of population size, we propose a simple mechanism based on the different growth properties (specifically the nutrient requirements) of the two organisms, and based from this develop a mathematical model to describe the population dynamics of host and endosymbiont. The proposed mechanism and model may provide a basis for understanding the maintenance of algal endosymbionts. © 2015 Society for Applied Microbiology and John Wiley & Sons Ltd.

Characterization of Amoeboaphelidium protococcarum, an Algal Parasite New to the Cryptomycota Isolated from an Outdoor Algal Pond Used for the Production of Biofuel

PubMed Central

Letcher, Peter M.; Lopez, Salvador; Schmieder, Robert; Lee, Philip A.; Behnke, Craig; Powell, Martha J.; McBride, Robert C.

2013-01-01

Mass culture of algae for the production of biofuels is a developing technology designed to offset the depletion of fossil fuel reserves. However, large scale culture of algae in open ponds can be challenging because of incidences of infestation with algal parasites. Without knowledge of the identity of the specific parasite and how to control these pests, algal-based biofuel production will be limited. We have characterized a eukaryotic parasite of Scenedesmus dimorphus growing in outdoor ponds used for biofuel production. We demonstrated that as the genomic DNA of parasite FD01 increases, the concentration of S. dimorphus cells decreases; consequently, this is a highly destructive pathogen. Techniques for culture of the parasite and host were developed, and the endoparasite was identified as the Aphelidea, Amoeboaphelidium protococcarum. Phylogenetic analysis of ribosomal sequences revealed that parasite FD01 placed within the recently described Cryptomycota, a poorly known phylum based on two species of Rozella and environmental samples. Transmission electron microscopy demonstrated that aplanospores of the parasite produced filose pseudopodia, which contained fine fibers the diameter of actin microfilaments. Multiple lipid globules clustered and were associated with microbodies, mitochondria and a membrane cisternae, an arrangement characteristic of the microbody-lipid globule complex of chytrid zoospores. After encystment and attachment to the host cells, the parasite injected its protoplast into the host between the host cell wall and plasma membrane. At maturity the unwalled parasite occupied the entire host cell. After cleavage of the protoplast into aplanospores, a vacuole and lipids remained in the host cell. Amoeboaphelidium protococcarum isolate FD01 is characteristic of the original description of this species and is different from strain X-5 recently characterized. Our results help put a face on the Cryptomycota, revealing that the phylum is more

Algal autolysate medium to label proteins for NMR in mammalian cells.

PubMed

Fuccio, Carmelo; Luchinat, Enrico; Barbieri, Letizia; Neri, Sara; Fragai, Marco

2016-04-01

In-cell NMR provides structural and functional information on proteins directly inside living cells. At present, the high costs of the labeled media for mammalian cells represent a limiting factor for the development of this methodology. Here we report a protocol to prepare a homemade growth medium from Spirulina platensis autolysate, suitable to express uniformly labeled proteins inside mammalian cells at a reduced cost-per-sample. The human proteins SOD1 and Mia40 were overexpressed in human cells grown in (15)N-enriched S. platensis algal-derived medium, and high quality in-cell NMR spectra were obtained.

Coupled nutrient removal and biomass production with mixed algal culture: impact of biotic and abiotic factors.

PubMed

Su, Yanyan; Mennerich, Artur; Urban, Brigitte

2012-08-01

The influence of biotic (algal inoculum concentration) and abiotic factors (illumination cycle, mixing velocity and nutrient strength) on the treatment efficiency, biomass generation and settleability were investigated with selected mixed algal culture. Dark condition led to poor nutrient removal efficiency. No significant difference in the N, P removal and biomass settleability between continuous and alternating illumination was observed, but a higher biomass generation capability for the continuous illumination was obtained. Different mixing velocity led to similar phosphorus removal efficiencies (above 98%) with different retention times. The reactor with 300 rpm mixing velocity had the best N removal capability. For the low strength wastewater, the N rates were 5.4±0.2, 9.1±0.3 and 10.8±0.3 mg/l/d and P removal rates were 0.57±0.03, 0.56±0.03 and 0.72±0.05 mg/l/d for reactors with the algal inoculum concentration of 0.2, 0.5 and 0.8 g/l, respectively. Low nutrient removal efficiency and poor biomass settleability were obtained for high strength wastewater. Copyright © 2012 Elsevier Ltd. All rights reserved.

Constraints to commercialization of algal fuels.

PubMed

Chisti, Yusuf

2013-09-10

Production of algal crude oil has been achieved in various pilot scale facilities, but whether algal fuels can be produced in sufficient quantity to meaningfully displace petroleum fuels, has been largely overlooked. Limitations to commercialization of algal fuels need to be understood and addressed for any future commercialization. This review identifies the major constraints to commercialization of transport fuels from microalgae. Algae derived fuels are expensive compared to petroleum derived fuels, but this could change. Unfortunately, improved economics of production are not sufficient for an environmentally sustainable production, or its large scale feasibility. A low-cost point supply of concentrated carbon dioxide colocated with the other essential resources is necessary for producing algal fuels. An insufficiency of concentrated carbon dioxide is actually a major impediment to any substantial production of algal fuels. Sustainability of production requires the development of an ability to almost fully recycle the phosphorous and nitrogen nutrients that are necessary for algae culture. Development of a nitrogen biofixation ability to support production of algal fuels ought to be an important long term objective. At sufficiently large scale, a limited supply of freshwater will pose a significant limitation to production even if marine algae are used. Processes for recovering energy from the algal biomass left after the extraction of oil, are required for achieving a net positive energy balance in the algal fuel oil. The near term outlook for widespread use of algal fuels appears bleak, but fuels for niche applications such as in aviation may be likely in the medium term. Genetic and metabolic engineering of microalgae to boost production of fuel oil and ease its recovery, are essential for commercialization of algal fuels. Algae will need to be genetically modified for improved photosynthetic efficiency in the long term. Copyright © 2013 Elsevier B.V. All

Role of gas vesicles and intra-colony spaces during the process of algal bloom formation.

PubMed

Zhang, Yongsheng; Zheng, Binghui; Jiang, Xia; Zheng, Hao

2013-06-01

Aggregation morphology, vertical distribution, and algal density were analyzed during the algal cell floating process in three environments. The role of gas vesicles and intra-colony spaces was distinguished by algal blooms treated with ultrasonic waves and high pressure. Results demonstrated that the two buoyancy providers jointly provide buoyancy for floating algal cells. The results were also confirmed by force analysis. In the simulation experiment, the buoyancy acting on algal cells was greater than its gravity at sample ports 2 and 3 of a columnar-cultivated cell vessel, and intra-colony spaces were not detected. In Taihu Lake, gas vesicle buoyancy was notably less than total algal cell gravity. Buoyancy provided by intra-colony spaces exceeded total algal cell gravity at the water surface, but not at other water depths. In the Daning River, total buoyancies provided by the two buoyancy providers were less than total algal cell gravity at different water depths.

Transcriptome analysis illuminates the nature of the intracellular interaction in a vertebrate-algal symbiosis

PubMed Central

Burns, John A; Zhang, Huanjia; Hill, Elizabeth; Kim, Eunsoo; Kerney, Ryan

2017-01-01

During embryonic development, cells of the green alga Oophila amblystomatis enter cells of the salamander Ambystoma maculatum forming an endosymbiosis. Here, using de novo dual-RNA seq, we compared the host salamander cells that harbored intracellular algae to those without algae and the algae inside the animal cells to those in the egg capsule. This two-by-two-way analysis revealed that intracellular algae exhibit hallmarks of cellular stress and undergo a striking metabolic shift from oxidative metabolism to fermentation. Culturing experiments with the alga showed that host glutamine may be utilized by the algal endosymbiont as a primary nitrogen source. Transcriptional changes in salamander cells suggest an innate immune response to the alga, with potential attenuation of NF-κB, and metabolic alterations indicative of modulation of insulin sensitivity. In stark contrast to its algal endosymbiont, the salamander cells did not exhibit major stress responses, suggesting that the host cell experience is neutral or beneficial. DOI: http://dx.doi.org/10.7554/eLife.22054.001 PMID:28462779

Municipal wastewater treatment and biomass accumulation with a wastewater-born and settleable algal-bacterial culture.

PubMed

Su, Yanyan; Mennerich, Artur; Urban, Brigitte

2011-05-01

A wastewater-born and settleable algal-bacterial culture, cultivated in a stirred tank photobioreactor under lab conditions, was used to remove the carbon and nutrients in municipal wastewater and accumulate biomass simultaneously. The algal-bacterial culture showed good settleable property and could totally settle down over 20 min, resulting in a reduction of total suspended solids from an initial 1.84 to 0.016 g/l. The average removal efficiencies of chemical oxygen demand, total kjeldahl nitrogen and phosphate were 98.2 ± 1.3%, 88.3 ± 1.6% and 64.8 ± 1.0% within 8 days, respectively, while the average biomass productivity was 10.9 ± 1.1 g/m(2) · d. Accumulation into biomass, identified as the main nitrogen and phosphorus removal mechanism, accounted for 44.9 ± 0.4% and 61.6 ± 0.5% of total inlet nitrogen and phosphorus, respectively. Microscopic analysis showed the main algae species in the bioreactor were filamentous blue-green algae. Furthermore, denaturing gradient gel electrophoresis and 16S rDNA gene sequencing revealed that the main bacteria present in the photobioreactor were consortia with sequences similar to those of Flavobacteria, Gammaproteobacteria, Bacteroidia and Betaproteobacteria. This study explores a better understanding of an algae-bacteria system and offers new information on further usage of biomass accumulated during treatment. Copyright © 2011 Elsevier Ltd. All rights reserved.

Analysis-Software for Hyperspectral Algal Reflectance Probes v. 1.0

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

Timlin, Jerilyn A.; Reichardt, Thomas A.; Jenson, Travis J.

This software provides onsite analysis of the hyperspectral reflectance data acquired on an outdoor algal pond by a multichannel, fiber-coupled spectroradiometer. The analysis algorithm is based on numerical inversion of a reflectance model, in which the above-water reflectance is expressed as a function of the single backscattering albedo, which is dependent on the backscatter and absorption coefficients of the algal culture, which are in turn related to the algal biomass and pigment optical activity, respectively. Prior to the development of this software, while raw multichannel data were displayed in real time, analysis required a post-processing procedure to extract the relevantmore » parameters. This software provides the capability to track the temporal variation of such culture parameters in real time, as raw data are being acquired, or can be run in a post processing mode. The software allows the user to select between different algal species, incorporate the appropriate calibration data, and observe the quality of the resulting model inversions.« less

Luminescent Solar Concentrators in the Algal Industry

NASA Astrophysics Data System (ADS)

Hellier, Katie; Corrado, Carley; Carter, Sue; Detweiler, Angela; Bebout, Leslie

2013-03-01

Today's industry for renewable energy sources and highly efficient energy management systems is rapidly increasing. Development of increased efficiency Luminescent Solar Concentrators (LSCs) has brought about new applications for commercial interests, including greenhouses for agricultural crops. This project is taking first steps to explore the potential of LSCs to enhance production and reduce costs for algae and cyanobacteria used in biofuels and nutraceuticals. This pilot phase uses LSC filtered light for algal growth trials in greenhouses and laboratory experiments, creating specific wavelength combinations to determine effects of discrete solar light regimes on algal growth and the reduction of heating and water loss in the system. Enhancing the optimal spectra for specific algae will not only increase production, but has the potential to lessen contamination of large scale production due to competition from other algae and bacteria. Providing LSC filtered light will reduce evaporation and heating in regions with limited water supply, while the increased energy output from photovoltaic cells will reduce costs of heating and mixing cultures, thus creating a more efficient and cost effective production system.

Fueling Future with Algal Genomics

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

Grigoriev, Igor

Algae constitute a major component of fundamental eukaryotic diversity, play profound roles in the carbon cycle, and are prominent candidates for biofuel production. The US Department of Energy Joint Genome Institute (JGI) is leading the world in algal genome sequencing (http://jgi.doe.gov/Algae) and contributes of the algal genome projects worldwide (GOLD database, 2012). The sequenced algal genomes offer catalogs of genes, networks, and pathways. The sequenced first of its kind genomes of a haptophyte E.huxleyii, chlorarachniophyte B.natans, and cryptophyte G.theta fill the gaps in the eukaryotic tree of life and carry unique genes and pathways as well as molecular fossils ofmore » secondary endosymbiosis. Natural adaptation to conditions critical for industrial production is encoded in algal genomes, for example, growth of A.anophagefferens at very high cell densities during the harmful algae blooms or a global distribution across diverse environments of E.huxleyii, able to live on sparse nutrients due to its expanded pan-genome. Communications and signaling pathways can be derived from simple symbiotic systems like lichens or complex marine algae metagenomes. Collectively these datasets derived from algal genomics contribute to building a comprehensive parts list essential for algal biofuel development.« less

Vernalophrys algivore gen. nov., sp. nov. (Rhizaria: Cercozoa: Vampyrellida), a New Algal Predator Isolated from Outdoor Mass Culture of Scenedesmus dimorphus

PubMed Central

Patterson, David J.; Li, Yunguang; Hu, Zixuan; Sommerfeld, Milton; Chen, Yongsheng

2015-01-01

Microbial contamination is the main cause of loss of biomass yield in microalgal cultures, especially under outdoor environmental conditions. Little is known about the identities of microbial contaminants in outdoor mass algal cultures. In this study, a new genus and species of vampyrellid amoeba, Vernalophrys algivore, is described from cultures of Scenedesmus dimorphus in open raceway ponds and outdoor flat-panel photobioreactors. This vampyrellid amoeba was a significant grazer of Scenedesmus and was frequently associated with a very rapid decline in algal numbers. We report on the morphology, subcellular structure, feeding behavior, molecular phylogeny, and life cycle. The new amoeba resembles Leptophrys in the shape of trophozoites and pseudopodia and in the mechanism of feeding (mainly by engulfment). It possesses two distinctive regions in helix E10_1 (nucleotides 117 to 119, CAA) and E23_1 (nucleotides 522 and 523, AG) of the 18S rRNA gene. It did not form a monophyletic group with Leptophrys in molecular phylogenetic trees. We establish a new genus, Vernalophrys, with the type species Vernalophrys algivore. The occurrence, impact of the amoeba on mass culture of S. dimorphus, and means to reduce vampyrellid amoeba contamination in Scenedesmus cultures are addressed. The information obtained from this study will be useful for developing an early warning system and control measures for preventing or treating this contaminant in microalgal mass cultures. PMID:25819973

Change in Photosystem II Photochemistry During Algal Growth Phases of Chlorella vulgaris and Scenedesmus obliquus.

PubMed

Oukarroum, Abdallah

2016-06-01

Sensitivity of photosynthetic processes towards environmental stress is used as a bioanalytical tool to evaluate the responses of aquatic plants to a changing environment. In this paper, change of biomass density, chlorophyll a fluorescence and photosynthetic parameters during growth phases of two microalgae Chlorella vulgaris and Scenedesmus obliquus were studied. The photosynthetic growth behaviour changed significantly with cell age and algae species. During the exponential phase of growth, the photosynthesis capacity reached its maximum and decreased in ageing algal culture during stationary phase. In conclusion, the chlorophyll a fluorescence OJIP method and the derived fluorescence parameters would be an accurate method for obtaining information on maximum photosynthetic capacities and monitoring algal cell growth. This will contribute to more understanding, for example, of toxic actions of pollutants in microalgae test.

Resolving Mixed Algal Species in Hyperspectral Images

PubMed Central

Mehrubeoglu, Mehrube; Teng, Ming Y.; Zimba, Paul V.

2014-01-01

We investigated a lab-based hyperspectral imaging system's response from pure (single) and mixed (two) algal cultures containing known algae types and volumetric combinations to characterize the system's performance. The spectral response to volumetric changes in single and combinations of algal mixtures with known ratios were tested. Constrained linear spectral unmixing was applied to extract the algal content of the mixtures based on abundances that produced the lowest root mean square error. Percent prediction error was computed as the difference between actual percent volumetric content and abundances at minimum RMS error. Best prediction errors were computed as 0.4%, 0.4% and 6.3% for the mixed spectra from three independent experiments. The worst prediction errors were found as 5.6%, 5.4% and 13.4% for the same order of experiments. Additionally, Beer-Lambert's law was utilized to relate transmittance to different volumes of pure algal suspensions demonstrating linear logarithmic trends for optical property measurements. PMID:24451451

Effects of ozone and peroxone on algal separation via dispersed air flotation.

PubMed

Nguyen, Truc Linh; Lee, D J; Chang, J S; Liu, J C

2013-05-01

Effects of pre-oxidation on algal separation by dispersed air flotation were examined. Ozone (O3) and peroxone (O3 and H2O2) could induce cell lysis, release of intracellular organic matter (IOM), and mineralization of organic substances. Separation efficiency of algal cells improved when pre-oxidized. Total of 76.4% algal cells was separated at 40 mg/L of N-cetyl-N-N-N-trimethylammonium bromide (CTAB), while 95% were separated after 30-min ozonation. Pre-oxidation by ozone and peroxone also enhanced flotation separation efficiency of dissolved organic carbon (DOC), polysaccharide, and protein, in which peroxone process exerted more significantly than O3. Two main mechanisms were involved in flotation separation of unoxidized algal suspension, namely hydrophobic cell surface and cell flocculation resulting from CTAB adsorption. However, flocculation by CTAB was hindered for pre-oxidized algal suspensions. It implied that the compositional changes in extracellular organic matter (EOM) by pre-oxidation were more determined for flotation separation of pre-oxidized cells. Copyright © 2012 Elsevier B.V. All rights reserved.

Investigating why recycling gravity harvested algae increases harvestability and productivity in high rate algal ponds.

PubMed

Park, J B K; Craggs, R J; Shilton, A N

2013-09-15

It has previously been shown that recycling gravity harvested algae promotes Pediastrum boryanum dominance and improves harvestability and biomass production in pilot-scale High Rate Algal Ponds (HRAPs) treating domestic wastewater. In order to confirm the reproducibility of these findings and investigate the mechanisms responsible, this study utilized twelve 20 L outdoor HRAP mesocosms operated with and without algal recycling. It then compared the recycling of separated solid and liquid components of the harvested biomass against un-separated biomass. The work confirmed that algal recycling promoted P. boryanum dominance, improved 1 h-settleability by >20% and increased biomass productivity by >25% compared with controls that had no recycling. With regard to the improved harvestability, of particular interest was that recycling the liquid fraction alone caused a similar improvement in settleability as recycling the solid fraction. This may be due to the presence of extracellular polymeric substances in the liquid fraction. While there are many possible mechanisms that could account for the increased productivity with algal recycling, all but two were systematically eliminated: (i) the mean cell residence time was extended thereby increasing the algal concentration and more fully utilizing the incident sunlight and, (ii) the relative proportions of algal growth stages (which have different specific growth rates) was changed, resulting in a net increase in the overall growth rate of the culture. Copyright © 2013 Elsevier Ltd. All rights reserved.

Production of biofuel using molluscan pseudofeces derived from algal cells

DOEpatents

Das, Keshav C.; Chinnasamy, Senthil; Shelton, James; Wilde, Susan B.; Haynie, Rebecca S.; Herrin, James A.

2012-08-28

Embodiments of the present disclosure provide for novel strategies to harvest algal lipids using mollusks which after feeding algae from the growth medium can convert algal lipids into their biomass or excrete lipids in their pseudofeces which makes algae harvesting energy efficient and cost effective. The bioconverter, filter-feeding mollusks and their pseudofeces can be harvested and converted to biocrude using an advanced thermochemical liquefaction technology. Methods, systems, and materials are disclosed for the harvest and isolation of algal lipids from the mollusks, molluscan feces and molluscan pseudofeces.

Vernalophrys algivore gen. nov., sp. nov. (Rhizaria: Cercozoa: Vampyrellida), a New Algal Predator Isolated from Outdoor Mass Culture of Scenedesmus dimorphus.

PubMed

Gong, Yingchun; Patterson, David J; Li, Yunguang; Hu, Zixuan; Sommerfeld, Milton; Chen, Yongsheng; Hu, Qiang

2015-06-15

Microbial contamination is the main cause of loss of biomass yield in microalgal cultures, especially under outdoor environmental conditions. Little is known about the identities of microbial contaminants in outdoor mass algal cultures. In this study, a new genus and species of vampyrellid amoeba, Vernalophrys algivore, is described from cultures of Scenedesmus dimorphus in open raceway ponds and outdoor flat-panel photobioreactors. This vampyrellid amoeba was a significant grazer of Scenedesmus and was frequently associated with a very rapid decline in algal numbers. We report on the morphology, subcellular structure, feeding behavior, molecular phylogeny, and life cycle. The new amoeba resembles Leptophrys in the shape of trophozoites and pseudopodia and in the mechanism of feeding (mainly by engulfment). It possesses two distinctive regions in helix E10_1 (nucleotides 117 to 119, CAA) and E23_1 (nucleotides 522 and 523, AG) of the 18S rRNA gene. It did not form a monophyletic group with Leptophrys in molecular phylogenetic trees. We establish a new genus, Vernalophrys, with the type species Vernalophrys algivore. The occurrence, impact of the amoeba on mass culture of S. dimorphus, and means to reduce vampyrellid amoeba contamination in Scenedesmus cultures are addressed. The information obtained from this study will be useful for developing an early warning system and control measures for preventing or treating this contaminant in microalgal mass cultures. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

A new approach for the estimation of phytoplankton cell counts associated with algal blooms.

PubMed

Nazeer, Majid; Wong, Man Sing; Nichol, Janet Elizabeth

2017-07-15

This study proposes a method for estimating phytoplankton cell counts associated with an algal bloom, using satellite images coincident with in situ and meteorological parameters. Satellite images from Landsat Thematic Mapper (TM), Enhanced Thematic Mapper Plus (ETM+), Operational Land Imager (OLI) and HJ-1 A/B Charge Couple Device (CCD) sensors were integrated with the meteorological observations to provide an estimate of phytoplankton cell counts. All images were atmospherically corrected using the Second Simulation of the Satellite Signal in the Solar Spectrum (6S) atmospheric correction method with a possible error of 1.2%, 2.6%, 1.4% and 2.3% for blue (450-520nm), green (520-600nm), red (630-690nm) and near infrared (NIR 760-900nm) wavelengths, respectively. Results showed that the developed Artificial Neural Network (ANN) model yields a correlation coefficient (R) of 0.95 with the in situ validation data with Sum of Squared Error (SSE) of 0.34cell/ml, Mean Relative Error (MRE) of 0.154cells/ml and a bias of -504.87. The integration of the meteorological parameters with remote sensing observations provided a promising estimation of the algal scum as compared to previous studies. The applicability of the ANN model was tested over Hong Kong as well as over Lake Kasumigaura, Japan and Lake Okeechobee, Florida USA, where algal blooms were also reported. Further, a 40-year (1975-2014) red tide occurrence map was developed and revealed that the eastern and southern waters of Hong Kong are more vulnerable to red tides. Over the 40 years, 66% of red tide incidents were associated with the Dinoflagellates group, while the remainder were associated with the Diatom group (14%) and several other minor groups (20%). The developed technology can be applied to other similar environments in an efficient and cost-saving manner. Copyright © 2017 Elsevier B.V. All rights reserved.

Molecular identification and nanoremediation of microbial contaminants in algal systems using untreated wastewater.

PubMed

Limayem, Alya; Gonzalez, Francisco; Micciche, Andrew; Haller, Edward; Nayak, Bina; Mohapatra, Shyam

2016-12-01

Wastewater-algal biomass is a promising option to biofuel production. However, microbial contaminants constitute a substantial barrier to algal biofuel yield. A series of algal strains, Nannochloris oculata and Chlorella vulgaris samples (n = 30), were purchased from the University of Texas, and were used for both stock flask cultures and flat-panel vertical bioreactors. A number of media were used for isolation and differentiation of potential contaminants according to laboratory standards (CLSI). Conventional PCR amplification was performed followed by 16S rDNA sequencing to identify isolates at the species level. Nanotherapeutics involving a nanomicellar combination of natural chitosan and zinc oxide (CZNPs) were tested against the microbial lytic groups through Minimum Inhibitory Concentration (MIC) tests and Transmission Electronic Microscopy (TEM). Results indicated the presence of Pseudomonas spp., Bacillus pumilus/ safensis, Cellulosimicrobium cellulans, Micrococcus luteus and Staphylococcus epidermidis strains at a substantial level in the wastewater-fed algal reactors. TEM confirmed the effectiveness of CZNPs on the lytic group while the average MICs (mg/mL) detected for the strains, Pseudomonas spp, Micrococcus luteus, and Bacillus pumilus were 0.417, 3.33, and 1.458, respectively. Conclusively, CZNP antimicrobials proved to be effective as inhibitory agents against currently identified lytic microbial group, did not impact algae cells, and shows promise for in situ interventions.

Algal endosymbionts in European Hydra strains reflect multiple origins of the zoochlorella symbiosis.

PubMed

Rajević, Nives; Kovačević, Goran; Kalafatić, Mirjana; Gould, Sven B; Martin, William F; Franjević, Damjan

2015-12-01

Symbiotic associations are of broad significance in evolution and biodiversity. Green Hydra is a classic example of endosymbiosis. In its gastrodermal myoepithelial cells it harbors endosymbiotic unicellular green algae, most commonly from the genus Chlorella. We reconstructed the phylogeny of cultured algal endosymbionts isolated and maintained in laboratory conditions for years from green Hydra strains collected from four different geographical sites within Croatia, one from Germany and one from Israel. Nuclear (18S rDNA, ITS region) and chloroplast markers (16S, rbcL) for maximum likelihood phylogenetic analyses were used. We focused on investigating the positions of these algal endosymbiotic strains within the chlorophyte lineage. Molecular analyses established that different genera and species of unicellular green algae are present as endosymbionts in green Hydra, showing that endosymbiotic algae growing within green Hydra sampled from four Croatian localities are not monophyletic. Our results indicate that the intracellular algal endosymbionts of green Hydra have become established several times independently in evolution. Copyright © 2015 Elsevier Inc. All rights reserved.

Mollusc-algal chloroplast endosymbiosis. Photosynthesis, thylakoid protein maintenance, and chloroplast gene expression continue for many months in the absence of the algal nucleus.

PubMed

Green, B J; Li, W Y; Manhart, J R; Fox, T C; Summer, E J; Kennedy, R A; Pierce, S K; Rumpho, M E

2000-09-01

Early in its life cycle, the marine mollusc Elysia chlorotica Gould forms an intracellular endosymbiotic association with chloroplasts of the chromophytic alga Vaucheria litorea C. Agardh. As a result, the dark green sea slug can be sustained in culture solely by photoautotrophic CO(2) fixation for at least 9 months if provided with only light and a source of CO(2). Here we demonstrate that the sea slug symbiont chloroplasts maintain photosynthetic oxygen evolution and electron transport activity through photosystems I and II for several months in the absence of any external algal food supply. This activity is correlated to the maintenance of functional levels of chloroplast-encoded photosystem proteins, due in part at least to de novo protein synthesis of chloroplast proteins in the sea slug. Levels of at least one putative algal nuclear encoded protein, a light-harvesting complex protein homolog, were also maintained throughout the 9-month culture period. The chloroplast genome of V. litorea was found to be 119.1 kb, similar to that of other chromophytic algae. Southern analysis and polymerase chain reaction did not detect an algal nuclear genome in the slug, in agreement with earlier microscopic observations. Therefore, the maintenance of photosynthetic activity in the captured chloroplasts is regulated solely by the algal chloroplast and animal nuclear genomes.

The death mechanism of the harmful algal bloom species Alexandrium tamarense induced by algicidal bacterium Deinococcus sp. Y35

PubMed Central

Li, Yi; Zhu, Hong; Lei, Xueqian; Zhang, Huajun; Cai, Guanjing; Chen, Zhangran; Fu, Lijun; Xu, Hong; Zheng, Tianling

2015-01-01

Harmful algal blooms (HABs) cause a variety of deleterious effects on aquatic ecosystems, especially the toxic dinoflagellate Alexandrium tamarense, which poses a serious threat to marine economic and human health based on releasing paralytic shellfish poison into the environment. The algicidal bacterium Deinococcus sp. Y35 which can induce growth inhibition on A. tamarense was used to investigate the functional mechanism. The growth status, reactive oxygen species (ROS) content, photosynthetic system and the nuclear system of algal cells were determined under algicidal activity. A culture of strain Y35 not only induced overproduction of ROS in algal cells within only 0.5 h of treatment, also decrease the total protein content as well as the response of the antioxidant enzyme. Meanwhile, lipid peroxidation was induced and cell membrane integrity was lost. Photosynthetic pigments including chlorophyll a and carotenoid decreased along with the photosynthetic efficiency being significantly inhibited. At the same time, photosynthesis-related gene expression showed down-regulation. More than, the destruction of cell nuclear structure and inhibition of proliferating cell nuclear antigen (PCNA) related gene expression were confirmed. The potential functional mechanism of the algicidal bacterium on A. tamarense was investigated and provided a novel viewpoint which could be used in HABs control. PMID:26441921

The death mechanism of the harmful algal bloom species Alexandrium tamarense induced by algicidal bacterium Deinococcus sp. Y35.

PubMed

Li, Yi; Zhu, Hong; Lei, Xueqian; Zhang, Huajun; Cai, Guanjing; Chen, Zhangran; Fu, Lijun; Xu, Hong; Zheng, Tianling

2015-01-01

Harmful algal blooms (HABs) cause a variety of deleterious effects on aquatic ecosystems, especially the toxic dinoflagellate Alexandrium tamarense, which poses a serious threat to marine economic and human health based on releasing paralytic shellfish poison into the environment. The algicidal bacterium Deinococcus sp. Y35 which can induce growth inhibition on A. tamarense was used to investigate the functional mechanism. The growth status, reactive oxygen species (ROS) content, photosynthetic system and the nuclear system of algal cells were determined under algicidal activity. A culture of strain Y35 not only induced overproduction of ROS in algal cells within only 0.5 h of treatment, also decrease the total protein content as well as the response of the antioxidant enzyme. Meanwhile, lipid peroxidation was induced and cell membrane integrity was lost. Photosynthetic pigments including chlorophyll a and carotenoid decreased along with the photosynthetic efficiency being significantly inhibited. At the same time, photosynthesis-related gene expression showed down-regulation. More than, the destruction of cell nuclear structure and inhibition of proliferating cell nuclear antigen (PCNA) related gene expression were confirmed. The potential functional mechanism of the algicidal bacterium on A. tamarense was investigated and provided a novel viewpoint which could be used in HABs control.

Cell Division Synchronization

DTIC Science & Technology

The report summarizes the progress in the design and construction of automatic equipment for synchronizing cell division in culture by periodic...Concurrent experiments in hypothermic synchronization of algal cell division are reported.

Algal Cell Factories: Approaches, Applications, and Potentials.

PubMed

Fu, Weiqi; Chaiboonchoe, Amphun; Khraiwesh, Basel; Nelson, David R; Al-Khairy, Dina; Mystikou, Alexandra; Alzahmi, Amnah; Salehi-Ashtiani, Kourosh

2016-12-13

With the advent of modern biotechnology, microorganisms from diverse lineages have been used to produce bio-based feedstocks and bioactive compounds. Many of these compounds are currently commodities of interest, in a variety of markets and their utility warrants investigation into improving their production through strain development. In this review, we address the issue of strain improvement in a group of organisms with strong potential to be productive "cell factories": the photosynthetic microalgae. Microalgae are a diverse group of phytoplankton, involving polyphyletic lineage such as green algae and diatoms that are commonly used in the industry. The photosynthetic microalgae have been under intense investigation recently for their ability to produce commercial compounds using only light, CO₂, and basic nutrients. However, their strain improvement is still a relatively recent area of work that is under development. Importantly, it is only through appropriate engineering methods that we may see the full biotechnological potential of microalgae come to fruition. Thus, in this review, we address past and present endeavors towards the aim of creating productive algal cell factories and describe possible advantageous future directions for the field.

Basic and Applied Algal Life Support System Research on Board the Deep Space Gateway

NASA Astrophysics Data System (ADS)

Niederwieser, T.; Zea, L.; Anthony, J.; Stodieck, L.

2018-02-01

We study the effect of long-term preservation methods on DNA damage of algal cultures for BLSS applications. In a secondary step, the Deep Space Gateway serves as a technology demonstration platform for algal photobioreactors in intermittently occupied habitats.

Mechanical algal disruption for efficient biodiesel extraction

NASA Astrophysics Data System (ADS)

Krehbiel, Joel David

Biodiesel from algae provides several benefits over current biodiesel feedstocks, but the energy requirements of processing algae into a useable fuel are currently so high as to be prohibitive. One route to improving this is via disruption of the cells prior to lipid extraction, which can significantly increase energy recovery. Unfortunately, several obvious disruption techniques require more energy than can be gained. This dissertation examines the use of microbubbles to improve mechanical disruption of algal cells using experimental, theoretical, and computational methods. New laboratory experiments show that effective ultrasonic disruption of algae is achieved by adding microbubbles to an algal solution. The configuration studied flows the solution through a tube and insonifies a small section with a high-pressure ultrasound wave. Previous biomedical research has shown effective cell membrane damage on animal cells with similar methods, but the present research is the first to extend such study to algal cells. Results indicate that disruption increases with peak negative pressure between 1.90 and 3.07 MPa and with microbubble concentration up to 12.5 x 107 bubbles/ml. Energy estimates of this process suggest that it requires only one-fourth the currently most-efficient laboratory-scale disruption process. Estimates of the radius near each bubble that causes disruption (i.e. the disruption radius) suggest that it increases with peak negative pressure and is near 9--20 microm for all cases tested. It is anticipated that these procedures can be designed for better efficiency and efficacy, which will be facilitated by identifying the root mechanisms of the bubble-induced disruption. We therefore examine whether bubble expansion alone creates sufficient cell deformation for cell rupture. The spherically-symmetric Marmottant model for bubble dynamics allows estimation of the flow regime under experimental conditions. Bubble expansion is modeled as a point source of

Titanium dioxide nanoparticle exposure reduces algal biomass and alters algal assemblage composition in wastewater effluent-dominated stream mesocosms.

PubMed

Wright, Moncie V; Matson, Cole W; Baker, Leanne F; Castellon, Benjamin T; Watkins, Preston S; King, Ryan S

2018-06-01

A 5-week mesocosm experiment was conducted to investigate the toxicity of titanium dioxide nanoparticles (TiO 2 NPs) to periphytic algae in an environmentally-realistic scenario. We used outdoor experimental streams to simulate the characteristics of central Texas streams receiving large discharges of wastewater treatment plant effluent during prolonged periods of drought. The streams were continually dosed and maintained at two concentrations. The first represents an environmentally relevant concentration of 0.05 mg L -1 (low concentration). The second treatment of 5 mg L -1 (high concentration) was selected to represent a scenario where TiO 2 NPs are used for photocatalytic degradation of pharmaceuticals in wastewater. Algal cell density, chlorophyll-a, ash-free dry mass, algal assemblage composition, and Ti accumulation were determined for the periphyton in the riffle sections of each stream. The high concentration treatment of TiO 2 NPs significantly decreased algal cell density, ash-free dry mass, and chlorophyll-a, and altered algal assemblage composition. Decreased abundance of three typically pollution-sensitive taxa and increased abundance of two genera associated with heavy metal sorption and organic pollution significantly contributed to algal assemblage composition changes in response to TiO 2 NPs. Benefits of the use of TiO 2 NPs in wastewater treatment plants will need to be carefully weighed against the demonstrated ability of these NPs to cause large changes in periphyton that would likely propagate significant effects throughout the stream ecosystem, even in the absence of direct toxicity to higher trophic level organisms. Copyright © 2018 Elsevier B.V. All rights reserved.

Disruption of Cell-to-Cell Signaling Does Not Abolish the Antagonism of Phaeobacter gallaeciensis toward the Fish Pathogen Vibrio anguillarum in Algal Systems

PubMed Central

Prol García, M. J.; D'Alvise, P. W.

2013-01-01

Quorum sensing (QS) regulates Phaeobacter gallaeciensis antagonism in broth systems; however, we demonstrate here that QS is not important for antagonism in algal cultures. QS mutants reduced Vibrio anguillarum to the same extent as the wild type. Consequently, a combination of probiotic Phaeobacter and QS inhibitors is a feasible strategy for aquaculture disease control. PMID:23811510

The Suppressive Activity of Fucofuroeckol-A Derived from Brown Algal Ecklonia stolonifera Okamura on UVB-Induced Mast Cell Degranulation

PubMed Central

Vo, Thanh Sang; Kim, Se-Kwon; Ngo, Dai Hung; Yoon, Na-Young; Bach, Long Giang; Hang, Nguyen Thi Nhat; Ngo, Dai Nghiep

2018-01-01

UV light, especially UVB, is known as a trigger of allergic reaction, leading to mast cell degranulation and histamine release. In this study, phlorotannin Fucofuroeckol-A (F-A) derived from brown algal Ecklonia stolonifera Okamura was evaluated for its protective capability against UVB-induced allergic reaction in RBL-2H3 mast cells. It was revealed that F-A significantly suppress mast cell degranulation via decreasing histamine release as well as intracellular Ca2+ elevation at the concentration of 50 μM. Moreover, the inhibitory effect of F-A on IL-1β and TNF-α productions was also evidenced. Notably, the protective activity of F-A against mast cell degranulation was found due to scavenging ROS production. Accordingly, F-A from brown algal E. stolonifera was suggested to be promising candidate for its protective capability against UVB-induced allergic reaction. PMID:29300311

Mollusc-Algal Chloroplast Endosymbiosis. Photosynthesis, Thylakoid Protein Maintenance, and Chloroplast Gene Expression Continue for Many Months in the Absence of the Algal Nucleus1

PubMed Central

Green, Brian J.; Li, Wei-Ye; Manhart, James R.; Fox, Theodore C.; Summer, Elizabeth J.; Kennedy, Robert A.; Pierce, Sidney K.; Rumpho, Mary E.

2000-01-01

Early in its life cycle, the marine mollusc Elysia chlorotica Gould forms an intracellular endosymbiotic association with chloroplasts of the chromophytic alga Vaucheria litorea C. Agardh. As a result, the dark green sea slug can be sustained in culture solely by photoautotrophic CO2 fixation for at least 9 months if provided with only light and a source of CO2. Here we demonstrate that the sea slug symbiont chloroplasts maintain photosynthetic oxygen evolution and electron transport activity through photosystems I and II for several months in the absence of any external algal food supply. This activity is correlated to the maintenance of functional levels of chloroplast-encoded photosystem proteins, due in part at least to de novo protein synthesis of chloroplast proteins in the sea slug. Levels of at least one putative algal nuclear encoded protein, a light-harvesting complex protein homolog, were also maintained throughout the 9-month culture period. The chloroplast genome of V. litorea was found to be 119.1 kb, similar to that of other chromophytic algae. Southern analysis and polymerase chain reaction did not detect an algal nuclear genome in the slug, in agreement with earlier microscopic observations. Therefore, the maintenance of photosynthetic activity in the captured chloroplasts is regulated solely by the algal chloroplast and animal nuclear genomes. PMID:10982447

Recent Advances in Algal Genetic Tool Development

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

R. Dahlin, Lukas; T. Guarnieri, Michael

The goal of achieving cost-effective biofuels and bioproducts derived from algal biomass will require improvements along the entire value chain, including identification of robust, high-productivity strains and development of advanced genetic tools. Though there have been modest advances in development of genetic systems for the model alga Chlamydomonas reinhardtii, progress in development of algal genetic tools, especially as applied to non-model algae, has generally lagged behind that of more commonly utilized laboratory and industrial microbes. This is in part due to the complex organellar structure of algae, including robust cell walls and intricate compartmentalization of target loci, as well asmore » prevalent gene silencing mechanisms, which hinder facile utilization of conventional genetic engineering tools and methodologies. However, recent progress in global tool development has opened the door for implementation of strain-engineering strategies in industrially-relevant algal strains. Here, we review recent advances in algal genetic tool development and applications in eukaryotic microalgae.« less

Recent Advances in Algal Genetic Tool Development

DOE PAGES

R. Dahlin, Lukas; T. Guarnieri, Michael

2016-06-24

The goal of achieving cost-effective biofuels and bioproducts derived from algal biomass will require improvements along the entire value chain, including identification of robust, high-productivity strains and development of advanced genetic tools. Though there have been modest advances in development of genetic systems for the model alga Chlamydomonas reinhardtii, progress in development of algal genetic tools, especially as applied to non-model algae, has generally lagged behind that of more commonly utilized laboratory and industrial microbes. This is in part due to the complex organellar structure of algae, including robust cell walls and intricate compartmentalization of target loci, as well asmore » prevalent gene silencing mechanisms, which hinder facile utilization of conventional genetic engineering tools and methodologies. However, recent progress in global tool development has opened the door for implementation of strain-engineering strategies in industrially-relevant algal strains. Here, we review recent advances in algal genetic tool development and applications in eukaryotic microalgae.« less

A novel single-parameter approach for forecasting algal blooms.

PubMed

Xiao, Xi; He, Junyu; Huang, Haomin; Miller, Todd R; Christakos, George; Reichwaldt, Elke S; Ghadouani, Anas; Lin, Shengpan; Xu, Xinhua; Shi, Jiyan

2017-01-01

Harmful algal blooms frequently occur globally, and forecasting could constitute an essential proactive strategy for bloom control. To decrease the cost of aquatic environmental monitoring and increase the accuracy of bloom forecasting, a novel single-parameter approach combining wavelet analysis with artificial neural networks (WNN) was developed and verified based on daily online monitoring datasets of algal density in the Siling Reservoir, China and Lake Winnebago, U.S.A. Firstly, a detailed modeling process was illustrated using the forecasting of cyanobacterial cell density in the Chinese reservoir as an example. Three WNN models occupying various prediction time intervals were optimized through model training using an early stopped training approach. All models performed well in fitting historical data and predicting the dynamics of cyanobacterial cell density, with the best model predicting cyanobacteria density one-day ahead (r = 0.986 and mean absolute error = 0.103 × 10 4  cells mL -1 ). Secondly, the potential of this novel approach was further confirmed by the precise predictions of algal biomass dynamics measured as chl a in both study sites, demonstrating its high performance in forecasting algal blooms, including cyanobacteria as well as other blooming species. Thirdly, the WNN model was compared to current algal forecasting methods (i.e. artificial neural networks, autoregressive integrated moving average model), and was found to be more accurate. In addition, the application of this novel single-parameter approach is cost effective as it requires only a buoy-mounted fluorescent probe, which is merely a fraction (∼15%) of the cost of a typical auto-monitoring system. As such, the newly developed approach presents a promising and cost-effective tool for the future prediction and management of harmful algal blooms. Copyright © 2016 Elsevier Ltd. All rights reserved.

Heterotrophic cultivation of Chlorella pyrenoidosa using sucrose as the sole carbon source by co-culture with Rhodotorula glutinis.

PubMed

Wang, Shikai; Wu, Yong; Wang, Xu

2016-11-01

Heterotrophic cultivation of microalgae is a feasible alternative strategy to avoid the light limitation of photoautotrophic culture, but the heterotrophic utilization of disaccharides is difficult for microalgae. Aimed at this problem, a co-culture system was developed by mix culture of C. pyrenoidosa and R. glutinis using sucrose as the sole carbon source. In this system, C. pyrenoidosa could utilize glucose and fructose which were hydrolyzed from sucrose by R. glutinis. The highest specific growth rate and final cell number proportion of algae was 1.02day(-1) and 45%, respectively, when cultured at the initial algal cell number proportion of 95.24% and the final algal cell density was 111.48×10(6)cells/mL. In addition, the lipid content was also promoted due to the synergistic effects in mix culture. This study provides a novel approach using sucrose-riched wastes for the heterotrophic culture of microalgae and may effectively decrease the cost of carbon source. Copyright © 2016 Elsevier Ltd. All rights reserved.

Collection and conversion of algal lipid

NASA Astrophysics Data System (ADS)

Lin, Ching-Chieh

Sustainable economic activities mandate a significant replacement of fossil energy by renewable forms. Algae-derived biofuels are increasingly seen as an alternative source of energy with potential to supplement the world's ever increasing demand. Our primary objective is, once the algae were cultivated, to eliminate or make more efficient energy-intensive processing steps of collection, drying, grinding, and solvent extraction prior to conversion. To overcome the processing barrier, we propose to streamline from cultivated algae to biodiesel via algal biomass collection by sand filtration, cell rupturing with ozone, and immediate transesterification. To collect the algal biomass, the specific Chlorococcum aquaticum suspension was acidified to pH 3.3 to promote agglomeration prior to sand filtration. The algae-loaded filter bed was drained of free water and added with methanol and ozonated for 2 min to rupture cell membrane to accelerate release of the cellular contents. The methanol solution now containing the dissolved lipid product was collected by draining, while the filter bed was regenerated by further ozonation when needed. The results showed 95% collection of the algal biomass from the suspension and a 16% yield of lipid from the algae, as well as restoration of filtration velocity of the sand bed via ozonation. The results further showed increased lipid yield upon cell rupturing and transesterified products composed entirely of fatty acid methyl ester (FAME) compounds, demonstrating that the rupture and transesterification processes could proceed consecutively in the same medium, requiring no separate steps of drying, extraction, and conversion. The FAME products from algae without exposure to ozone were mainly of 16 to 18 carbons containing up to 3 double bonds, while those from algae having been ozonated were smaller, highly saturated hydrocarbons. The new technique streamlines individual steps from cultivated algal lipid to transesterified products and

Free ammonia offers algal crop protection from predators in dairy wastewater and ammonium-rich media.

PubMed

Thomas, Patrick K; Dunn, Gary P; Passero, Maxine; Feris, Kevin P

2017-11-01

Cost-effective methods for protecting crops from grazing organisms like rotifers are needed to reduce the risk of pond crashes in mass algal cultures. We present a novel strategy to optimize the exposure time to free ammonia, via control of media pH, in both defined media and dairy anaerobic digester effluent to suppress rotifers and maintain algal productivity. We tested five different free ammonia exposure times (0, 1, 2, 6, and 12h) and found a significant nonlinear effect of exposure time (p<0.0001) but not pH (p>0.9) on rotifer survival. In both media types, 6-12h of elevated free ammonia significantly reduced Brachionus plicatilis rotifer survival with no negative effects on Nannochloropsis oculata, while shorter exposure times were insufficient to inhibit rotifers, leading to severe algal culture crashes. These results suggest that algal crops can be protected from rotifers, without productivity loss, by elevating free ammonia for 6 or more hours. Copyright © 2017 Elsevier Ltd. All rights reserved.

Increasing algal photosynthetic productivity by integrating ecophysiology with systems biology.

PubMed

Peers, Graham

2014-11-01

Oxygenic photosynthesis is the process by which plants, algae, and cyanobacteria convert sunlight and CO2 into chemical energy and biomass. Previously published estimates suggest that algal photosynthesis is, at best, able to convert approximately 5-7% of incident light energy to biomass and there is opportunity for improvement. Recent analyses of in situ photophysiology in mass cultures of algae and cyanobacteria show that cultivation methods can have detrimental effects on a cell's photophysiology - reinforcing the need to understand the complex responses of cell biology to a highly variable environment. A systems-based approach to understanding the stresses and efficiencies associated with light-energy harvesting, CO2 fixation, and carbon partitioning will be necessary to make major headway toward improving photosynthetic yields. Copyright © 2014 Elsevier Ltd. All rights reserved.

Molluscan cells in culture: primary cell cultures and cell lines

PubMed Central

Yoshino, T. P.; Bickham, U.; Bayne, C. J.

2013-01-01

In vitro cell culture systems from molluscs have significantly contributed to our basic understanding of complex physiological processes occurring within or between tissue-specific cells, yielding information unattainable using intact animal models. In vitro cultures of neuronal cells from gastropods show how simplified cell models can inform our understanding of complex networks in intact organisms. Primary cell cultures from marine and freshwater bivalve and gastropod species are used as biomonitors for environmental contaminants, as models for gene transfer technologies, and for studies of innate immunity and neoplastic disease. Despite efforts to isolate proliferative cell lines from molluscs, the snail Biomphalaria glabrata Say, 1818 embryonic (Bge) cell line is the only existing cell line originating from any molluscan species. Taking an organ systems approach, this review summarizes efforts to establish molluscan cell cultures and describes the varied applications of primary cell cultures in research. Because of the unique status of the Bge cell line, an account is presented of the establishment of this cell line, and of how these cells have contributed to our understanding of snail host-parasite interactions. Finally, we detail the difficulties commonly encountered in efforts to establish cell lines from molluscs and discuss how these difficulties might be overcome. PMID:24198436

Wastewater treatment high rate algal ponds (WWT HRAP) for low-cost biofuel production.

PubMed

Mehrabadi, Abbas; Craggs, Rupert; Farid, Mohammed M

2015-05-01

Growing energy demand and water consumption have increased concerns about energy security and efficient wastewater treatment and reuse. Wastewater treatment high rate algal ponds (WWT HRAPs) are a promising technology that could help solve these challenges concurrently where climate is favorable. WWT HRAPs have great potential for biofuel production as a by-product of WWT, since the costs of algal cultivation and harvest for biofuel production are covered by the wastewater treatment function. Generally, 800-1400 GJ/ha/year energy (average biomass energy content: 20 GJ/ton; HRAP biomass productivity: 40-70 tons/ha/year) can be produced in the form of harvestable biomass from WWT HRAP which can be used to provide community-level energy supply. In this paper the benefits of WWT HRAPs are compared with conventional mass algal culture systems. Moreover, parameters to effectively increase algal energy content and overall energy production from WWT HRAP are discussed including selection of appropriate algal biomass biofuel conversion pathways. Copyright © 2014 Elsevier Ltd. All rights reserved.

Process for selection of oxygen-tolerant algal mutants that produce H{sub 2}

DOEpatents

Ghirardi, M.L.; Seibert, M.

1999-02-16

A process for selection of oxygen-tolerant, H{sub 2}-producing algal mutant cells comprises: (a) growing algal cells photoautotrophically under fluorescent light to mid log phase; (b) inducing algal cells grown photoautotrophically under fluorescent light to mid log phase in step (a) anaerobically by (1) resuspending the cells in a buffer solution and making said suspension anaerobic with an inert gas and (2) incubating the suspension in the absence of light at ambient temperature; (c) treating the cells from step (b) with metronidazole, sodium azide, and added oxygen to controlled concentrations in the presence of white light; (d) washing off metronidazole and sodium azide to obtain final cell suspension; (e) plating said final cell suspension on a minimal medium and incubating in light at a temperature sufficient to enable colonies to appear; (f) counting the number of colonies to determine the percent of mutant survivors; and (g) testing survivors to identify oxygen-tolerant H{sub 2}-producing mutants. 5 figs.

Process for selection of Oxygen-tolerant algal mutants that produce H.sub.2

DOEpatents

Ghirardi, Maria L.; Seibert, Michael

1999-01-01

A process for selection of oxygen-tolerant, H.sub.2 -producing algal mutant cells comprising: (a) growing algal cells photoautotrophically under fluorescent light to mid log phase; (b) inducing algal cells grown photoautrophically under fluorescent light to mid log phase in step (a) anaerobically by (1) resuspending the cells in a buffer solution and making said suspension anaerobic with an inert gas; (2) incubating the suspension in the absence of light at ambient temperature; (c) treating the cells from step (b) with metronidazole, sodium azide, and added oxygen to controlled concentrations in the presence of white light. (d) washing off metronidazole and sodium azide to obtain final cell suspension; (e) plating said final cell suspension on a minimal medium and incubating in light at a temperature sufficient to enable colonies to appear; (f) counting the number of colonies to determine the percent of mutant survivors; and (g) testing survivors to identify oxygen-tolerant H.sub.2 -producing mutants.

Selective algicidal action of peptides against harmful algal bloom species.

PubMed

Park, Seong-Cheol; Lee, Jong-Kook; Kim, Si Wouk; Park, Yoonkyung

2011-01-01

Recently, harmful algal bloom (HAB), also termed "red tide", has been recognized as a serious problem in marine environments according to climate changes worldwide. Many novel materials or methods to prevent HAB have not yet been employed except for clay dispersion, in which can the resulting sedimentation on the seafloor can also cause alteration in marine ecology or secondary environmental pollution. In the current study, we investigated that antimicrobial peptide have a potential in controlling HAB without cytotoxicity to harmless marine organisms. Here, antimicrobial peptides are proposed as new algicidal compounds in combating HAB cells. HPA3 and HPA3NT3 peptides which exert potent antimicrobial activity via pore forming action in plasma membrane showed that HPA3NT3 reduced the motility of algal cells, disrupted their plasma membrane, and induced the efflux of intracellular components. Against raphidoflagellate such as Heterosigma akashiwo, Chattonella sp., and C. marina, it displayed a rapid lysing action in cell membranes at 1~4 µM within 2 min. Comparatively, its lysing effects occurred at 8 µM within 1 h in dinoflagellate such as Cochlodium polykrikoides, Prorocentrum micans, and P. minimum. Moreover, its lysing action induced the lysis of chloroplasts and loss of chlorophyll a. In the contrary, this peptide was not effective against Skeletonema costatum, harmless algal cell, even at 256 µM, moreover, it killed only H. akashiwo or C. marina in co-cultivation with S. costatum, indicating to its selective algicidal activity between harmful and harmless algal cells. The peptide was non-hemolytic against red blood cells of Sebastes schlegeli, the black rockfish, at 120 µM. HAB cells were quickly and selectively lysed following treatment of antimicrobial peptides without cytotoxicity to harmless marine organisms. Thus, the antibiotic peptides examined in our study appear to have much potential in effectively controlling HAB with minimal impact on marine

Loss of phototaxis and degeneration of an eyespot in long-term algal cultures: evidence from ultrastructure and behaviour in the dinoflagellate Kryptoperidinium foliaceum.

PubMed

Moldrup, Morten; Moestrup, Øjvind; Hansen, Per Juel

2013-01-01

Phototaxis provides phytoplankton with the means to orient themselves in a light gradient. This is accomplished using an eyespot and associated organelles. For the dinoflagellate Kryptoperidinium foliaceum, which has been described as having one of the most elaborate eyespot complexes known, positive phototaxis has hitherto not been reported. In this study, we show that a newly isolated strain of K. foliaceum is indeed capable of positive phototaxis with a mean vector (± 95% confidence interval) of 352°± 2.2, where 0/360° indicates the position of the light source. A study of three strains (UTEX 1688, CCMP 1326, and MBL07) of K. foliaceum showed that the eyespot in two of these strains has degenerated following decades in culture. Thus, previous studies have failed to report positive phototaxis due to loss of directionality caused by the degenerated eyespot. The results are discussed in a broader context and we conclude that studies on algal morphology and physiology may result in erroneous conclusions if based on algal cultures maintained under laboratory conditions for extended periods. © 2013 The Author(s) Journal of Eukaryotic Microbiology © 2013 International Society of Protistologists.

Cell death in a harmful algal bloom causing species Alexandrium tamarense upon an algicidal bacterium induction.

PubMed

Zhang, Huajun; Lv, Jinglin; Peng, Yun; Zhang, Su; An, Xinli; Xu, Hong; Zhang, Jun; Tian, Yun; Zheng, Wei; Zheng, Tianling

2014-09-01

Harmful algal blooms occur throughout the world, destroying aquatic ecosystems and threatening human health. The culture supernatant of the marine algicidal bacteria DHQ25 was able to lysis dinoflagellate Alexandrium tamarense. Loss of photosynthetic pigments, accompanied by a decline in Photosystem II (PSII) photochemical efficiency (Fv/Fm), in A. tamarense was detected under bacterial supernatant stress. Transmission electron microscope analysis showed obvious morphological modifications of chloroplast dismantling as a part of the algicidal process. The PSII electron transport chain was seriously blocked, with its reaction center damaged. This damage was detected in a relative transcriptional level of psbA and psbD genes, which encode the D1 and D2 proteins in the PSII reaction center. And the block in the electron transport chain of PSII might generate excessive reactive oxygen species (ROS) which could destroy the membrane system and pigment synthesis and activated enzymic antioxidant systems including superoxide dismutase (SOD) and catalase (CAT). This study indicated that marine bacteria with indirect algicidal activity played an important role in the changes of photosynthetic process in a harmful algal bloom species.

Influence of the Cell Wall on Intracellular Delivery to Algal Cells by Electroporation and Sonication

PubMed Central

Azencott, Harold R.; Peter, Gary F.; Prausnitz, Mark R.

2007-01-01

To assess the cell wall’s role as a barrier to intracellular delivery, wild-type Chlamydomonas reinhardtii algal cells and mutant cells lacking a cell wall were exposed to electroporation or sonication. Flow cytometry determined intracellular uptake of calcein and bovine serum albumin (BSA) and loss of cell viability as functions of electroporation transmembrane potential and acoustic energy. Electroporation of wild-type cells increased calcein uptake with increasing transmembrane potential, but delivered much less BSA. Electroporation of wall-deficient cells had similar effects on calcein uptake, but increased BSA uptake as much as 7.5-fold relative to wild-type cells, which indicated that the cell wall was a significant barrier to BSA delivery during electroporation. Sonication of wild-type cells caused calcein and BSA uptake at similar levels. This suggests that the cell wall barrier to BSA delivery can be overcome by sonication. Increased electroporation transmembrane potential or acoustic energy also caused increased loss of cell viability, where wall-deficient cells were especially susceptible to lysis. Overall, we believe this is the first study to compare the effects of electroporation and sonication in a direct fashion in any cell type. Specifically, these findings suggest that electroporation primarily transports molecules across the plasma membrane, because its mechanism is specific to lipid bilayer disruption, whereas sonication transports molecules across both the plasma membrane and cell wall, because it non-specifically disrupts cell-surface barriers. PMID:17602827

Harmful Algal Blooms

USGS Publications Warehouse

Graham, Jennifer L.

2007-01-01

What are Harmful Algal Blooms (HABs)? Freshwater and marine harmful algal blooms (HABs) can occur anytime water use is impaired due to excessive accumulations of algae. HAB occurrence is affected by a complex set of physical, chemical, biological, hydrological, and meteorological conditions making it difficult to isolate specific causative environmental factors. Potential impairments include reduction in water quality, accumulation of malodorous scums in beach areas, algal production of toxins potent enough to poison both aquatic and terrestrial organisms, and algal production of taste-and-odor compounds that cause unpalatable drinking water and fish. HABs are a global problem, and toxic freshwater and (or) marine algae have been implicated in human and animal illness and death in over 45 countries worldwide and in at least 27 U.S. States (Yoo and others, 1995; Chorus and Bartram, 1999; Huisman and others, 2005).

Algal Data from Selected Sites in the Upper Colorado River Basin, Colorado, Water Years 1996-97

USGS Publications Warehouse

Mize, Scott V.; Deacon, Jeffrey R.

2001-01-01

Algal community samples were collected at 15 sites in the Upper Colorado River Basin in Colorado as part of the National Water-Quality Assessment Program during water years 1996-97. Sites sampled were located in two physiographic provinces, the Southern Rocky Mountains and the Colorado Plateaus, and represented agricultural, mining, urban, and mixed land uses and background conditions. Algal samples were collected once per year during low-flow conditions. Quantitative algal samples were collected within two targeted instream habitat types including a taxonomically richest-targeted habitat and a depositional-targeted habitat. This report presents the algal community data collected at the fixed sites in the Upper Colorado River Basin study unit. Algal data include densities (abundance of cells per square centimeter of substrate) and biovolumes (cubic micrometers of cells per square centimeter of substrate) for the two habitat types. Quality-assurance and quality-control results for algal samples indicate that the largest sampling variability tends to occur in samples from small streams.

Parasites in algae mass culture

PubMed Central

Carney, Laura T.; Lane, Todd W.

2014-01-01

Parasites are now known to be ubiquitous across biological systems and can play an important role in modulating algal populations. However, there is a lack of extensive information on their role in artificial ecosystems such as algal production ponds and photobioreactors. Parasites have been implicated in the demise of algal blooms. Because individual mass culture systems often tend to be unialgal and a select few algal species are in wide scale application, there is an increased potential for parasites to have a devastating effect on commercial scale monoculture. As commercial algal production continues to expand with a widening variety of applications, including biofuel, food and pharmaceuticals, the parasites associated with algae will become of greater interest and potential economic impact. A number of important algal parasites have been identified in algal mass culture systems in the last few years and this number is sure to grow as the number of commercial algae ventures increases. Here, we review the research that has identified and characterized parasites infecting mass cultivated algae, the techniques being proposed and or developed to control them, and the potential impact of parasites on the future of the algal biomass industry. PMID:24936200

Climate Change and Algal Blooms =

NASA Astrophysics Data System (ADS)

Lin, Shengpan

Algal blooms are new emerging hazards that have had important social impacts in recent years. However, it was not very clear whether future climate change causing warming waters and stronger storm events would exacerbate the algal bloom problem. The goal of this dissertation was to evaluate the sensitivity of algal biomass to climate change in the continental United States. Long-term large-scale observations of algal biomass in inland lakes are challenging, but are necessary to relate climate change to algal blooms. To get observations at this scale, this dissertation applied machine-learning algorithms including boosted regression trees (BRT) in remote sensing of chlorophyll-a with Landsat TM/ETM+. The results show that the BRT algorithm improved model accuracy by 15%, compared to traditional linear regression. The remote sensing model explained 46% of the total variance of the ground-measured chlorophyll- a in the first National Lake Assessment conducted by the US Environmental Protection Agency. That accuracy was ecologically meaningful to study climate change impacts on algal blooms. Moreover, the BRT algorithm for chlorophyll- a would not have systematic bias that is introduced by sediments and colored dissolved organic matter, both of which might change concurrently with climate change and algal blooms. This dissertation shows that the existing atmospheric corrections for Landsat TM/ETM+ imagery might not be good enough to improve the remote sensing of chlorophyll-a in inland lakes. After deriving long-term algal biomass estimates from Landsat TM/ETM+, time series analysis was used to study the relations of climate change and algal biomass in four Missouri reservoirs. The results show that neither temperature nor precipitation was the only factor that controlled temporal variation of algal biomass. Different reservoirs, even different zones within the same reservoir, responded differently to temperature and precipitation changes. These findings were further

Exploring the Utilization of Complex Algal Communities to Address Algal Pond Crash and Increase Annual Biomass Production for Algal Biofuels

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

Hamilton, Cyd E.

2014-03-25

This white paper briefly reviews the research literature exploring complex algal communities as a means of increasing algal biomass production via increased tolerance, resilience, and resistance to a variety of abiotic and biotic perturbations occurring within harvesting timescales. This paper identifies what data are available and whether more research utilizing complex communities is needed to explore the potential of complex algal community stability (CACS) approach as a plausible means to increase biomass yields regardless of ecological context and resulting in decreased algal-based fuel prices by reducing operations costs. By reviewing the literature for what we do and do not know,more » in terms of CACS methodologies, this report will provide guidance for future research addressing pond crash phenomena.« less

Mass algal culture system

DOEpatents

Raymond, Lawrence P.

1982-01-01

An apparatus and process for the culture of algae in a liquid medium is disclosed. The medium circulates through an open trough and is exposed to an atmosphere which is temperature regulated. The nutrient content of the liquid medium is regulated to control the chemical composition growth and reproduction characteristics of the cultured algae. Before it is allowed to strike the medium, sunlight is passed through a filter to remove wavelengths which are not photosynthetically active. Heat energy can be recovered from the filter.

Mass algal culture system

DOEpatents

Raymond, Lawrence P.

1981-01-01

An apparatus and process for the culture of algae in a liquid medium is disclosed. The medium circulates through an open trough and is exposed to an atmosphere which is temperature regulated. The nutrient content of the liquid medium is regulated to control the chemical composition growth and reproduction characteristics of the cultured algae. Before it is allowed to strike the medium, sunlight is passed through a filter to remove wavelengths which are not photosynthetically active. Heat energy can be recovered from the filter.

Natural organic matter differently modulates growth of two closely related coccal green algal species.

PubMed

Karasyova, Tatyana A; Klose, Edgar O; Menzel, Ralph; Steinberg, Christian E W

2007-03-01

Humic substances (HS) comprise the majority of dead and living organic carbon, including organisms. In the environment, they are considered to be chemically inert or at least refractory. Recent papers, however, show that HS (including natural organic matter-NOM, isolated by reverse osmosis) are natural chemicals which interact with aquatic organisms. They are taken up and cause a variety of stress defense reactions which are well known from man-made chemicals. These reactions include chaperon activation, induction and modulation of biotrans-formation enzymes, or induction of antioxidant defense enzymes. One specific reaction with freshwater plants is the reduction of photosynthetic oxygen release. In this contribution, we compare the susceptibilities (cell yield) of two closely related coccal green algae, Monoraphidium convolutum and M. minutum, towards various NOM isolates. Cultures of M. convolutum and M. minutum were obtained from the algal collection of the Leibniz Institute of Freshwater Ecology and Inland Fisheries, Berlin, and from the Culture Collection of Algae, Göttingen, and maintained in a common medium. The cultures were non-axenic. The algae were exposed to 5 mg L(-1) DOC of each humic material, an environmentally realistic concentration. Cell numbers were counted microscopically in Neugebauer cuvettes in 5 replicates on days 1, 4, 7, 10, 14, and 21. Almost all NOM isolates modulated the growth of the algae. Only the NOM of a Norwegian raised peat bog lake did not reveal any significant effect with M. convolutuim. In general, the results with two algal species are by no means uniform. For instance, Suwannee River NOM causes a decrease in cell density with M. minutum, but temporarily stimulates the growth of M. convolutum. The opposite applies to Aurevann NOM: Growth increase in M. minutum, but a bi-phasic response in M. convolutum. Different responses of both Monoraphidium species must be attributed to intrinsic factors of the algae rather than only

Impact of Microalgae-Bacteria Interactions on the Production of Algal Biomass and Associated Compounds

PubMed Central

Fuentes, Juan Luis; Garbayo, Inés; Cuaresma, María; Montero, Zaida; González-del-Valle, Manuel; Vílchez, Carlos

2016-01-01

A greater insight on the control of the interactions between microalgae and other microorganisms, particularly bacteria, should be useful for enhancing the efficiency of microalgal biomass production and associated valuable compounds. Little attention has been paid to the controlled utilization of microalgae-bacteria consortia. However, the studies of microalgal-bacterial interactions have revealed a significant impact of the mutualistic or parasitic relationships on algal growth. The algal growth, for instance, has been shown to be enhanced by growth promoting factors produced by bacteria, such as indole-3-acetic acid. Vitamin B12 produced by bacteria in algal cultures and bacterial siderophores are also known to be involved in promoting faster microalgal growth. More interestingly, enhancement in the intracellular levels of carbohydrates, lipids and pigments of microalgae coupled with algal growth stimulation has also been reported. In this sense, massive algal production might occur in the presence of bacteria, and microalgae-bacteria interactions can be beneficial to the massive production of microalgae and algal products. This manuscript reviews the recent knowledge on the impact of the microalgae-bacteria interactions on the production of microalgae and accumulation of valuable compounds, with an emphasis on algal species having application in aquaculture. PMID:27213407

National Algal Biofuels Technology Roadmap

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

Ferrell, John; Sarisky-Reed, Valerie

The framework for National Algal Biofuels Technology Roadmap was constructed at the Algal Biofuels Technology Roadmap Workshop, held December 9-10, 2008, at the University of Maryland-College Park. The Workshop was organized by the Biomass Program to discuss and identify the critical challenges currently hindering the development of a domestic, commercial-scale algal biofuels industry. This Roadmap presents information from a scientific, economic, and policy perspectives that can support and guide RD&D investment in algal biofuels. While addressing the potential economic and environmental benefits of using algal biomass for the production of liquid transportation fuels, the Roadmap describes the current status ofmore » algae RD&D. In doing so, it lays the groundwork for identifying challenges that likely need to be overcome for algal biomass to be used in the production of economically viable biofuels.« less

Cultivation of algal biofilm using different lignocellulosic materials as carriers.

PubMed

Zhang, Qi; Liu, Cuixia; Li, Yubiao; Yu, Zhigang; Chen, Zhihua; Ye, Ting; Wang, Xun; Hu, Zhiquan; Liu, Shiming; Xiao, Bo; Jin, Shiping

2017-01-01

Algal biofilm technology is recently supposed to be a promising method to produce algal biomass as the feedstock for the production of biofuels. However, the carrier materials currently used to form algal biofilm are either difficult to be obtained at a low price or undurable. Commercialization of the biofilm technology for algal biomass production extremely requires new and inexpensive materials as biofilm carriers with high biomass production performances. Four types of lignocellulosic materials were investigated to evaluate their performance of acting as carriers for algal cells attachment and the relevant effects on the algal biomass production in this study. The cultivation of algal biofilm was processed in a self-designed flat plate photo-bioreactor. The biofilm production and chemical composition of the harvested biomass were determined. The surface physics properties of the materials were examined through a confocal laser-scanning microscopy. Algal biomass production varied significantly with the variation of the carriers ( P  < 0.05). All the lignocellulosic materials showed better performances in biofilm production than poly methyl methacrylate, and the application of pine sawdust as the carrier could gain the maximum biofilm productivity of 10.92 g m -2  day -1 after 16-day cultivation. In addition, 20.10-23.20% total lipid, 30.35-36.73% crude proteins, and 20.29-25.93% carbohydrate were achieved from the harvested biomasses. Biomass productivity increased linearly as the increase of surface roughness, and Wenzel's roughness factor of the tested materials, and surface roughness might significantly affect the biomass production through the size of surface morphology and the area of surface ( P  < 0.05). The results showed that lignocellulosic materials can be efficient carriers for low-cost cultivation of algal biofilm and the enhancement of biomass productivity.

Selective Algicidal Action of Peptides against Harmful Algal Bloom Species

PubMed Central

Park, Seong-Cheol; Lee, Jong-Kook; Kim, Si Wouk; Park, Yoonkyung

2011-01-01

Recently, harmful algal bloom (HAB), also termed “red tide”, has been recognized as a serious problem in marine environments according to climate changes worldwide. Many novel materials or methods to prevent HAB have not yet been employed except for clay dispersion, in which can the resulting sedimentation on the seafloor can also cause alteration in marine ecology or secondary environmental pollution. In the current study, we investigated that antimicrobial peptide have a potential in controlling HAB without cytotoxicity to harmless marine organisms. Here, antimicrobial peptides are proposed as new algicidal compounds in combating HAB cells. HPA3 and HPA3NT3 peptides which exert potent antimicrobial activity via pore forming action in plasma membrane showed that HPA3NT3 reduced the motility of algal cells, disrupted their plasma membrane, and induced the efflux of intracellular components. Against raphidoflagellate such as Heterosigma akashiwo, Chattonella sp., and C. marina, it displayed a rapid lysing action in cell membranes at 1∼4 µM within 2 min. Comparatively, its lysing effects occurred at 8 µM within 1 h in dinoflagellate such as Cochlodium polykrikoides, Prorocentrum micans, and P. minimum. Moreover, its lysing action induced the lysis of chloroplasts and loss of chlorophyll a. In the contrary, this peptide was not effective against Skeletonema costatum, harmless algal cell, even at 256 µM, moreover, it killed only H. akashiwo or C. marina in co-cultivation with S. costatum, indicating to its selective algicidal activity between harmful and harmless algal cells. The peptide was non-hemolytic against red blood cells of Sebastes schlegeli, the black rockfish, at 120 µM. HAB cells were quickly and selectively lysed following treatment of antimicrobial peptides without cytotoxicity to harmless marine organisms. Thus, the antibiotic peptides examined in our study appear to have much potential in effectively controlling HAB with minimal impact on

Biotic interactions as drivers of algal origin and evolution.

PubMed

Brodie, Juliet; Ball, Steven G; Bouget, François-Yves; Chan, Cheong Xin; De Clerck, Olivier; Cock, J Mark; Gachon, Claire; Grossman, Arthur R; Mock, Thomas; Raven, John A; Saha, Mahasweta; Smith, Alison G; Vardi, Assaf; Yoon, Hwan Su; Bhattacharya, Debashish

2017-11-01

Contents 670 I. 671 II. 671 III. 676 IV. 678 678 References 678 SUMMARY: Biotic interactions underlie life's diversity and are the lynchpin to understanding its complexity and resilience within an ecological niche. Algal biologists have embraced this paradigm, and studies building on the explosive growth in omics and cell biology methods have facilitated the in-depth analysis of nonmodel organisms and communities from a variety of ecosystems. In turn, these advances have enabled a major revision of our understanding of the origin and evolution of photosynthesis in eukaryotes, bacterial-algal interactions, control of massive algal blooms in the ocean, and the maintenance and degradation of coral reefs. Here, we review some of the most exciting developments in the field of algal biotic interactions and identify challenges for scientists in the coming years. We foresee the development of an algal knowledgebase that integrates ecosystem-wide omics data and the development of molecular tools/resources to perform functional analyses of individuals in isolation and in populations. These assets will allow us to move beyond mechanistic studies of a single species towards understanding the interactions amongst algae and other organisms in both the laboratory and the field. © 2017 The Authors. New Phytologist © 2017 New Phytologist Trust.

Recent progress and future challenges in algal biofuel production

PubMed Central

Shurin, Jonathan B.; Burkart, Michael D.; Mayfield, Stephen P.

2016-01-01

Modern society is fueled by fossil energy produced millions of years ago by photosynthetic organisms. Cultivating contemporary photosynthetic producers to generate energy and capture carbon from the atmosphere is one potential approach to sustaining society without disrupting the climate. Algae, photosynthetic aquatic microorganisms, are the fastest growing primary producers in the world and can therefore produce more energy with less land, water, and nutrients than terrestrial plant crops. We review recent progress and challenges in developing bioenergy technology based on algae. A variety of high-value products in addition to biofuels can be harvested from algal biomass, and these may be key to developing algal biotechnology and realizing the commercial potential of these organisms. Aspects of algal biology that differentiate them from plants demand an integrative approach based on genetics, cell biology, ecology, and evolution. We call for a systems approach to research on algal biotechnology rooted in understanding their biology, from the level of genes to ecosystems, and integrating perspectives from physical, chemical, and social sciences to solve one of the most critical outstanding technological problems. PMID:27781084

Harmful Algal Bloom Webinar

EPA Pesticide Factsheets

The problem is complex. Excessive nitrogen and phosphorous levels can cause harmful algal blooms. Different algal/cyanobacteria strains bloom under different conditions. Different strains produce different toxins at varying amounts.

In situ Transesterification of Microalgal Oil to Produce Algal Biodiesel

DOT National Transportation Integrated Search

2012-06-01

This research was to process whole microalgae cells for biodiesel production without first extracting lipids. The ultimate : goal is develop a novel process for algal biodiesel production directly from microalgae cells in a single step, i.e., in situ...

Algal Attributes: An Autecological Classification of Algal Taxa Collected by the National Water-Quality Assessment Program

USGS Publications Warehouse

Porter, Stephen D.

2008-01-01

Algae are excellent indicators of water-quality conditions, notably nutrient and organic enrichment, and also are indicators of major ion, dissolved oxygen, and pH concentrations and stream microhabitat conditions. The autecology, or physiological optima and tolerance, of algal species for various water-quality contaminants and conditions is relatively well understood for certain groups of freshwater algae, notably diatoms. However, applications of autecological information for water-quality assessments have been limited because of challenges associated with compiling autecological literature from disparate sources, tracking name changes for a large number of algal species, and creating an autecological data base from which algal-indicator metrics can be calculated. A comprehensive summary of algal autecological attributes for North American streams and rivers does not exist. This report describes a large, digital data file containing 28,182 records for 5,939 algal taxa, generally species or variety, collected by the U.S. Geological Survey?s National Water-Quality Assessment (NAWQA) Program. The data file includes 37 algal attributes classified by over 100 algal-indicator codes or metrics that can be calculated easily with readily available software. Algal attributes include qualitative classifications based on European and North American autecological literature, and semi-quantitative, weighted-average regression approaches for estimating optima using regional and national NAWQA data. Applications of algal metrics in water-quality assessments are discussed and national quartile distributions of metric scores are shown for selected indicator metrics.

[Algal oligosaccharides ameliorate osteoporosis via up-regulation of parathyroid hormone 1-84 and vascular endothelial growth factor].

PubMed

Wang, Li; Wang, Haiya; Fang, Ningyuan

2016-06-01

To determine whether algal oligosac- charide~ affects the levels of parathyroid hormone 1-84 (PTH1-84) and vascular endothelial growth fac- tor (VEGF). An osteoporosis rat model was estab- lished via bilateral ovariectomy. The model rats were fed algal oligosaccharides (molecular weights: 600-1, 200 Da) for 4 months. Bone mineral density (BMD) was then measured. MG-63 human osteo- blastic cells were treated with algal oligosaccha- rides. The expression of PTH1-84 and VEGF was then examined. Oligosaccharide-treated cells were transfected with PTH1-84 short hairpin RNA (shR- NA), VEGF shRNA, and PTH1-84-VEGF small interfer- ing RNA (siRNA). The growth rates were then com- pared between transfected and non-transfected Algal oligosaccharides increased the BMD of the osteoporosis rat model compared with untreated controls (P < 0.05). When MG-63 cells were treated with algal oligosaccharides, the growth rate increased by 25% compared with the control group at day 3 (P < 0.05). In addition, the ex- pression of P.TH84 and VEGF was. enhanced. Con- versey w hen tecells were tranfected with PTH84 shRNA, VEGF shRNA, or PTH1-84-VEGF siR- NA, the growth rate was decreased by 17%, 35% and 70%, respectively, compared with controls at day 3 (P < 0.05). Algal oligosaccharides ameliorate osteoporosis via up-regulation of PTH1-84 and VEGF. Algal oligosaccharides should be developed as a potential drug for osteoporosis treatment.

Progress on lipid extraction from wet algal biomass for biodiesel production.

PubMed

Ghasemi Naghdi, Forough; González González, Lina M; Chan, William; Schenk, Peer M

2016-11-01

Lipid recovery and purification from microalgal cells continues to be a significant bottleneck in biodiesel production due to high costs involved and a high energy demand. Therefore, there is a considerable necessity to develop an extraction method which meets the essential requirements of being safe, cost-effective, robust, efficient, selective, environmentally friendly, feasible for large-scale production and free of product contamination. The use of wet concentrated algal biomass as a feedstock for oil extraction is especially desirable as it would avoid the requirement for further concentration and/or drying. This would save considerable costs and circumvent at least two lengthy processes during algae-based oil production. This article provides an overview on recent progress that has been made on the extraction of lipids from wet algal biomass. The biggest contributing factors appear to be the composition of algal cell walls, pre-treatments of biomass and the use of solvents (e.g. a solvent mixture or solvent-free lipid extraction). We compare recently developed wet extraction processes for oleaginous microalgae and make recommendations towards future research to improve lipid extraction from wet algal biomass. © 2016 The Authors. Microbial Biotechnology published by John Wiley & Sons Ltd and Society for Applied Microbiology.

Prechlorination of algae-laden water: The effects of transportation time on cell integrity, algal organic matter release, and chlorinated disinfection byproduct formation.

PubMed

Qi, Jing; Lan, Huachun; Liu, Ruiping; Miao, Shiyu; Liu, Huijuan; Qu, Jiuhui

2016-10-01

The prechlorination-induced algal organic matter (AOM) released from Microcystis aeruginosa (M. aeruginosa) cells has been reported to serve as a source of precursors for chlorinated disinfection byproducts (DBPs). However, previous studies have mainly focused on the precursors either extracted directly from the cell suspension or derived immediately after algal suspension prechlorination. This study aims to investigate the impacts of water transportation time after algal suspension prechlorination on cell integrity, AOM release, and DBP formation during the dissolved phase chlorination. The damage to cell integrity after prechlorination was indicated to depend not only on chlorine dose but also on transportation time. The highest dissolved organic carbon (DOC) and dissolved organic nitrogen (DON) values were observed at 2 mg/L chlorine preoxidation before transportation, but were obtained at 0.4 mg/L chlorine after 480-min simulated transportation. The variation of DON with transportation time was indicated to be mainly influenced by the small molecular weight nitrogenous organic compounds, such as amino acids. Additionally, formation of the corresponding chlorinated carbonaceous disinfection byproducts (C-DBPs) and nitrogenous disinfection byproducts (N-DBPs) during the dissolved phase chlorination showed the same variation tendency as DOC and DON respectively. The highest C-DBP (98.4 μg/L) and N-DBP (5.5 μg/L) values were obtained at 0.4 mg/L chlorine preoxidation after 480-min simulated transportation. Therefore, when prechlorination is applied for algae-laden water pretreatment, not only chlorine dose but also transportation time needs to be considered with regard to their effects on cell integrity, AOM release, and chlorinated DBP formation. Copyright © 2016. Published by Elsevier Ltd.

Cell culture contamination.

PubMed

Stacey, Glyn N

2011-01-01

Microbial contamination is a major issue in cell culture, but there are a range of procedures which can be adopted to prevent or eliminate contamination. Contamination may arise from the operator and the laboratory environment, from other cells used in the laboratory, and from reagents. Some infections may present a risk to laboratory workers: containment and aseptic technique are the key defence against such risks. Remedial management of suspected infection may simply mean discarding a single potentially infected culture. However, if a more widespread problem is identified, then all contaminated cultures and associated unused media that have been opened during this period should be discarded, equipment should be inspected and cleaned, cell culture operations reviewed, and isolation from other laboratories instituted until the problem is solved. Attention to training of staff, laboratory layout, appropriate use of quarantine for new cultures or cell lines, cleaning and maintenance, and quality control are important factors in preventing contamination in cell culture laboratories.

Monitoring and removal of cyanobacterial toxins from drinking water by algal-activated carbon.

PubMed

Ibrahim, Wael M; Salim, Emad H; Azab, Yahia A; Ismail, Abdel-Hamid M

2016-10-01

Microcystins (MCs) are the most potent toxins that can be produced by cyanobacteria in drinking water supplies. This study investigated the abundance of toxin-producing algae in 11 drinking water treatment plants (DWTPs). A total of 26 different algal taxa were identified in treated water, from which 12% were blue green, 29% were green, and 59% were diatoms. MC levels maintained strong positive correlations with number of cyanophycean cells in raw and treated water of different DWTPs. Furthermore, the efficiency of various algal-based adsorbent columns used for the removal of these toxins was evaluated. The MCs was adsorbed in the following order: mixed algal-activated carbon (AAC) ≥ individual AAC > mixed algal powder > individual algal powder. The results showed that the AAC had the highest efficient columns capable of removing 100% dissolved MCs from drinking water samples, thereby offering an economically feasible technology for efficient removal and recovery of MCs in DWTPs. © The Author(s) 2015.

The role of dissolved organic matter (DOM) quality in the growth enhancement of Alexandrium fundyense (Dinophyceae) in laboratory culture(1).

PubMed

Cawley, Kaelin M; Koerfer, Verena; McKnight, Diane M

2013-06-01

Several algal species responsible for harmful algal blooms (HABs), such as Alexandrium fundyense, are mixotrophic under certain environmental conditions. The ability to switch between photosynthetic and heterotrophic modes of growth may play a role in the development of HABs in coastal regions. We examined the influence of humic dissolved organic matter (HDOM) derived from terrestrial (plant/soil) and microbial sources on the growth of A. fundyense. We found that a terrestrially derived HDOM, Suwannee River humic acid (SRHA), did enhance A. fundyense growth; however, a microbially derived HDOM, Pony Lake fulvic acid (PLFA) did not enhance growth. A. fundyense grows in association with bacteria in culture and we observed that bacterial cell densities were much lower in A. fundyense cultures than in bacteria-only cultures, consistent with bacterial grazing by A. fundyense in culture. In bacteria-only cultures with added algal exudates (EX), the addition of PLFA and SRHA resulted in a slight increase in bacterial cell density compared to cultures without HDOM added. Changes over time in the chemical quality of the HDOM in the A. fundyense cultures reflected contributions of microbially derived material with similar characteristics as the PLFA. Overall, these results suggest that the chemical differences between SRHA and PLFA are responsible for the greater effect of SRHA on A. fundyense growth, and that the differential effect is not a result of an effect on the growth of associated bacteria. © 2013 Phycological Society of America.

Introduction to cell culture.

PubMed

Philippeos, Christina; Hughes, Robin D; Dhawan, Anil; Mitry, Ragai R

2012-01-01

The basics of cell culture as applied to human cells are discussed. Biosafety when working with human tissue, which is often pathogenic, is important. The requirements for a tissue culture laboratory are described, particularly the range of equipment needed to carry out cell isolation, purification, and culture. Steps must be taken to maintain aseptic conditions to prevent contamination of cultures with micro-organisms. Basic cell-handling techniques are discussed, including choice of media, primary culture, and cryopreservation of cells so they can be stored for future use. Common assays which are used to determine cell viability and activity are considered.

Coral host cells acidify symbiotic algal microenvironment to promote photosynthesis

PubMed Central

Barott, Katie L.; Venn, Alexander A.; Perez, Sidney O.; Tambutté, Sylvie; Tresguerres, Martin

2015-01-01

Symbiotic dinoflagellate algae residing inside coral tissues supply the host with the majority of their energy requirements through the translocation of photosynthetically fixed carbon. The algae, in turn, rely on the host for the supply of inorganic carbon. Carbon must be concentrated as CO2 in order for photosynthesis to proceed, and here we show that the coral host plays an active role in this process. The host-derived symbiosome membrane surrounding the algae abundantly expresses vacuolar H+-ATPase (VHA), which acidifies the symbiosome space down to pH ∼4. Inhibition of VHA results in a significant decrease in average H+ activity in the symbiosome of up to 75% and a significant reduction in O2 production rate, a measure of photosynthetic activity. These results suggest that host VHA is part of a previously unidentified carbon concentrating mechanism for algal photosynthesis and provide mechanistic evidence that coral host cells can actively modulate the physiology of their symbionts. PMID:25548188

Advanced Algal Systems Fact Sheet

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

None

2016-06-01

Research and development (R&D) on advanced algal biofuels and bioproducts presents an opportunity to sustainably expand biomass resource potential in the United States. The Bioenergy Technologies Office’s (BETO’s) Advanced Algal Systems Program is carrying out a long-term, applied R&D strategy to lower the costs of algal biofuel production by working with partners to develop revolutionary technologies and conduct crosscutting analyses to better understand the potential

Recycling algae to improve species control and harvest efficiency from a high rate algal pond.

PubMed

Park, J B K; Craggs, R J; Shilton, A N

2011-12-15

This paper investigates the influence of recycling gravity harvested algae on species dominance and harvest efficiency in wastewater treatment High Rate Algal Ponds (HRAP). Two identical pilot-scale HRAPs were operated over one year either with (HRAP(r)) or without (HRAP(c)) harvested algal biomass recycling. Algae were harvested from the HRAP effluent in algal settling cones (ASCs) and harvest efficiency was compared to settlability in Imhoff cones five times a week. A microscopic image analysis technique was developed to determine relative algal dominance based on biovolume and was conducted once a month. Recycling of harvested algal biomass back to the HRAP(r) maintained the dominance of a single readily settleable algal species (Pediastrum sp.) at >90% over one year (compared to the control with only 53%). Increased dominance of Pediastrum sp. greatly improved the efficiency of algal harvest (annual average of >85% harvest for the HRAP(r) compared with ∼60% for the control). Imhoff cone experiments demonstrated that algal settleability was influenced by both the dominance of Pediastrum sp. and the species composition of remaining algae. Algal biomass recycling increased the average size of Pediastrum sp. colonies by 13-30% by increasing mean cell residence time. These results indicate that recycling gravity harvested algae could be a simple and effective operational strategy to maintain the dominance of readily settleable algal species, and enhance algal harvest by gravity sedimentation. Copyright © 2011 Elsevier Ltd. All rights reserved.

Study of cnidarian-algal symbiosis in the "omics" age.

PubMed

Meyer, Eli; Weis, Virginia M

2012-08-01

The symbiotic associations between cnidarians and dinoflagellate algae (Symbiodinium) support productive and diverse ecosystems in coral reefs. Many aspects of this association, including the mechanistic basis of host-symbiont recognition and metabolic interaction, remain poorly understood. The first completed genome sequence for a symbiotic anthozoan is now available (the coral Acropora digitifera), and extensive expressed sequence tag resources are available for a variety of other symbiotic corals and anemones. These resources make it possible to profile gene expression, protein abundance, and protein localization associated with the symbiotic state. Here we review the history of "omics" studies of cnidarian-algal symbiosis and the current availability of sequence resources for corals and anemones, identifying genes putatively involved in symbiosis across 10 anthozoan species. The public availability of candidate symbiosis-associated genes leaves the field of cnidarian-algal symbiosis poised for in-depth comparative studies of sequence diversity and gene expression and for targeted functional studies of genes associated with symbiosis. Reviewing the progress to date suggests directions for future investigations of cnidarian-algal symbiosis that include (i) sequencing of Symbiodinium, (ii) proteomic analysis of the symbiosome membrane complex, (iii) glycomic analysis of Symbiodinium cell surfaces, and (iv) expression profiling of the gastrodermal cells hosting Symbiodinium.

Ultrasound pretreatment of filamentous algal biomass for enhanced biogas production.

PubMed

Lee, Kwanyong; Chantrasakdakul, Phrompol; Kim, Daegi; Kong, Mingeun; Park, Ki Young

2014-06-01

The filamentous alga Hydrodictyon reticulatum harvested from a bench-scale wastewater treatment pond was used to evaluate biogas production after ultrasound pretreatment. The effects of ultrasound pretreatment at a range of 10-5000 J/mL were tested with harvested H. reticulatum. Cell disruption by ultrasound was successful and showed a higher degree of disintegration at a higher applied energy. The range of 10-5000 J/mL ultrasound was able to disintegrated H. reticulatum and the soluble COD was increased from 250 mg/L to 1000 mg/L at 2500 J/mL. The disintegrated algal biomass was digested for biogas production in batch experiments. Both cumulative gas generation and volatile solids reduction data were obtained during the digestion. Cell disintegration due to ultrasound pretreatment increased the specific biogas production and degradation rates. Using the ultrasound approach, the specific methane production at a dose of 40 J/mL increased up to 384 mL/g-VS fed that was 2.3 times higher than the untreated sample. For disintegrated samples, the volatile solids reduction was greater with increased energy input, and the degradation increased slightly to 67% at a dose of 50 J/mL. The results also indicate that disintegration of the algal cells is the essential step for efficient anaerobic digestion of algal biomass. Copyright © 2013 Elsevier Ltd. All rights reserved.

Amino acids in cell wall of Gram-positive bacterium Micrococcus sp. hsn08 with flocculation activity on Chlorella vulgaris biomass.

PubMed

Li, Yi; Xu, Yanting; Zheng, Tianling; Wang, Hailei

2018-02-01

The aim of this work was to investigate the flocculation mechanism by Gram-positive bacterium, Micrococcus sp. hsn08 as a means for harvesting Chlorella vulgaris biomass. Bacterial cells of Micrococcus sp. hsn08 were added into algal culture to harvest algal cells through direct contacting with algae to form flocs. Viability dependence test confirmed that flocculation activity does not depend on live bacteria, but on part of the peptidoglycan. The further investigation has determined that amino acids in cell wall play an important role to flocculate algal cells. Positively charged calcium can combine bacterial and algal cells together, and form a bridge between them, thereby forming the flocs, suggesting that ions bridging is the main flocculation mechanism. These results suggest that bacterial cells of Micrococcus sp. hsn08 can be applied to harvest microalgae biomass with the help of amino acids in cell wall. Copyright © 2017 Elsevier Ltd. All rights reserved.

Increasing cell culture population doublings for long-term growth of finite life span human cell cultures

DOEpatents

Stampfer, Martha R.; Garbe, James C.

2016-06-28

Cell culture media formulations for culturing human epithelial cells are herein described. Also described are methods of increasing population doublings in a cell culture of finite life span human epithelial cells and prolonging the life span of human cell cultures. Using the cell culture media disclosed alone and in combination with addition to the cell culture of a compound associated with anti-stress activity achieves extended growth of pre-stasis cells and increased population doublings and life span in human epithelial cell cultures.

Increasing cell culture population doublings for long-term growth of finite life span human cell cultures

DOEpatents

Stampfer, Martha R; Garbe, James C

2015-02-24

Cell culture media formulations for culturing human epithelial cells are herein described. Also described are methods of increasing population doublings in a cell culture of finite life span human epithelial cells and prolonging the life span of human cell cultures. Using the cell culture media disclosed alone and in combination with addition to the cell culture of a compound associated with anti-stress activity achieves extended growth of pre-stasis cells and increased population doublings and life span in human epithelial cell cultures.

Atmosphere stabilization and element recycle in an experimental mouse-algal system

NASA Technical Reports Server (NTRS)

Smernoff, David T.

1986-01-01

Life support systems based on bioregeneration rely on the control and manipulation of organisms. Experiments conducted with a gas-closed mouse-algal system designed to investigate principles of photosynthetic gas exchange focus primarily on observing gas exchange phenomena under varying algal environmental conditions and secondarily on studying element cycling through compartments of the experimental system. Inherent instabilities exit between the uptake and release of carbon dioxide CO2 and oxygen O2 by the mouse and algae. Variations in light intensity and cell density alter the photosynthetic rate of the algae and enable maintenance of physiologic concentrations of CO2 and O2. Different nitrogen sources (urea and nitrate) result in different algal assimilatory quotients (AQ). Combinations of photosynthetic rate and AQ ratio manipulations have been examined for their potential in stabilizing atmospheric gas concentrations in the gas-closed algal-mouse system. Elemental mass balances through the experimental systems compartments are being studied with the concurrent development of a mathematical simulation model. Element cycling experiments include quantification of elemental flows through system compartments and wet oxidation of system waste materials for use as an algal nutrient source. Oxidized waste products demonstrate inhibitory properties although dilution has been shown to allow normal growth.

Significance of different carbon forms and carbonic anhydrase activity in monitoring and prediction of algal blooms in the urban section of Jialing River, Chongqing, China.

PubMed

Nie, Yudong; Zhang, Zhi; Shen, Qian; Gao, Wenjin; Li, Yingfan

2016-05-18

The Three Gorges Dam is one of the largest hydroelectric power plants worldwide; its reservoir was preliminarily impounded in 2003 and finally impounded to 175 m in 2012. The impoundment caused some environmental problems, such as algal blooms. Carbonic anhydrase (CA) is an important biocatalyst in the carbon utilization by algae and plays an important role in algal blooms. CA has received considerable attention for its role in red tides in oceans, but less investigation has been focused on its role in algal blooms in fresh water. In this study, the seasonal variation of water quality parameters, different carbon forms, carbonic anhydrase activity (CAA), and the algal cell density of four sampling sites in the urban section of the Jialing River were investigated from November 1, 2013 to October 31, 2014. Results indicated that CAA exhibited a positive correlation with dissoluble organic carbon (DOC), pH, and temperature, but a negative correlation with CO2 and dissoluble inorganic carbon (DIC). Algal cell density exhibited a positive correlation with flow velocity (V), pH, particulate organic carbon (POC), and CAA, a negative correlation with CO2, and a negative partial correlation with DIC. The relationship between CAA and algal cell density for the entire year can be described as cells = 23.278CAA - 42.666POC + 139.547pH - 1057.106. The algal bloom prediction model for the key control period can be described as cells = -45.895CAA + 776.103V- 29.523DOC + 14.219PIC + 35.060POC + 19.181 (2 weeks in advance) and cells = 69.200CAA + 203.213V + 4.184CO2 + 38.911DOC + 40.770POC - 189.567 (4 weeks in advance). The findings in this study demonstrate that the carbon utilization by algae is conducted by CA and provide a new method of monitoring algal cell density and predicting algal blooms.

Time-response of cultured deep-sea benthic foraminifera to different algal diets

NASA Astrophysics Data System (ADS)

Heinz, P.; Hemleben, Ch; Kitazato, H.

2002-03-01

The vertical distribution of benthic foraminifera in the surface sediment is influenced by environmental factors, mainly by food and oxygen supply. An experiment of three different time series was performed to investigate the response of deep-sea benthic foraminifera to simulated phytodetritus pulses under stable oxygen concentrations. Each series was fed constantly with one distinct algal species in equivalent amounts. The temporal reactions of the benthic foraminifera with regard to the vertical distribution in the sediment, the total number, and the species composition were observed and compared within the three series. Additionally, oxygen contents and bacterial cell numbers were measured to ensure that these factors were invariable and did not influence foraminiferal communities. The addition of algae leads to higher population densities 21 days after food was added. Higher numbers of individuals were probably caused by higher organic levels, which in turn induced reproduction. A stronger response is found after feeding with Amphiprora sp. and Pyramimonas sp., compared to Dunaliella tertiolecta. At a constant high oxygen supply, no migration to upper layers was observed after food addition, and more individuals were found in deeper layers. The laboratory results thus agree with the predictions of the TROX-model. An epifaunal microhabitat preference was shown for Adercotryma glomerata. Hippocrepina sp. was spread over the entire sediment depth with a shallow infaunal maximum. Melonis barleeanum preferred a deeper infaunal habitat. Bacterial cell concentrations were stable during the laboratory experiments and showed no significant response to higher organic fluxes.

Continuous energy recovery and nutrients removal from molasses wastewater by synergistic system of dark fermentation and algal culture under various fermentation types.

PubMed

Ren, Hong-Yu; Kong, Fanying; Ma, Jun; Zhao, Lei; Xie, Guo-Jun; Xing, Defeng; Guo, Wan-Qian; Liu, Bing-Feng; Ren, Nan-Qi

2018-03-01

Synergistic system of dark fermentation and algal culture was initially operated at batch mode to investigate the energy production and nutrients removal from molasses wastewater in butyrate-type, ethanol-type and propionate-type fermentations. Butyrate-type fermentation was the most appropriate fermentation type for the synergistic system and exhibited the accumulative hydrogen volume of 658.3 mL L -1 and hydrogen yield of 131.7 mL g -1 COD. By-products from dark fermentation (mainly acetate and butyrate) were further used to cultivate oleaginous microalgae. The maximum algal biomass and lipid content reached 1.01 g L -1 and 38.5%, respectively. In continuous operation, the synergistic system was stable and efficient, and energy production increased from 8.77 kJ L -1  d -1 (dark fermentation) to 17.3 kJ L -1  d -1 (synergistic system). Total COD, TN and TP removal efficiencies in the synergistic system reached 91.1%, 89.1% and 85.7%, respectively. This study shows the potential of the synergistic system in energy recovery and wastewater treatment. Copyright © 2017 Elsevier Ltd. All rights reserved.

Bacilysin from Bacillus amyloliquefaciens FZB42 Has Specific Bactericidal Activity against Harmful Algal Bloom Species

PubMed Central

Wu, Liming; Wu, Huijun; Chen, Lina; Xie, Shanshan; Zang, Haoyu; Borriss, Rainer

2014-01-01

Harmful algal blooms, caused by massive and exceptional overgrowth of microalgae and cyanobacteria, are a serious environmental problem worldwide. In the present study, we looked for Bacillus strains with sufficiently strong anticyanobacterial activity to be used as biocontrol agents. Among 24 strains, Bacillus amyloliquefaciens FZB42 showed the strongest bactericidal activity against Microcystis aeruginosa, with a kill rate of 98.78%. The synthesis of the anticyanobacterial substance did not depend on Sfp, an enzyme that catalyzes a necessary processing step in the nonribosomal synthesis of lipopeptides and polyketides, but was associated with the aro gene cluster that is involved in the synthesis of the sfp-independent antibiotic bacilysin. Disruption of bacB, the gene in the cluster responsible for synthesizing bacilysin, or supplementation with the antagonist N-acetylglucosamine abolished the inhibitory effect, but this was restored when bacilysin synthesis was complemented. Bacilysin caused apparent changes in the algal cell wall and cell organelle membranes, and this resulted in cell lysis. Meanwhile, there was downregulated expression of glmS, psbA1, mcyB, and ftsZ—genes involved in peptidoglycan synthesis, photosynthesis, microcystin synthesis, and cell division, respectively. In addition, bacilysin suppressed the growth of other harmful algal species. In summary, bacilysin produced by B. amyloliquefaciens FZB42 has anticyanobacterial activity and thus could be developed as a biocontrol agent to mitigate the effects of harmful algal blooms. PMID:25261512

Algal conditions in the Caloosahatchee River (1975-79), Lake Okeechobee to Franklin Lock, Florida

USGS Publications Warehouse

McPherson, Benjamin F.; La Rose, Henry R.

1982-01-01

Maximum numbers of suspended algae occurred in late spring and early summer, in each of the years 1975-79, in the Caloosahatchee River. Numbers exceeded 100,000 cells per milliliter at all stations sometime during the study. Concentrations decreased during late summer and autumn and were low during winter, except in January 1979 when numbers at most sites exceeded 100,000 cells per milliliter. The January 1979 bloom coincided with large discharges from Lake Okeechobee. During previous winters, discharges and algal numbers were lower. During other seasons, algal blooms occurred most frequently under low-flow or stagnant conditions. The upstream site at Moore Haven, which had the least discharge and was most stagnant, had consistently higher algal concentrations than downstream sites. Blue-green algae were dominant in the river during the summer at the upstream site throughout the year. The percentage of blue-green algae decreased downstream. Concentrations of nitrite plus nitrate nitrogen were inversely correlated with concentrations of algae and decreased to near zero during algal blooms. The low concentrations of these forms of inorganic nitrogen relative to other major nutrients probably favor blue-green algae and limit growth of other algae. Contributions by the basin tributaries to the nutritive condition of the river were small because concentrations of nutrients, algal growth potential, and algae in the tributaries were generally less than those in the river. (USGS)

Energetic potential of algal biomass from high-rate algal ponds for the production of solid biofuels.

PubMed

Costa, Taynan de Oliveira; Calijuri, Maria Lúcia; Avelar, Nayara Vilela; Carneiro, Angélica de Cássia de Oliveira; de Assis, Letícia Rodrigues

2017-08-01

In this investigation, chemical characteristics, higher, lower and net heating value, bulk and energy density, and thermogravimetric analysis were applied to study the thermal characteristics of three algal biomasses. These biomasses, grown as by-products of wastewater treatment in high-rate algal ponds (HRAPs), were: (i) biomass produced in domestic effluent and collected directly from an HRAP (PO); (ii) biomass produced in domestic effluent in a mixed pond-panel system and collected from the panels (PA); and (iii) biomass originating from the treatment effluent from the meat processing industry and collected directly from an HRAP (IN). The biomass IN was the best alternative for thermal power generation. Subsequently, a mixture of the algal biomasses and Jatropha epicarp was used to produce briquettes containing 0%, 25%, 50%, 75%, and 100% of algal biomass, and their properties were evaluated. In general, the addition of algal biomass to briquettes decreased both the hygroscopicity and fixed carbon content and increased the bulk density, ash content, and energy density. A 50% proportion of biomass IN was found to be the best raw material for producing briquettes. Therefore, the production of briquettes consisting of algal biomass and Jatropha epicarp at a laboratory scale was shown to be technically feasible.

Removal of algal blooms from freshwater by the coagulation-magnetic separation method.

PubMed

Liu, Dan; Wang, Peng; Wei, Guanran; Dong, Wenbo; Hui, Franck

2013-01-01

This research investigated the feasibility of changing waste into useful materials for water treatment and proposed a coagulation-magnetic separation technique. This technique was rapid and highly effective for clearing up harmful algal blooms in freshwater and mitigating lake eutrophication. A magnetic coagulant was synthesized by compounding acid-modified fly ash with magnetite (Fe(3)O(4)). Its removal effects on algal cells and dissolved organics in water were studied. After mixing, coagulation, and magnetic separation, the flocs obtained from the magnet surface were examined by SEM. Treated samples were withdrawn for the content determination of chlorophyll-a, turbidity, chemical oxygen demand (COD), total nitrogen, and total phosphorus. More than 99 % of algal cells were removed within 5 min after the addition of magnetic coagulant at optimal loadings (200 mg L(-1)). The removal efficiencies of COD, total nitrogen, and phosphorus were 93, 91, and 94 %, respectively. The mechanism of algal removal explored preliminarily showed that the magnetic coagulant played multiple roles in mesoporous adsorption, netting and bridging, as well as high magnetic responsiveness to a magnetic field. The magnetic-coagulation separation method can rapidly and effectively remove algae from water bodies and greatly mitigate eutrophication of freshwater using a new magnetic coagulant. The method has good performance, is low cost, can turn waste into something valuable, and provides reference and directions for future pilot and production scale-ups.

Alga-PrAS (Algal Protein Annotation Suite): A Database of Comprehensive Annotation in Algal Proteomes

PubMed Central

Kurotani, Atsushi; Yamada, Yutaka

2017-01-01

Algae are smaller organisms than land plants and offer clear advantages in research over terrestrial species in terms of rapid production, short generation time and varied commercial applications. Thus, studies investigating the practical development of effective algal production are important and will improve our understanding of both aquatic and terrestrial plants. In this study we estimated multiple physicochemical and secondary structural properties of protein sequences, the predicted presence of post-translational modification (PTM) sites, and subcellular localization using a total of 510,123 protein sequences from the proteomes of 31 algal and three plant species. Algal species were broadly selected from green and red algae, glaucophytes, oomycetes, diatoms and other microalgal groups. The results were deposited in the Algal Protein Annotation Suite database (Alga-PrAS; http://alga-pras.riken.jp/), which can be freely accessed online. PMID:28069893

Algal Accessory Pigment Detection Using AVIRIS Image-Derived Spectral Radiance Data

NASA Technical Reports Server (NTRS)

Richardson, Laurie L.; Ambrosia, Vincent G.

1996-01-01

Visual and derivative analyses of AVIRIS spectral data can be used to detect algal accessory pigments in aquatic communities. This capability extends the use of remote sensing for the study of aquatic ecosystems by allowing detection of taxonomically significant pigment signatures which yield information about the type of algae present. Such information allows remote sensing-based assessment of aquatic ecosystem health, as in the detection of nuisance blooms of cyanobacteria or toxic blooms of dinoflagellates. Remote sensing of aquatic systems has traditionally focused on quantification of chlorophyll a, a photoreactive (and light-harvesting) pigment which is common to all algae as well as cyanobacteria (bluegreen algae). Due to the ubiquitousness of this pigment within algae, chl a is routinely measured to estimate algal biomass both during ground-truthing and using various airborne or satellite based sensors, including AVIRIS. Within the remote sensing and aquatic sciences communities, ongoing research has been performed to detect algal accessory pigments for assessment of algal population composition. This research is based on the fact that many algal accessory pigments are taxonomically significant, and all are spectrally unique. Aquatic scientists have been refining pigment analysis techniques, primarily high performance liquid chromatography, or HPLC, to detect specific pigments as a time-saving alternative to individual algal cell identifications and counts. Remote sensing scientists are investigating the use of pigment signatures to construct pigment libraries analogous to mineral spectral libraries used in geological remote sensing applications. The accessory pigment approach has been used successfully in remote sensing using data from the Thematic Mapper, low-altitude, multiple channel scanners, field spectroradiometers and the AVIRIS hyperspectral scanner. Due to spectral and spatial resolution capabilities, AVIRIS is the sensor of choice for such

Plasticity of Total and Intracellular Phosphorus Quotas in Microcystis aeruginosa Cultures and Lake Erie Algal Assemblages

PubMed Central

Saxton, Matthew A.; Arnold, Robert J.; Bourbonniere, Richard A.; McKay, Robert Michael L.; Wilhelm, Steven W.

2011-01-01

Blooms of the potentially toxic cyanobacterium Microcystis are common events globally, and as a result significant resources continue to be dedicated to monitoring and controlling these events. Recent studies have shown that a significant proportion of total cell-associated phosphorus (P) in marine phytoplankton can be surface adsorbed; as a result studies completed to date do not accurately report the P demands of these organisms. In this study we measure the total cell-associated and intracellular P as well as growth rates of two toxic strains of Microcystis aeruginosa Kütz grown under a range of P concentrations. The results show that the intracellular P pool in Microcystis represents a percentage of total cell-associated P (50–90%) similar to what has been reported for actively growing algae in marine systems. Intracellular P concentrations (39–147 fg cell−1) generally increased with increasing P concentrations in the growth medium, but growth rate and the ratio of total cell-associated to intracellular P remained generally stable. Intracellular P quotas and growth rates in cells grown under the different P treatments illustrate the ability of this organism to successfully respond to changes in ambient P loads, and thus have implications for ecosystem scale productivity models employing P concentrations to predict algal bloom events. PMID:22279445

Cell culture purity issues and DFAT cells

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

Wei, Shengjuan; Department of Animal Sciences, Washington State University, Pullman, WA 99164; Bergen, Werner G.

2013-04-12

Highlights: •DFAT cells are progeny cells derived from dedifferentiated mature adipocytes. •Common problems in this research is potential cell contamination of initial cultures. •The initial cell culture purity is crucial in DFAT cell research field. -- Abstract: Dedifferentiation of mature adipocytes, in vitro, has been pursued/documented for over forty years. The subsequent progeny cells are named dedifferentiated adipocyte-derived progeny cells (DFAT cells). DFAT cells are proliferative and likely to possess mutilineage potential. As a consequence, DFAT cells and their progeny/daughter cells may be useful as a potential tool for various aspects of tissue engineering and as potential vectors for themore » alleviation of several disease states. Publications in this area have been increasing annually, but the purity of the initial culture of mature adipocytes has seldom been documented. Consequently, it is not always clear whether DFAT cells are derived from dedifferentiated mature (lipid filled) adipocytes or from contaminating cells that reside in an impure culture.« less

Sapphire Energy - Integrated Algal Biorefinery

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

White, Rebecca L.; Tyler, Mike

2015-07-22

Sapphire Energy, Inc. (SEI) is a leader in large-scale photosynthetic algal biomass production, with a strongly cohesive research, development, and operations program. SEI takes a multidiscipline approach to integrate lab-based strain selection, cultivation and harvest and production scale, and extraction for the production of Green Crude oil, a drop in replacement for traditional crude oil.. SEI’s technical accomplishments since 2007 have produced a multifunctional platform that can address needs for fuel, feed, and other higher value products. Figure 1 outlines SEI’s commercialization process, including Green Crude production and refinement to drop in fuel replacements. The large scale algal biomass productionmore » facility, the SEI Integrated Algal Biorefinery (IABR), was built in Luna County near Columbus, New Mexico (see fig 2). The extraction unit was located at the existing SEI facility in Las Cruces, New Mexico, approximately 95 miles from the IABR. The IABR facility was constructed on time and on budget, and the extraction unit expansion to accommodate the biomass output from the IABR was completed in October 2012. The IABR facility uses open pond cultivation with a proprietary harvesting method to produce algal biomass; this biomass is then shipped to the extraction facility for conversion to Green Crude. The operation of the IABR and the extraction facilities has demonstrated the critical integration of traditional agricultural techniques with algae cultivation knowledge for algal biomass production, and the successful conversion of the biomass to Green Crude. All primary unit operations are de-risked, and at a scale suitable for process demonstration. The results are stable, reliable, and long-term cultivation of strains for year round algal biomass production. From June 2012 to November 2014, the IABR and extraction facilities produced 524 metric tons (MT) of biomass (on a dry weight basis), and 2,587 gallons of Green Crude. Additionally, the

The influence of algal densities on the toxicity of chromium for Ceriodaphnia dubia Richard (Cladocera, Crustacea).

PubMed

Rodgher, S; Espíndola, E L G

2008-05-01

Food availability may affect metal toxicity for aquatic organisms. In the present study, the influence of high, medium and low densities of the algae Pseudokirchneriella subcapitata (10(6), 10(5) and 10(4) cells.mL(-1), respectively) on the chronic toxicity of chromium to the cladoceran Ceriodaphnia dubia was investigated. C. dubia was exposed to a range of chromium concentration from 2.71 to 34.04 microg.L(-1) and fed with algae at various densities. In another experiment, the green alga was exposed to chromium concentrations (94 to 774 microg.L(-1)) and supplied as food in different densities to zooplankton. The survival and reproduction of the cladoceran were measured in these toxicity tests. The IC50 for Cr to P. subcapitata and metal accumulated by algal cells were determined. The results of a bifactorial analysis (metal versus algal densities) showed that metal toxicity to zooplankton was dependent on algal densities. Significant toxic effects on the reproduction and survival of C. dubia were observed at 8.73, 18.22 and 34.04 microg.L(-1) Cr when the test organisms were fed with 10(6) cells.mL(-1) of P. subcapitata. Although the chlorophyta retain low chromium content, a decrease in the reproduction and survival of C. dubia occurred when they were fed with high algal density contaminated with 774 microg.L(-1) Cr. It was concluded that high algal density have an appreciable influence on chromium toxicity to daphnids.

Simultaneous Wastewater Treatment, Algal Biomass Production and Electricity Generation in Clayware Microbial Carbon Capture Cells.

PubMed

Jadhav, Dipak A; Jain, Sumat C; Ghangrekar, Makarand M

2017-11-01

Performance of microbial carbon capture cells (MCCs), having a low-cost clayware separator, was evaluated in terms of wastewater treatment and electricity generation using algae Chlorella pyrenoidosa in MCC-1 and Anabaena ambigua in MCC-2 and without algae in a cathodic chamber of MCC-3. Higher power production was achieved in MCC-1 (6.4 W/m 3 ) compared to MCC-2 (4.29 W/m 3 ) and MCC-3 (3.29 W/m 3 ). Higher coulombic efficiency (15.23 ± 1.30%) and biomass production (66.4 ± 4.7 mg/(L*day)) in MCC-1 indicated the superiority of Chlorella over Anabaena algae for carbon capture and oxygen production to facilitate the cathodic reduction. Algal biofilm formation on the cathode surface of MCC-1 increased dissolved oxygen in the catholyte and decreased the cathodic charge transfer resistance with increase in reduction current. Electrochemical analyses revealed slow cathodic reactions and increase in internal resistance in MCC-2 (55 Ω) than MCC-1 (30 Ω), due to lower oxygen produced by Anabaena algae. Thus, biomass production in conjunction with wastewater treatment, CO 2 sequestration and electricity generation can be achieved using Chlorella algal biocathode in MCC.

Methods for removing contaminants from algal oil

DOEpatents

Lupton, Francis Stephen

2016-09-27

Methods for removing contaminants from algal oil are provided. In an embodiment, a method comprises the steps of combining a sulfuric acid-aqueous solution that has a pH of about 1 or less with a contaminant-containing algal oil at treatment conditions effective to form an effluent. The effluent comprises a treated algal oil phase and contaminants in an acidic aqueous phase. The contaminants comprise metals, phosphorus, or combinations thereof. The acidic aqueous phase is removed from the effluent to form a contaminant-depleted algal oil.

Evaluation of Algal Biofilms on Indium Tin Oxide (ITO) for Use in Biophotovoltaic Platforms Based on Photosynthetic Performance

PubMed Central

Ng, Fong-Lee; Phang, Siew-Moi; Periasamy, Vengadesh; Yunus, Kamran; Fisher, Adrian C.

2014-01-01

In photosynthesis, a very small amount of the solar energy absorbed is transformed into chemical energy, while the rest is wasted as heat and fluorescence. This excess energy can be harvested through biophotovoltaic platforms to generate electrical energy. In this study, algal biofilms formed on ITO anodes were investigated for use in the algal biophotovoltaic platforms. Sixteen algal strains, comprising local isolates and two diatoms obtained from the Culture Collection of Marine Phytoplankton (CCMP), USA, were screened and eight were selected based on the growth rate, biochemical composition and photosynthesis performance using suspension cultures. Differences in biofilm formation between the eight algal strains as well as their rapid light curve (RLC) generated using a pulse amplitude modulation (PAM) fluorometer, were examined. The RLC provides detailed information on the saturation characteristics of electron transport and overall photosynthetic performance of the algae. Four algal strains, belonging to the Cyanophyta (Cyanobacteria) Synechococcus elongatus (UMACC 105), Spirulina platensis. (UMACC 159) and the Chlorophyta Chlorella vulgaris (UMACC 051), and Chlorella sp. (UMACC 313) were finally selected for investigation using biophotovoltaic platforms. Based on power output per Chl-a content, the algae can be ranked as follows: Synechococcus elongatus (UMACC 105) (6.38×10−5 Wm−2/µgChl-a)>Chlorella vulgaris UMACC 051 (2.24×10−5 Wm−2/µgChl-a)>Chlorella sp.(UMACC 313) (1.43×10−5 Wm−2/µgChl-a)>Spirulina platensis (UMACC 159) (4.90×10−6 Wm−2/µgChl-a). Our study showed that local algal strains have potential for use in biophotovoltaic platforms due to their high photosynthetic performance, ability to produce biofilm and generation of electrical power. PMID:24874081

Algal Supply System Design - Harmonized Version

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

Abodeely, Jared; Stevens, Daniel; Ray, Allison

2013-03-01

The objective of this design report is to provide an assessment of current technologies used for production, dewatering, and converting microalgae cultivated in open-pond systems to biofuel. The original draft design was created in 2011 and has subsequently been brought into agreement with the DOE harmonized model. The design report extends beyond this harmonized model to discuss some of the challenges with assessing algal production systems, including the ability to (1) quickly assess alternative algal production system designs, (2) assess spatial and temporal variability, and (3) perform large-scale assessments considering multiple scenarios for thousands of potential sites. The Algae Logisticsmore » Model (ALM) was developed to address each of these limitations of current modeling efforts to enable assessment of the economic feasibility of algal production systems across the United States. The (ALM) enables (1) dynamic assessments using spatiotemporal conditions, (2) exploration of algal production system design configurations, (3) investigation of algal production system operating assumptions, and (4) trade-off assessments with technology decisions and operating assumptions. The report discusses results from the ALM, which is used to assess the baseline design determined by harmonization efforts between U.S. DOE national laboratories. Productivity and resource assessment data is provided by coupling the ALM with the Biomass Assessment Tool developed at PNNL. This high-fidelity data is dynamically passed to the ALM and used to help better understand the impacts of spatial and temporal constraints on algal production systems by providing a cost for producing extracted algal lipids annually for each potential site.« less

Direct utilization of waste water algal biomass for ethanol production by cellulolytic Clostridium phytofermentans DSM1183.

PubMed

Fathima, Anwar Aliya; Sanitha, Mary; Kumar, Thangarathinam; Iyappan, Sellamuthu; Ramya, Mohandass

2016-02-01

Direct bioconversion of waste water algal biomass into ethanol using Clostridium phytofermentans DSM1183 was demonstrated in this study. Fermentation of 2% (w/v) autoclaved algal biomass produced ethanol concentration of 0.52 g L(-1) (solvent yield of 0.19 g/g) where as fermentation of acid pretreated algal biomass (2%, w/v) produced ethanol concentration of 4.6 g L(-1) in GS2 media (solvent yield of 0.26 g/g). The control experiment with 2% (w/v) glucose in GS2 media produced ethanol concentration of 2.8 g L(-1) (solvent yield of 0.25 g/g). The microalgal strains from waste water algal biomass were identified as Chlamydomonas dorsoventralis, Graesiella emersonii, Coelastrum proboscideum, Scenedesmus obliquus, Micractinium sp., Desmodesmus sp., and Chlorella sp., based on ITS-2 molecular marker. The presence of glucose, galactose, xylose and rhamnose were detected by high performance liquid chromatography in the algal biomass. Scanning Electron Microscopy observations of fermentation samples showed characteristic morphological changes in algal cells and bioaccessibility of C. phytofermentans. Copyright © 2015 Elsevier Ltd. All rights reserved.

Investigation of animal and algal bioassays for reliable saxitoxin ecotoxicity and cytotoxicity risk evaluation.

PubMed

Perreault, François; Matias, Marcelo Seleme; Melegari, Silvia Pedroso; Pinto, Catia Regina Silva de Carvalho; Creppy, Edmond Ekué; Popovic, Radovan; Matias, William Gerson

2011-05-01

Contamination of water bodies by saxitoxin can result in various toxic effects in aquatic organisms. Saxitoxin contamination has also been shown to be a threat to human health in several reported cases, even resulting in death. In this study, we evaluated the sensitivity of animal (Neuro-2A) and algal (Chlamydomonas reinhardtii) bioassays to saxitoxin effect. Neuro-2A cells were found to be sensitive to saxitoxin, as shown by a 24 h EC50 value of 1.5 nM, which was obtained using a cell viability assay. Conversely, no saxitoxin effect was found in any of the algal biomarkers evaluated, for the concentration range tested (2-128 nM). These results indicate that saxitoxin may induce toxic effects in animal and human populations at concentrations where phytoplankton communities are not affected. Therefore, when evaluating STX risk of toxicity, algal bioassays do not appear to be reliable indicators and should always be conducted in combination with animal bioassays. Copyright © 2011 Elsevier Inc. All rights reserved.

Interactions among irradiance, nutrients, and herbivores constrain a stream algal community.

PubMed

Rosemond, A D

1993-07-01

Using stream-side, flow-through channels, I tested for the effects of nutrients (NU) (nitrogen plus phosphorus), irradiance (L), and snail grazing (G) on a benthic algal community in a small, forested stream. Grazed communities were-dominated by a chlorophyte (basal cells ofStigeoclonium) and a cyanophyte (Chamaesiphon investiens), whereas ungrazed communities were comprised almost entirely of diatoms, regardless of nutrient and light levels. Snails maintained low algal biomass in all grazed treatments, presumably by consuming increased algal production in treatments to which L and NU were increased. When nutrients were increased, cellular nutrient content increased under ambient conditions (shaded, grazed) and biomass and productivity increased when snails were removed and light was increased. Together, nutrients and light had positive effects and grazing had negative effects on biomass (chlorophylla, AFDM, algal biovolume) and chlorophyll-and areal-specific productivity in ANOVAs. However, in most cases, only means from treatments in which all three factors were manipulated (ungrazed, +NU&L treatments) were significantly different from controls; effects of single factors were generally undetectable. These results indicate that all three factors simultaneously limited algal biomass and productivity in this stream during the summer months. Additionally, the effects of these factors in combination were in some cases different from the effects of single factors. For example, light had slight negative effects on some biomass parameters when added at ambient snail densities and nutrient concentrations, but had strong positive effects in conjunction with nutrient addition and snail removal. This study demonstrates that algal biomass and productivity can be under multiple constraints by irradiance, nutrients, and herbivores and indicates the need to employ multifactor experiments to test for such interactive effects.

Control of algal production in a high rate algal pond: investigation through batch and continuous experiments.

PubMed

Derabe Maobe, H; Onodera, M; Takahashi, M; Satoh, H; Fukazawa, T

2014-01-01

For decades, arid and semi-arid regions in Africa have faced issues related to water availability for drinking, irrigation and livestock purposes. To tackle these issues, a laboratory scale greywater treatment system based on high rate algal pond (HRAP) technology was investigated in order to guide the operation of the pilot plant implemented in the 2iE campus in Ouagadougou (Burkina Faso). Because of the high suspended solids concentration generally found in effluents of this system, the aim of this study is to improve the performance of HRAPs in term of algal productivity and removal. To determine the selection mechanism of self-flocculated algae, three sets of sequencing batch reactors (SBRs) and three sets of continuous flow reactors (CFRs) were operated. Despite operation with the same solids retention time and the similarity of the algal growth rate found in these reactors, the algal productivity was higher in the SBRs owing to the short hydraulic retention time of 10 days in these reactors. By using a volume of CFR with twice the volume of our experimental CFRs, the algal concentration can be controlled during operation under similar physical conditions in both reactors.

Water-quality parameters and benthic algal communities at selected streams in Minnesota, August 2000 - Study design, methods and data

USGS Publications Warehouse

Lee, K.E.

2002-01-01

This report describes the study design, sampling methods, and summarizes the physical, chemical, and benthic algal data for a component of the multiagency study that was designed to document diurnal water-quality measurements (specific conductance, pH, water temperature, and dissolved oxygen), benthic algal community composition and chlorophyll-a content, and primary productivity at 12 stream sites on 6 streams in Minnesota during August 2000. Specific conductance, pH, water temperature, dissolved oxygen concentrations and percent dissolved oxygen saturation measurements were made with submersible data recorders at 30 minute intervals for a period of 3-6 days during August 2000. Benthic algae collected from wood and rock substrate were identified and enumerated. Biovolume (volume of algal cells per unit area), density (number of cells per unit area), and chlorophyll-a content from benthic algae were determined. These data can be used as part of the multiagency study to develop an understanding of the relations among nutrient concentrations, algal abundance, algal community composition, and primary production and respiration processes in rivers of differing ecoregions in Minnesota.

Optimization of culture conditions and medium composition for the marine algicidal bacterium Alteromonas sp. DH46 by uniform design

NASA Astrophysics Data System (ADS)

Lin, Jing; Zheng, Wei; Tian, Yun; Wang, Guizhong; Zheng, Tianling

2013-09-01

Harmful algal blooms (HABs) have led to extensive ecological and environmental issues and huge economic losses. Various HAB control techniques have been developed, and biological methods have been paid more attention. Algicidal bacteria is a general designation for bacteria which inhibit algal growth in a direct or indirect manner, and kill or damage the algal cells. A metabolite which is strongly toxic to the dinoflagellate Alexandrium tamarense was produced by strain DH46 of the alga-lysing bacterium Alteromonas sp. The culture conditions were optimized using a single-factor test method. Factors including carbon source, nitrogen source, temperature, initial pH value, rotational speed and salinity were studied. The results showed that the cultivation of the bacteria at 28°C and 180 r min-1 with initial pH 7 and 30 salt contcentration favored both the cell growth and the lysing effect of strain DH46. The optimal medium composition for strain DH46 was determined by means of uniform design experimentation, and the most important components influencing the cell density were tryptone, yeast extract, soluble starch, NaNO3 and MgSO4. When the following culture medium was used (tryptone 14.0g, yeast extract 1.63g, soluble starch 5.0 g, NaNO3 1.6 g, MgSO4 2.3 g in 1L), the largest bacterial dry weight (7.36 g L-1) was obtained, which was an enhancement of 107% compared to the initial medium; and the algal lysis rate was as high as 98.4% which increased nearly 10% after optimization.

Mammalian Cell Tissue Culture.

PubMed

Phelan, Katy; May, Kristin M

2017-07-11

Cultured mammalian cells are used extensively in the field of human genetics. It requires a number of special skills in order to be able to preserve the structure, function, behavior, and biology of the cells in culture. This unit describes the basic skills required to maintain and preserve cell cultures: maintaining aseptic technique, preparing media with the appropriate characteristics, passaging, freezing and storage, recovering frozen stocks, and counting viable cells. © 2017 by John Wiley & Sons, Inc. Copyright © 2017 John Wiley & Sons, Inc.

Dynamics of Bacterial and Fungal Communities during the Outbreak and Decline of an Algal Bloom in a Drinking Water Reservoir.

PubMed

Zhang, Haihan; Jia, Jingyu; Chen, Shengnan; Huang, Tinglin; Wang, Yue; Zhao, Zhenfang; Feng, Ji; Hao, Huiyan; Li, Sulin; Ma, Xinxin

2018-02-18

The microbial communities associated with algal blooms play a pivotal role in organic carbon, nitrogen and phosphorus cycling in freshwater ecosystems. However, there have been few studies focused on unveiling the dynamics of bacterial and fungal communities during the outbreak and decline of algal blooms in drinking water reservoirs. To address this issue, the compositions of bacterial and fungal communities were assessed in the Zhoucun drinking water reservoir using 16S rRNA and internal transcribed spacer (ITS) gene Illumina MiSeq sequencing techniques. The results showed the algal bloom was dominated by Synechococcus, Microcystis, and Prochlorothrix. The bloom was characterized by a steady decrease of total phosphorus (TP) from the outbreak to the decline period (p < 0.05) while Fe concentration increased sharply during the decline period (p < 0.05). The highest algal biomass and cell concentrations observed during the bloom were 51.7 mg/L and 1.9×108 cell/L, respectively. The cell concentration was positively correlated with CODMn (r = 0.89, p = 0.02). Illumina Miseq sequencing showed that algal bloom altered the water bacterial and fungal community structure. During the bloom, the dominant bacterial genus were Acinetobacter sp., Limnobacter sp., Synechococcus sp., and Roseomonas sp. The relative size of the fungal community also changed with algal bloom and its composition mainly contained Ascomycota, Basidiomycota and Chytridiomycota. Heat map profiling indicated that algal bloom had a more consistent effect upon fungal communities at genus level. Redundancy analysis (RDA) also demonstrated that the structure of water bacterial communities was significantly correlated to conductivity and ammonia nitrogen. Meanwhile, water temperature, Fe and ammonia nitrogen drive the dynamics of water fungal communities. The results from this work suggested that water bacterial and fungal communities changed significantly during the outbreak and decline of algal bloom in

Dynamics of Bacterial and Fungal Communities during the Outbreak and Decline of an Algal Bloom in a Drinking Water Reservoir

PubMed Central

Zhang, Haihan; Jia, Jingyu; Chen, Shengnan; Huang, Tinglin; Wang, Yue; Zhao, Zhenfang; Feng, Ji; Hao, Huiyan; Li, Sulin; Ma, Xinxin

2018-01-01

The microbial communities associated with algal blooms play a pivotal role in organic carbon, nitrogen and phosphorus cycling in freshwater ecosystems. However, there have been few studies focused on unveiling the dynamics of bacterial and fungal communities during the outbreak and decline of algal blooms in drinking water reservoirs. To address this issue, the compositions of bacterial and fungal communities were assessed in the Zhoucun drinking water reservoir using 16S rRNA and internal transcribed spacer (ITS) gene Illumina MiSeq sequencing techniques. The results showed the algal bloom was dominated by Synechococcus, Microcystis, and Prochlorothrix. The bloom was characterized by a steady decrease of total phosphorus (TP) from the outbreak to the decline period (p < 0.05) while Fe concentration increased sharply during the decline period (p < 0.05). The highest algal biomass and cell concentrations observed during the bloom were 51.7 mg/L and 1.9×108 cell/L, respectively. The cell concentration was positively correlated with CODMn (r = 0.89, p = 0.02). Illumina Miseq sequencing showed that algal bloom altered the water bacterial and fungal community structure. During the bloom, the dominant bacterial genus were Acinetobacter sp., Limnobacter sp., Synechococcus sp., and Roseomonas sp. The relative size of the fungal community also changed with algal bloom and its composition mainly contained Ascomycota, Basidiomycota and Chytridiomycota. Heat map profiling indicated that algal bloom had a more consistent effect upon fungal communities at genus level. Redundancy analysis (RDA) also demonstrated that the structure of water bacterial communities was significantly correlated to conductivity and ammonia nitrogen. Meanwhile, water temperature, Fe and ammonia nitrogen drive the dynamics of water fungal communities. The results from this work suggested that water bacterial and fungal communities changed significantly during the outbreak and decline of algal bloom in

Lysing bloom-causing alga Phaeocystis globosa with microbial algicide: An efficient process that decreases the toxicity of algal exudates

PubMed Central

Cai, Guanjing; Yang, Xujun; Lai, Qiliang; Yu, Xiaoqi; Zhang, Huajun; Li, Yi; Chen, Zhangran; Lei, Xueqian; Zheng, Wei; Xu, Hong; Zheng, Tianling

2016-01-01

Algicidal microbes could effectively remove the harmful algae from the waters. In this study, we were concerned with the ecological influence of an algicide extracted from Streptomyces alboflavus RPS, which could completely lyse the Phaeocystis globosa cells within two days. In microcosms, 4 μg/mL of the microbial algicide could efficiently remove P. globosa cells without suppressing other aquatic organisms. Bioluminescent assays confirmed that the toxicity of microbial algicide at this concentration was negligible. Interestingly, the toxicity of P. globosa exudates was also significantly reduced after being treated with the algicide. Further experiments revealed that the microbial algicide could instantly increase the permeability of the plasma membrane and disturb the photosynthetic system, followed by the deformation of organelles, vacuolization and increasing oxidative stress. The pre-incubation of N-acetyl cysteine (NAC) verified that the rapid damages to the plasma membrane and photosynthetic system caused the algal death in the early phase, and the increasing oxidative stress killed the rest. The late accumulation and possible release of CAT also explained the decreasing toxicity of the algal culture. These results indicated that this microbial algicide has great potential in controlling the growth of P. globosa on site. PMID:26847810

Fish Stem Cell Cultures

PubMed Central

Hong, Ni; Li, Zhendong; Hong, Yunhan

2011-01-01

Stem cells have the potential for self-renewal and differentiation. First stem cell cultures were derived 30 years ago from early developing mouse embryos. These are pluripotent embryonic stem (ES) cells. Efforts towards ES cell derivation have been attempted in other mammalian and non-mammalian species. Work with stem cell culture in fish started 20 years ago. Laboratory fish species, in particular zebrafish and medaka, have been the focus of research towards stem cell cultures. Medaka is the second organism that generated ES cells and the first that gave rise to a spermatogonial stem cell line capable of test-tube sperm production. Most recently, the first haploid stem cells capable of producing whole animals have also been generated from medaka. ES-like cells have been reported also in zebrafish and several marine species. Attempts for germline transmission of ES cell cultures and gene targeting have been reported in zebrafish. Recent years have witnessed the progress in markers and procedures for ES cell characterization. These include the identification of fish homologs/paralogs of mammalian pluripotency genes and parameters for optimal chimera formation. In addition, fish germ cell cultures and transplantation have attracted considerable interest for germline transmission and surrogate production. Haploid ES cell nuclear transfer has proven in medaka the feasibility of semi-cloning as a novel assisted reproductive technology. In this special issue on “Fish Stem Cells and Nuclear Transfer”, we will focus our review on medaka to illustrate the current status and perspective of fish stem cells in research and application. We will also mention semi-cloning as a new development to conventional nuclear transfer. PMID:21547056

Fish stem cell cultures.

PubMed

Hong, Ni; Li, Zhendong; Hong, Yunhan

2011-04-13

Stem cells have the potential for self-renewal and differentiation. First stem cell cultures were derived 30 years ago from early developing mouse embryos. These are pluripotent embryonic stem (ES) cells. Efforts towards ES cell derivation have been attempted in other mammalian and non-mammalian species. Work with stem cell culture in fish started 20 years ago. Laboratory fish species, in particular zebrafish and medaka, have been the focus of research towards stem cell cultures. Medaka is the second organism that generated ES cells and the first that gave rise to a spermatogonial stem cell line capable of test-tube sperm production. Most recently, the first haploid stem cells capable of producing whole animals have also been generated from medaka. ES-like cells have been reported also in zebrafish and several marine species. Attempts for germline transmission of ES cell cultures and gene targeting have been reported in zebrafish. Recent years have witnessed the progress in markers and procedures for ES cell characterization. These include the identification of fish homologs/paralogs of mammalian pluripotency genes and parameters for optimal chimera formation. In addition, fish germ cell cultures and transplantation have attracted considerable interest for germline transmission and surrogate production. Haploid ES cell nuclear transfer has proven in medaka the feasibility of semi-cloning as a novel assisted reproductive technology. In this special issue on "Fish Stem Cells and Nuclear Transfer", we will focus our review on medaka to illustrate the current status and perspective of fish stem cells in research and application. We will also mention semi-cloning as a new development to conventional nuclear transfer.

Lipid production of Chlorella vulgaris cultured in artificial wastewater medium.

PubMed

Feng, Yujie; Li, Chao; Zhang, Dawei

2011-01-01

Chlorella vulgaris was used to study algal lipid production with wastewater treatment. Artificial wastewater was used to cultivate C. vulgaris in a column aeration photobioreactor (CAP) under batch and semi-continuous cultivation with various daily culture replacements (0.5l-1.5l per 2l reactor). The cell density was decreased from 0.89 g/l with the daily replacement of 0.5l to 0.28 g/l with 1.5l replacement. However, C. vulgaris culture achieved the highest lipid content (42%, average value of the phase) and the lipid productivity (147 mg/ld(-1)) with daily replacement of 1.0 l. And then the nutrient removal efficiency were 86% (COD), 97% (NH(4)(+)) and 96% (TP), respectively. Analyses of energy efficiency showed that the net energy ratio (NER) for lipid production with daily replacement of 1.0 l (1.25) was higher than the other volume replacement protocols. And cost analyses showed that the algal biomass can be competitive with petroleum at US$ 63.97 per barrel with the potential credit for wastewater treatment. According to the above results, it is concluded that the present research will lead to an economical technology of algal lipid production. Copyright © 2010 Elsevier Ltd. All rights reserved.

Problems related to water quality and algal control in Lopez Reservoir, San Luis Obispo County, California

USGS Publications Warehouse

Fuller, Richard H.; Averett, Robert C.; Hines, Walter G.

1975-01-01

A study to determine the present enrichment status of Liopez Reservoir in San Luis Obispo county, California, and to evaluate copper sulfate algal treatment found that stratification in the reservoir regulates nutrient release and that algal control has been ineffective. Nuisance algal blooms, particularly from March to June, have been a problem in the warm multipurpose reservoir since it was initially filled following intense storms in 1968-69. The cyanophyte Anabaena unispora has been dominant; cospecies are the diatoms Stephanodiscus astraea and Cyclotella operculata, and the chlorophytes Pediastrum deplex and Sphaerocystis schroeteri. During an A. unispora bloom in May 1972 the total lake surface cell count was nearly 100,000 cells/ml. Thermal stratification from late spring through autumn results in oxygen deficiency in the hypolimnion and metalimnion caused by bacterial oxidation of organic detritus. The anaerobic conditions favor chemical reduction of organic matter, which constitute 10-14% of the sediment. As algae die, sink to the bottom, and decompose, nutrients are released to the hypolimnion , and with the autumn overturn are spread to the epilimnion. Algal blooms not only hamper recreation, but through depletion of dissolved oxygen in the epilimnion may have caused periodic fishkills. Copper sulfate mixed with sodium citrate and applied at 1.10-1.73 lbs/acre has not significantly reduced algal growth; a method for determining correct dosage is presented. (Lynch-Wisconsin)

Reeling in the damages: Harmful algal blooms' impact on Lake Erie's recreational fishing industry.

PubMed

Wolf, David; Georgic, Will; Klaiber, H Allen

2017-09-01

Lake Erie is one of the most valuable natural resources in the United States, providing billions of dollars in benefits each year to recreationalists, homeowners and local governments. The ecosystem services provided by Lake Erie, however, are under threat due to harmful algal blooms. This paper provides recreational damage estimates using spatially and temporally varying algae measures and monthly fishing permit sales collected between 2011 and 2014. Results indicate that fishing license sales drop between 10% and 13% when algal conditions surpass the World Health's Organization's moderate health risk advisory threshold of 20,000 cyanobacteria cells/mL. For Lake Erie adjacent counties experiencing a large, summer-long algal bloom, this would result in approximately 3600 fewer fishing licenses issued and approximately $2.25 million to $5.58 million in lost fishing expenditures. Our results show a discrete jump in reduced angling activity upon crossing this threshold, with limited additional impacts associated with more severe algal blooms. This suggests that policies aimed at eliminating, rather than mitigating, algal levels are most beneficial to the Ohio angling industry. Copyright © 2017 Elsevier Ltd. All rights reserved.

Pumps for microfluidic cell culture.

PubMed

Byun, Chang Kyu; Abi-Samra, Kameel; Cho, Yoon-Kyoung; Takayama, Shuichi

2014-02-01

In comparison to traditional in vitro cell culture in Petri dishes or well plates, cell culture in microfluidic-based devices enables better control over chemical and physical environments, higher levels of experimental automation, and a reduction in experimental materials. Over the past decade, the advantages associated with cell culturing in microfluidic-based platforms have garnered significant interest and have led to a plethora of studies for high throughput cell assays, organs-on-a-chip applications, temporal signaling studies, and cell sorting. A clear concern for performing cell culture in microfluidic-based devices is deciding on a technique to deliver and pump media to cells that are encased in a microfluidic device. In this review, we summarize recent advances in pumping techniques for microfluidic cell culture and discuss their advantages and possible drawbacks. The ultimate goal of our review is to distill the large body of information available related to pumps for microfluidic cell culture in an effort to assist current and potential users of microfluidic-based devices for advanced in vitro cellular studies. © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Behavior and function of paternally inherited centrioles in brown algal zygotes.

PubMed

Nagasato, Chikako

2005-12-01

In brown algal cells, the centrosome, consisting of a pair of centrioles and the pericentriolar material, is primarily involved in the organization of microtubules (MTs) throughout the cell cycle. In motile cells, the centrioles participate in the formation of flagellar axoneme as flagellar basal bodies, and in somatic cells they play a crucial role in many cellular activities as a part of the centrosome. With respect to the role of the centrosome as a microtubule organizing center (MTOC), brown algal cells resemble animal cells. In most animal fertilization processes, the sperm cell introduces centrioles, the core of the centrosome, into the egg cytoplasm. In this study, the behavior of centrioles from gametogenesis and fertilization to the first cell division of the zygote was examined in the three sexual reproduction patterns occurring in brown algae, i.e., oogamy, anisogamy and isogamy, by electron- and immunofluorescence-microscopy. The pair of centrioles contained in somatic cells was shown to be derived from the male gamete, irrespective of the sexual reproductive pattern. The paternally derived centrioles were duplicated before mitosis and were involved in spindle pole formation. Moreover, MTs from the centrosome play a crucial role in the process of cytokinesis, as the position of centrosomes accompanying daughter nuclei seems to determine the cytokinetic plane. A new approach to clarifying the mode of cytokinesis in brown algae is presented in this study.

Recent developments on algal biochar production and characterization.

PubMed

Yu, Kai Ling; Lau, Beng Fye; Show, Pau Loke; Ong, Hwai Chyuan; Ling, Tau Chuan; Chen, Wei-Hsin; Ng, Eng Poh; Chang, Jo-Shu

2017-12-01

Algal biomass is known as a promising sustainable feedstock for the production of biofuels and other valuable products. However, since last decade, massive amount of interests have turned to converting algal biomass into biochar. Due to their high nutrient content and ion-exchange capacity, algal biochars can be used as soil amendment for agriculture purposes or adsorbents in wastewater treatment for the removal of organic or inorganic pollutants. This review describes the conventional (e.g., slow and microwave-assisted pyrolysis) and newly developed (e.g., hydrothermal carbonization and torrefaction) methods used for the synthesis of algae-based biochars. The characterization of algal biochar and a comparison between algal biochar with biochar produced from other feedstocks are also presented. This review aims to provide updated information on the development of algal biochar in terms of the production methods and the characterization of its physical and chemical properties to justify and to expand their potential applications. Copyright © 2017 Elsevier Ltd. All rights reserved.

Production of ω-3 Polyunsaturated Fatty Acids From Cull Potato Using an Algae Culture Process

NASA Astrophysics Data System (ADS)

Chi, Zhanyou; Hu, Bo; Liu, Yan; Frear, Craig; Wen, Zhiyou; Chen, Shulin

Algal cultivation for converting cull potato to docosahexaenoic acid (DHA) was studied. Schizochytrium limacinum SR21 was selected as the better producing strain, compared with Thraustochytrium aureum because of higher cell density and DHA content. Used as both carbon and nitrogen source, an optimal ratio of hydrolyzed potato broth in the culture medium was determined as 50%, with which the highest production of 21.7 g/L dry algae biomass and 5.35 g/L DHA was obtained, with extra glucose supplemented. Repeat culture further improved the cell density but not fed batch culture, suggesting limited growth was most likely caused by metabolites inhibition.

Cell Culture Made Easy.

ERIC Educational Resources Information Center

Dye, Frank J.

1985-01-01

Outlines steps to generate cell samples for observation and experimentation. The procedures (which use ordinary laboratory equipment) will establish a short-term primary culture of normal mammalian cells. Information on culture vessels and cell division and a list of questions to generate student interest and involvement in the topics are…

NREL Algal Biofuels Projects and Partnerships

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

2016-10-01

This fact sheet highlights several algal biofuels research and development projects focused on improving the economics of the algal biofuels production process. These projects should serve as a foundation for the research efforts toward algae as a source of fuels and other chemicals.

Demography of zooplankton (Anuraeopsis fissa, Brachionus rubens and Moina macrocopa) fed Chlorella vulgaris and Scenedesmus acutus cultured on different media.

PubMed

Morales-Ventura, Jesús; Nandini, S; Sarma, S S S; Castellanos-Páez, Maria Elena

2012-09-01

Generally zooplankton growth is often limited by the quality of their algal diet. A cheaper common practice in aquaculture, is to culture algae with fertilizers; however, the demography of zooplankton when fed these algae has not yet been evaluated. We studied the population growth and life table demography of the rotifers Anuraeopsis fissa and Brachionus rubens, and the cladoceran Moina macrocopa. For this, the algae Scenedesmus acutus or Chlorella vulgaris were cultured on defined (Bold's basal) medium or the commercial liquid fertilizer (Bayfolan). Experiments were conducted at one algal concentration 1.0 x 10(6) cells/mL of C. vulgaris or its equivalent dry weight of 0.5 x 10(6) cells/mL of S. acutus. The population dynamics were tested at 23 +/- 1 degrees C in 100 mL transparent jars, each with 50mL of the test medium, with an initial density of 0.5indiv/mL, for a total of 48 test jars (3 zooplankton 2 algal species x 2 culture media x 4 replicates). For the life table experiments with M. macrocopa, we introduced 10 neonates (<24h old) into each test jar containing the specific algal type and concentration. For the rotifer experiments, we set 5mL tubes with one neonate each and 10 replicates for each algal species and culture medium. We found that the average rotifer life span was not influenced by the diet, but for M. macrocopa fed S. acutus cultured in Bold's medium, the average lifespan was significantly lower than with the other diets. The gross and net reproductive rates of A. fissa (ranging from 18-36 offspring per female) were significantly higher for C vulgaris cultured in Bold medium. Regardless of the culture medium, Chlorella resulted in significantly higher gross and net reproductive rates for B. rubens than S. acutus diets. The reproductive rates of M. macrocopa were significantly higher in all the tested diets except when fed with S. acutus in Bold medium. The population increase rate, derived from growth experiments of A. fissa and B. rubens

Exploration of the antioxidant system and photosynthetic system of a marine algicidal Bacillus and its effect on four harmful algal bloom species.

PubMed

Hou, Shaoling; Shu, Wanjiao; Tan, Shuo; Zhao, Ling; Yin, Pinghe

2016-01-01

A novel marine bacterium, strain B1, initially showed 96.4% algicidal activity against Phaeocystis globosa. Under this situation, 3 other harmful algal species (Skeletonema costatum, Heterosigma akashiwo, and Prorocentrum donghaiense) were chosen to study the algicidal effects of strain B1, and the algicidal activities were 91.4%, 90.7%, and 90.6%, respectively. To explore the algicidal mechanism of strain B1 on these 4 harmful algal species, the characteristics of the antioxidant system and photosynthetic system were studied. Sensitivity to strain B1 supernatant, enzyme activity, and gene expression varied with algal species, while the algicidal patterns were similar. Strain B1 supernatant increased malondialdehyde contents; decreased chlorophyll a contents; changed total antioxidant and superoxide dismutase activity; and restrained psbA, psbD, and rbcL genes expression, which eventually resulted in the algal cells death. The algicidal procedure was observed using field emission scanning electron microscopy, which indicated that algal cells were lysed and cellular substances were released. These findings suggested that the antioxidant and photosynthetic system of these 4 algal species was destroyed under strain B1 supernatant stress. This is the first report to explore and compare the mechanism of a marine Bacillus against harmful algal bloom species of covered 4 phyla.

Henrietta Lacks, HeLa cells, and cell culture contamination.

PubMed

Lucey, Brendan P; Nelson-Rees, Walter A; Hutchins, Grover M

2009-09-01

Henrietta Lacks died in 1951 of an aggressive adenocarcinoma of the cervix. A tissue biopsy obtained for diagnostic evaluation yielded additional tissue for Dr George O. Gey's tissue culture laboratory at Johns Hopkins (Baltimore, Maryland). The cancer cells, now called HeLa cells, grew rapidly in cell culture and became the first human cell line. HeLa cells were used by researchers around the world. However, 20 years after Henrietta Lacks' death, mounting evidence suggested that HeLa cells contaminated and overgrew other cell lines. Cultures, supposedly of tissues such as breast cancer or mouse, proved to be HeLa cells. We describe the history behind the development of HeLa cells, including the first published description of Ms Lacks' autopsy, and the cell culture contamination that resulted. The debate over cell culture contamination began in the 1970s and was not harmonious. Ultimately, the problem was not resolved and it continues today. Finally, we discuss the philosophical implications of the immortal HeLa cell line.

Mammalian Cell Tissue Culture Techniques.

PubMed

Phelan, Katy; May, Kristin M

2016-06-01

Cultured tissues and cells are used extensively in physiological and pharmacological studies. In vitro cultures provide a means of examining cells and tissues without the complex interactions that would be present if the whole organism were studied. A number of special skills are required in order to preserve the structure, function, behavior, and biology of cells in culture. This unit describes the basic skills required to maintain and preserve cell cultures: maintaining aseptic technique, preparing media with the appropriate characteristics, passaging, freezing and storage, recovering frozen stocks, and counting viable cells. © 2016 by John Wiley & Sons, Inc. Copyright © 2016 John Wiley & Sons, Inc.

Development of a novel artificial medium based on utilization of algal photosynthetic metabolites by symbiotic heterotrophs.

PubMed

Watanabe, K; Imase, M; Aoyagi, H; Ohmura, N; Saiki, H; Tanaka, H

2008-09-01

(i) Quantitative and qualitative analyses of photosynthetic metabolites of Chlorella sorokiniana and elucidation of the mechanism of their utilization by algal symbionts. (ii) Development of artificial medium that imitates photoautotroph-heterotroph interaction and investigation of its suitability for isolation of novel microbes from the environment. Various components, including free dissolved carbohydrates, nitrogenous compounds and vitamin, were detected and together contributed 11.1% (as carbon content) of the total photosynthetic metabolites in the medium. Utilization of these photosynthetic metabolites in algal culture broth by algal symbionts was studied. Many symbionts showed specific utilization patterns. A novel artificial extracellular released organic carbon medium, which imitated the nutritional conditions surrounding algae, was developed based on the pattern of utilization of the algal metabolites by the symbiotic heterotrophs. About 42.9% of the isolates were closely related to photoautotrophic-dependent and oligotrophic bacteria. With the novel artificial medium, it was possible to selectively isolate some bacterial strains. Synthetic bacterial growth medium is an important and basic tool for bacterial isolation from environmental samples. The current study shows that preferential separation of typical bacterial subset can be achieved by using artificial medium that mimics photosynthetic metabolites.

Possible importance of algal toxins in the Salton Sea, California

USGS Publications Warehouse

Reifel, K.M.; McCoy, M.P.; Rocke, T.E.; Tiffany, M.A.; Hurlbert, S.H.; Faulkner, D.J.

2002-01-01

In response to wildlife mortality including unexplained eared grebe (Podiceps nigricollis) die-off events in 1992 and 1994 and other mortality events including large fish kills, a survey was conducted for the presence of algal toxins in the Salton Sea. Goals of this survey were to determine if and when algal toxins are present in the Salton Sea and to describe the phytoplankton composition during those times. A total of 29 samples was collected for toxicity analysis from both nearshore and midlake sites visited biweekly from January to December 1999. Dinoflagellates and diatoms dominated most samples, but some were dominated by a prymnesiophyte (Pleurochrysis pseudoroscoffensis) or a raphidophyte (Chattonella marina). Several types of blooms were observed and sampled. The dinoflagellate Gyrodinium uncatenum formed an extensive, dense (up to 310 000 cells ml−1) and long-lasting bloom during the winter in 1999. A coccolithophorid, Pleurochrysis pseudoroscoffensis, occurred at high densities in surface films and nearshore areas during the spring and summer of 1999. These surface films also contained high densities of one or two other species (an unidentified scrippsielloid, Heterocapsa niei, Chattonella marina). Localized blooms were also observed in the Salton Sea. An unknown small dinoflagellate reached high densities (110 000 cells ml−1) inside Varner Harbor, and an unidentified species of Gymnodinium formed a dense (270 000 cells ml−1) band along part of the southern shoreline during the summer. Three species known to produce toxins in other systems were found. Protoceratium reticulatum (=Gonyaulax grindleyi) and Chattonella marina were found in several samples taken during summer months, and Prorocentrum minimum was found in low densities in several samples. Extracts of most samples, including those containing known toxic species, showed a low level (<10% mortality across all concentrations) of activity in the brine shrimp lethality assay

Detection of surface algal blooms using the newly developed algorithm surface algal bloom index (SABI)

NASA Astrophysics Data System (ADS)

Alawadi, Fahad

2010-10-01

Quantifying ocean colour properties has evolved over the past two decades from being able to merely detect their biological activity to the ability to estimate chlorophyll concentration using optical satellite sensors like MODIS and MERIS. The production of chlorophyll spatial distribution maps is a good indicator of plankton biomass (primary production) and is useful for the tracing of oceanographic currents, jets and blooms, including harmful algal blooms (HABs). Depending on the type of HABs involved and the environmental conditions, if their concentration rises above a critical threshold, it can impact the flora and fauna of the aquatic habitat through the introduction of the so called "red tide" phenomenon. The estimation of chlorophyll concentration is derived from quantifying the spectral relationship between the blue and the green bands reflected from the water column. This spectral relationship is employed in the standard ocean colour chlorophyll-a (Chlor-a) product, but is incapable of detecting certain macro-algal species that float near to or at the water surface in the form of dense filaments or mats. The ability to accurately identify algal formations that sometimes appear as oil spill look-alikes in satellite imagery, contributes towards the reduction of false-positive incidents arising from oil spill monitoring operations. Such algal formations that occur in relatively high concentrations may experience, as in land vegetation, what is known as the "red-edge" effect. This phenomena occurs at the highest reflectance slope between the maximum absorption in the red due to the surrounding ocean water and the maximum reflectance in the infra-red due to the photosynthetic pigments present in the surface algae. A new algorithm termed the surface algal bloom index (SABI), has been proposed to delineate the spatial distributions of floating micro-algal species like for example cyanobacteria or exposed inter-tidal vegetation like seagrass. This algorithm was

Renewable sustainable biocatalyzed electricity production in a photosynthetic algal microbial fuel cell (PAMFC).

PubMed

Strik, David P B T B; Terlouw, Hilde; Hamelers, Hubertus V M; Buisman, Cees J N

2008-12-01

Electricity production via solar energy capturing by living higher plants and microalgae in combination with microbial fuel cells are attractive because these systems promise to generate useful energy in a renewable, sustainable, and efficient manner. This study describes the proof of principle of a photosynthetic algal microbial fuel cell (PAMFC) based on naturally selected algae and electrochemically active microorganisms in an open system and without addition of instable or toxic mediators. The developed solar-powered PAMFC produced continuously over 100 days renewable biocatalyzed electricity. The sustainable performance of the PAMFC resulted in a maximum current density of 539 mA/m2 projected anode surface area and a maximum power production of 110 mW/m2 surface area photobioreactor. The energy recovery of the PAMFC can be increased by optimization of the photobioreactor, by reducing the competition from non-electrochemically active microorganisms, by increasing the electrode surface and establishment of a further-enriched biofilm. Since the objective is to produce net renewable energy with algae, future research should also focus on the development of low energy input PAMFCs. This is because current algae production systems have energy inputs similar to the energy present in the outcoming valuable products.

Algal succession and chronosequences on abandoned mine spoils

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

Shubert, L.E.; Starks, T.L.

1978-06-01

Soils were collected from spoil material aged 0 to 45 years. The soils were analyzed for the presence of algal species, chlorophyll ..cap alpha.., major cations, anions and trace elements. There was a gradual increase in the number of algal species and chlorophyll ..cap alpha.. from 1 year old spoils to adjacent unmined natural sites. A total of 41 algal species were identified from all sites. Several species were only found at the unmined sites and they may represent a stable algal community. Results of a statistical analysis on the litho- and chronosequence of the soils will be discussed.

Algal refossilization of atmospheric carbon dioxide. [Contains bibliography

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

Neushul, M.

1991-07-01

The atmospheric concentration of carbon dioxide (CO{sub 2}) is steadily increasing. With our increasing awareness of the economic and environmental impacts of the greenhouse effects'' of CO{sub 2}, methane and other gases, there is interest in finding new methods to reduce the amounts of these gases in the atmosphere. This study evaluates the possibility that large-scale oceanic cultures of macroalgae (macroscopic seaweeds'') could be used to capture atmospheric CO{sub 2}. It is a design for a marine farm system in which a crop'' of calcareous macroalgae grows attached to, and supported by, floating macroalgae that comprise the farm structure.'' Themore » least complicated, yet feasible, macroalgal farm system appears to be one in which laboratory-propagated calcareous algal epiphytes'' and floating algal basiphytes'' are dispersed together in natural ocean upwelling regions. From there, the plants drift with surface currents to the open ocean and then sink to the sea floor, where the buried carbon is refossilized.'' An important caveat regarding the use of calcareous algae is that the process of calcification may release CO{sub 2} to the atmosphere. There is some evidence that CO{sub 2} is not released by calcification in red calcareous algae, but in contrast many geochemists feel that all biologically -- as well as chemically --mediated calcification processes release CO{sub 2}. A substantial amount of research will be necessary to answer basic questions about algal carbon fixation and biomineralization on one hand, while on the other hand to devise strategies for farming the open ocean. 76 refs., 14 figs., 7 tabs.« less

Effects of algal-derived carbon on sediment methane ...

EPA Pesticide Factsheets

Nutrient loading is known to have adverse consequences for aquatic ecosystems, particularly in the form of algal blooms that may result. These blooms pose problems for humans and wildlife, including harmful toxin release, aquatic hypoxia and increased costs for water treatment. Another potential disservice resulting from algal blooms is the enhanced production of methane (CH4), a potent greenhouse gas, in aquatic sediments. Laboratory experiments have shown that algal biomass additions to sediment cores increase rates of CH4 production, but it is unclear whether or not this effect occurs at the ecosystem scale. The goal of this research was to explore the link between algal-derived carbon and methane production in the sediment of a eutrophic reservoir located in southwest Ohio, using a sampling design that capitalized on spatial and temporal gradients in autochthonous carbon input to sediments. Specifically, we aimed to determine if the within-reservoir gradient of sediment algal-derived organic matter and sediment CH4 production rates correlate. This was done by retrieving sediment cores from 15 sites within the reservoir along a known gradient of methane emission rates, at two separate time points in 2016: late spring before the sediments had received large amounts of algal input and mid-summer after algal blooms had been prevalent in the reservoir. Potential CH4 production rates, sediment organic matter source, and microbial community composition were charac

Observations on gas exchange and element recycle within a gas-closed algal-mouse system

NASA Technical Reports Server (NTRS)

Smernoff, D. T.; Wharton, R. A., Jr.; Averner, M. M.

1986-01-01

Life support systems based on bioregeneration rely on the control and manipulation of organisms. Algae are potentially useful for a variety of Closed Ecological Life Support System (CELSS) functions including the revitalization of atmospheres, production of food and for nitrogen fixation. The results of experiments conducted with a gas-closed algal-mouse system designed to investigate gas exchange phenomena under varying algal environmental conditions, and the ability of algae to utilize oxidized mouse solid waste are reported. Inherent instabilities exist between the uptake and release of carbon dioxide (CO2) and oxygen (O2) by the mouse and algae in a gas-closed system. Variations in light intensity and cell density alter the photosynthetic rate of the algae and enable short-term steady-state concentrations of atmospheric CO2 and O2. Different nitrogen sources (urea and nitrate) result in different algal assimilatory quotients (AQ). Combinations of photosynthetic rate and AQ ratio manipulations were examined for their potential in stabilizing atmospheric gas concentrations in the gas-closed algal-mouse system.

Enhancing biomass energy yield from pilot-scale high rate algal ponds with recycling.

PubMed

Park, J B K; Craggs, R J; Shilton, A N

2013-09-01

This paper investigates the effect of recycling on biomass energy yield in High Rate Algal Ponds (HRAPs). Two 8 m(3) pilot-scale HRAPs treating primary settled sewage were operated in parallel and monitored over a 2-year period. Volatile suspended solids were measured from both HRAPs and their gravity settlers to determine biomass productivity and harvest efficiency. The energy content of the biomass was also measured. Multiplying biomass productivity and harvest efficiency gives the 'harvestable biomass productivity' and multiplying this by the energy content defines the actual 'biomass energy yield'. In Year 1, algal recycling was implemented in one of the ponds (HRAPr) and improved harvestable biomass productivity by 58% compared with the control (HRAPc) without recycling (HRAPr: 9.2 g/m(2)/d; HRAPc: 5.8 g/m(2)/d). The energy content of the biomass grown in HRAPr, which was dominated by Pediastrun boryanum, was 25% higher than the control HRAPc which contained a mixed culture of 4-5 different algae (HRAPr: 21.5 kJ/g; HRAPc: 18.6 kJ/g). In Year 2, HRAPc was then seeded with the biomass harvested from the P. boryanum dominated HRAPr. This had the effect of shifting algal dominance from 89% Dictyosphaerium sp. (which is poorly-settleable) to over 90% P. boryanum in 5 months. Operation of this pond was then switched to recycling its own harvested biomass, which maintained P. boryanum dominance for the rest of Year 2. This result confirms, for the first time in the literature, that species control is possible for similarly sized co-occurring algal colonies in outdoor HRAP by algal recycling. With regard to the overall improvement in biomass energy yield, which is a critical parameter in the context of algal cultivation for biofuels, the combined improvements that recycling triggered in biomass productivity, harvest efficiency and energy content enhanced the harvested biomass energy yield by 66% (HRAPr: 195 kJ/m(2)/day; HRAPc: 118 kJ/m(2)/day). Copyright © 2013

A 3D cell culture system: separation distance between INS-1 cell and endothelial cell monolayers co-cultured in fibrin influences INS-1 cells insulin secretion.

PubMed

Sabra, Georges; Vermette, Patrick

2013-02-01

The aim of this study was to develop an in vitro cell culture system allowing studying the effect of separation distance between monolayers of rat insulinoma cells (INS-1) and human umbilical vein endothelial cells (HUVEC) co-cultured in fibrin over INS-1 cell insulin secretion. For this purpose, a three-dimensional (3D) cell culture chamber was designed, built using micro-fabrication techniques and validated. The co-culture was successfully carried out and the effect on INS-1 cell insulin secretion was investigated. After 48 and 72 h, INS-1 cells co-cultured with HUVEC separated by a distance of 100 µm revealed enhanced insulin secretion compared to INS-1 cells cultured alone or co-cultured with HUVEC monolayers separated by a distance of 200 µm. These results illustrate the importance of the separation distance between two cell niches for cell culture design and the possibility to further enhance the endocrine function of beta cells when this factor is considered. Copyright © 2012 Wiley Periodicals, Inc.

Cell Culturing of Cytoskeleton

NASA Technical Reports Server (NTRS)

2004-01-01

Biomedical research offers hope for a variety of medical problems, from diabetes to the replacement of damaged bone and tissues. Bioreactors, which are used to grow cells and tissue cultures, play a major role in such research and production efforts. Cell culturing, such as this bone cell culture, is an important part of biomedical research. The BioDyn payload includes a tissue engineering investigation. The commercial affiliate, Millenium Biologix, Inc., has been conducting bone implant experiments to better understand how synthetic bone can be used to treat bone-related illnesses and bone damaged in accidents. On STS-95, the BioDyn payload will include a bone cell culture aimed to help develop this commercial synthetic bone product. Millenium Biologix, Inc., is exploring the potential for making human bone implantable materials by seeding its proprietary artificial scaffold material with human bone cells. The product of this tissue engineering experiment using the Bioprocessing Modules (BPMs) on STS-95 is space-grown bone implants, which could have potential for dental implants, long bone grafts, and coating for orthopedic implants such as hip replacements.

Cell Culturing of Cytoskeleton

NASA Technical Reports Server (NTRS)

2004-01-01

Biomedical research offers hope for a variety of medical problems, from diabetes to the replacement of damaged bone and tissues. Bioreactors, which are used to grow cells and tissue cultures, play a major role in such research and production efforts. Cell culturing, such as this bone cell culture, is an important part of biomedical research. The BioDyn payload includes a tissue engineering investigation. The commercial affiliate, Millenium Biologix, Inc. has been conducting bone implant experiments to better understand how synthetic bone can be used to treat bone-related illnesses and bone damaged in accidents. On STS-95, the BioDyn payload will include a bone cell culture aimed to help develop this commercial synthetic bone product. Millenium Biologix, Inc. is exploring the potential for making human bone implantable materials by seeding its proprietary artificial scaffold material with human bone cells. The product of this tissue engineering experiment using the Bioprocessing Modules (BPMs) on STS-95 is space-grown bone implants, which could have potential for dental implants, long bone grafts, and coating for orthopedic implants such as hip replacements.

9 CFR 101.6 - Cell cultures.

Code of Federal Regulations, 2012 CFR

2012-01-01

... 9 Animals and Animal Products 1 2012-01-01 2012-01-01 false Cell cultures. 101.6 Section 101.6..., SERUMS, TOXINS, AND ANALOGOUS PRODUCTS; ORGANISMS AND VECTORS DEFINITIONS § 101.6 Cell cultures. When used in conjunction with or in reference to cell cultures, which may be referred to as tissue cultures...

9 CFR 101.6 - Cell cultures.

Code of Federal Regulations, 2013 CFR

2013-01-01

... 9 Animals and Animal Products 1 2013-01-01 2013-01-01 false Cell cultures. 101.6 Section 101.6..., SERUMS, TOXINS, AND ANALOGOUS PRODUCTS; ORGANISMS AND VECTORS DEFINITIONS § 101.6 Cell cultures. When used in conjunction with or in reference to cell cultures, which may be referred to as tissue cultures...

9 CFR 101.6 - Cell cultures.

Code of Federal Regulations, 2014 CFR

2014-01-01

... 9 Animals and Animal Products 1 2014-01-01 2014-01-01 false Cell cultures. 101.6 Section 101.6..., SERUMS, TOXINS, AND ANALOGOUS PRODUCTS; ORGANISMS AND VECTORS DEFINITIONS § 101.6 Cell cultures. When used in conjunction with or in reference to cell cultures, which may be referred to as tissue cultures...

9 CFR 101.6 - Cell cultures.

Code of Federal Regulations, 2011 CFR

2011-01-01

... 9 Animals and Animal Products 1 2011-01-01 2011-01-01 false Cell cultures. 101.6 Section 101.6..., SERUMS, TOXINS, AND ANALOGOUS PRODUCTS; ORGANISMS AND VECTORS DEFINITIONS § 101.6 Cell cultures. When used in conjunction with or in reference to cell cultures, which may be referred to as tissue cultures...

9 CFR 101.6 - Cell cultures.

Code of Federal Regulations, 2010 CFR

2010-01-01

... 9 Animals and Animal Products 1 2010-01-01 2010-01-01 false Cell cultures. 101.6 Section 101.6..., SERUMS, TOXINS, AND ANALOGOUS PRODUCTS; ORGANISMS AND VECTORS DEFINITIONS § 101.6 Cell cultures. When used in conjunction with or in reference to cell cultures, which may be referred to as tissue cultures...

[Growth and biochemical composition of thalassiosira pseudonana (Thalassiosirales: Thalassiosiraceae) cultivated in semicontinuous system at different culture media and irradiances].

PubMed

Vásquez-Suárez, Aleikar; Guevara, Miguel; González, Mayelys; Cortez, Roraysi; Arredondo-Vega, Bertha

2013-09-01

Thalassiosira pseudonana is a marine Bacillariophyta commonly used as live feed in mariculture. The growth rate and biochemical composition of microalgae are highly influenced by environmental factors such as, irradiance and nutrient availability. The aim of this study was to investigate the influence of three irradiances (60, 120 and 180 microE/m2.s) and two culture media (Algal and Humus) on growth and biochemical composition of this diatom. The microalga was grown semicontinuously at a daily renewal rate of fresh media of 30%, 37 per thousand salinity, 25 +/- 1 degree C and constant aeration (200 mL/min). The cell densities (cel/mL) and contents of protein, lipid, carbohydrate, chlorophyll a, total carotenoids, and fatty acids, showed significant differences (p < 0.05) between treatments. During steady-state phase, the maximal cell density, and lipid and carbohydrate contents were of 4.62 x 10(6) cel/mL, 20.3 +/- 2.28% and 16.6 +/- 2.43%, respectively, and were achieved in Humus medium at 180 microE/ m2.s. Moreover, highest protein contents (45.0 +/- 5.05%) and total carotenoids (0.5 +/- 0.01%) were obtained in Algal medium at 180 microE/m2.s. Chlorophyll a (0.93 +/- 0.04%) was higher at low irradiances in Algal medium. In both media, the fatty acids unsaturation degree was lower with increasing irradiance, being eicosapentaenoic acid, 20:5 n-3 (EPA) most represented (6.20%) in Algal medium at 60 microE/m2.s. This strain of T. pseudonana showed multiple physiological responses to changes in culture conditions, and may be cultivated with an alternative medium, which reduced the operating costs and allowed a high nutritional biomass production value for animals under culture.

The ins and outs of algal metal transport

PubMed Central

Blaby-Haas, Crysten E.; Merchant, Sabeeha S.

2012-01-01

Metal transporters are a central component in the interaction of algae with their environment. They represent the first line of defense to cellular perturbations in metal concentration, and by analyzing algal metal transporter repertoires, we gain insight into a fundamental aspect of algal biology. The ability of individual algae to thrive in environments with unique geochemistry, compared to non-algal species commonly used as reference organisms for metal homeostasis, provides an opportunity to broaden our understanding of biological metal requirements, preferences and trafficking. Chlamydomonas reinhardtii is the best developed reference organism for the study of algal biology, especially with respect to metal metabolism; however, the diversity of algal niches necessitates a comparative genomic analysis of all sequenced algal genomes. A comparison between known and putative proteins in animals, plants, fungi and algae using protein similarity networks has revealed the presence of novel metal metabolism components in Chlamydomonas including new iron and copper transporters. This analysis also supports the concept that, in terms of metal metabolism, algae from similar niches are more related to one another than to algae from the same phylogenetic clade. PMID:22569643

Assessing the potential of polyculture to accelerate algal biofuel production

DOE PAGES

Newby, Deborah T.; Mathews, Teresa J.; Pate, Ron C.; ...

2016-10-24

To date, the algal biofuel industry has focused on the cultivation of monocultures of highly productive algal strains, but scaling up production remains challenging. However, algal monocultures are difficult to maintain because they are easily contaminated by wild algal strains, grazers, and pathogens. In contrast, theory suggests that polycultures (multispecies assemblages) can promote both ecosystem stability and productivity. A greater understanding of species interactions and how communities change with time will need to be developed before polycultures can be successfully applied to large-scale algal production efforts. Here in this paper we review the agricultural and ecological literature to explore opportunitiesmore » for increased annual biomass production through the use of algal polycultures. We discuss case studies where algal polycultures have been successfully maintained for industries other than the biofuel industry, as well as the few studies that have compared biomass production of algal polycultures to that of monocultures. Assemblages that include species with complementary traits are of particular promise. These assemblages have the potential not only to increase crop productivity and stability, but they may also be capable of utilizing natural resources (e.g. light, nutrients, water) more efficiently via tighter niche packing. Therefore, algal polycultures show promise for enhancing biomass productivity, enabling sustainable production and reducing overall production costs.« less

Assessing the potential of polyculture to accelerate algal biofuel production

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

Newby, Deborah T.; Mathews, Teresa J.; Pate, Ron C.

To date, the algal biofuel industry has focused on the cultivation of monocultures of highly productive algal strains, but scaling up production remains challenging. However, algal monocultures are difficult to maintain because they are easily contaminated by wild algal strains, grazers, and pathogens. In contrast, theory suggests that polycultures (multispecies assemblages) can promote both ecosystem stability and productivity. A greater understanding of species interactions and how communities change with time will need to be developed before polycultures can be successfully applied to large-scale algal production efforts. Here in this paper we review the agricultural and ecological literature to explore opportunitiesmore » for increased annual biomass production through the use of algal polycultures. We discuss case studies where algal polycultures have been successfully maintained for industries other than the biofuel industry, as well as the few studies that have compared biomass production of algal polycultures to that of monocultures. Assemblages that include species with complementary traits are of particular promise. These assemblages have the potential not only to increase crop productivity and stability, but they may also be capable of utilizing natural resources (e.g. light, nutrients, water) more efficiently via tighter niche packing. Therefore, algal polycultures show promise for enhancing biomass productivity, enabling sustainable production and reducing overall production costs.« less

Tryptophan oxidation catabolite, N-formylkynurenine, in photo degraded cell culture medium results in reduced cell culture performance.

PubMed

McElearney, Kyle; Ali, Amr; Gilbert, Alan; Kshirsagar, Rashmi; Zang, Li

2016-01-01

Chemically defined media have been widely used in the biopharmaceutical industry to enhance cell culture productivities and ensure process robustness. These media, which are quite complex, often contain a mixture of many components such as vitamins, amino acids, metals and other chemicals. Some of these components are known to be sensitive to various stress factors including photodegradation. Previous work has shown that small changes in impurity concentrations induced by these potential stresses can have a large impact on the cell culture process including growth and product quality attributes. Furthermore, it has been shown to be difficult to detect these modifications analytically due to the complexity of the cell culture media and the trace level of the degradant products. Here, we describe work performed to identify the specific chemical(s) in photodegraded medium that affect cell culture performance. First, we developed a model system capable of detecting changes in cell culture performance. Second, we used these data and applied an LC-MS analytical technique to characterize the cell culture media and identify degradant products which affect cell culture performance. Riboflavin limitation and N-formylkynurenine (NFK), a tryptophan oxidation catabolite, were identified as chemicals which results in a reduction in cell culture performance. © 2015 American Institute of Chemical Engineers.

Sustainable Algal Energy Production and Environmental Remediation

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

Cooke, William E.

2012-07-14

Overall, our results confirm that wild algal species sequester a wide range of organic and metal contaminants and excess nutrients (PAHs, trace metals, and nutrients) from natural waters, and suggest parameters that could be useful in predicting uptake rates for algae growing on an algal floway or other algal growth systems in the environment or in industrial processes. The implication for various fuel production processes differ with the detailed unit operations involved, and these results will be of use in the developing of scaling experiments for various types of engineering process designs.

Producing docosahexaenoic acid (DHA)-rich algae from biodiesel-derived crude glycerol: effects of impurities on DHA production and algal biomass composition.

PubMed

Pyle, Denver J; Garcia, Rafael A; Wen, Zhiyou

2008-06-11

Crude glycerol is the primary byproduct of the biodiesel industry. Producing docosahexaenoic acid (DHA, 22:6 n-3) through fermentation of the alga Schizochytrium limacinum on crude glycerol provides a unique opportunity to utilize a large quantity of this byproduct. The objective of this work is to investigate the effects of impurities contained in the crude glycerol on DHA production and algal biomass composition. Crude glycerol streams were obtained from different biodiesel refineries. All of the glycerol samples contained methanol, soaps, and various elements including calcium, phosphorus, potassium, silicon, sodium, and zinc. Both methanol and soap were found to negatively influence algal DHA production; these two impurities can be removed from culture medium by evaporation through autoclaving (for methanol) and by precipitation through pH adjustment (for soap). The glycerol-derived algal biomass contained 45-50% lipid, 14-20% protein, and 25% carbohydrate, with 8-13% ash content. Palmitic acid (C16:0) and DHA were the two major fatty acids in the algal lipid. The algal biomass was rich in lysine and cysteine, relative to many common feedstuffs. Elemental analysis by inductively coupled plasma showed that boron, calcium, copper, iron, magnesium, phosphorus, potassium, silicon, sodium, and sulfur were present in the biomass, whereas no heavy metals (such as mercury) were detected in the algal biomass. Overall, the results show that crude glycerol was a suitable carbon source for algal fermentation. The crude glycerol-derived algal biomass had a high level of DHA and a nutritional profile similar to that of commercial algal biomass, suggesting a great potential for using crude glycerol-derived algae in omega-3-fortified food or feed.

Algal biodiesel economy and competition among bio-fuels.

PubMed

Lee, D H

2011-01-01

This investigation examines the possible results of policy support in developed and developing economies for developing algal biodiesel through to 2040. This investigation adopts the Taiwan General Equilibrium Model-Energy for Bio-fuels (TAIGEM-EB) to predict competition among the development of algal biodiesel, bioethanol and conventional crop-based biodiesel. Analytical results show that algal biodiesel will not be the major energy source in 2040 without strong support in developed economies. In contrast, bioethanol enjoys a development advantage relative to both forms of biodiesel. Finally, algal biodiesel will almost completely replace conventional biodiesel. CO(2) reduction benefits the development of the bio-fuels industry. Copyright © 2010 Elsevier Ltd. All rights reserved.

Proterozoic microfossils revealing the time of algal divergences

NASA Astrophysics Data System (ADS)

Moczydlowska-Vidal, Malgorzata

2010-05-01

Proterozoic microfossils revealing the time of algal divergences Małgorzata Moczydłowska-Vidal Uppsala University, Department of Earth Sciences, Palaeobiology, Villavägen 16, SE 752 36 Uppsala, Sweden (malgo.vidal@pal.uu.se) Morphological and reproductive features and cell wall ultrastructure and biochemistry of Proterozoic acritarchs are used to determine their affinity to modern algae. The first appearance datum of these microbiota is traced to infer a minimum age of the divergence of the algal classes to which they may belong. The chronological appearance of microfossils that represent phycoma-like and zygotic cysts and vegetative cells and/or aplanospores, respectively interpreted as prasinophyceaen and chlorophyceaen microalgae, is related to the Viridiplantae phylogeny. These divergence times differ from molecular clock estimates, and the palaeontological evidence suggests that they are older. The best examples of unicellular, organic-walled microfossils (acritarchs) from the Mesoproterozoic to Early Ordovician are reviewed to demonstrate features, which are indicative of their affinity to photosynthetic microalgae. The first indication that a microfossil may be algal is a decay- and acid-resistant cell wall, which reflects its biochemistry and ultrastructure, and probably indicates the ability to protect a resting/reproductive cyst. The biopolymers synthesized in the cell walls of algae and in land plants ("plant cells"), such as sporopollenin/algaenan, are diagnostic for photosynthetic taxa and were inherited from early unicellular ancestors. These preservable cell walls are resistant to acetolysis, hydrolysis and acids, and show diagnostic ultrastructures such as the trilaminar sheath structure (TLS). "Plant cell" walls differ in terms of chemical compounds, which give high preservation potential, from fungal and animal cell walls. Fungal and animal cells are fossilized only by syngenetic permineralization, whereas "plant cells" are fossilized as body

Algal genes in the closest relatives of animals.

PubMed

Sun, Guiling; Yang, Zefeng; Ishwar, Arjun; Huang, Jinling

2010-12-01

The spread of photosynthesis is one of the most important but controversial topics in eukaryotic evolution. Because of massive gene transfer from plastids to the nucleus and because of the possibility that plastids have been lost in evolution, algal genes in aplastidic organisms often are interpreted as footprints of photosynthetic ancestors. These putative plastid losses, in turn, have been cited as support for scenarios involving the spread of plastids in broadscale eukaryotic evolution. Phylogenomic analyses identified more than 100 genes of possible algal origin in Monosiga, a unicellular species from choanoflagellates, a group considered to be the closest protozoan relatives of animals and to be primitively heterotrophic. The vast majority of these algal genes appear to be derived from haptophytes, diatoms, or green plants. Furthermore, more than 25% of these algal genes are ultimately of prokaryotic origin and were spread secondarily to Monosiga. Our results show that the presence of algal genes may be expected in many phagotrophs or taxa of phagotrophic ancestry and therefore does not necessarily represent evidence of plastid losses. The ultimate prokaryotic origin of some algal genes and their simultaneous presence in both primary and secondary photosynthetic eukaryotes either suggest recurrent gene transfer events under specific environments or support a more ancient origin of primary plastids.

Evidence for water-mediated mechanisms in coral–algal interactions

PubMed Central

Jorissen, Hendrikje; Skinner, Christina; Osinga, Ronald; de Beer, Dirk

2016-01-01

Although many coral reefs have shifted from coral-to-algal dominance, the consequence of such a transition for coral–algal interactions and their underlying mechanisms remain poorly understood. At the microscale, it is unclear how diffusive boundary layers (DBLs) and surface oxygen concentrations at the coral–algal interface vary with algal competitors and competitiveness. Using field observations and microsensor measurements in a flow chamber, we show that coral (massive Porites) interfaces with thick turf algae, macroalgae, and cyanobacteria, which are successful competitors against coral in the field, are characterized by a thick DBL and hypoxia at night. In contrast, coral interfaces with crustose coralline algae, conspecifics, and thin turf algae, which are poorer competitors, have a thin DBL and low hypoxia at night. Furthermore, DBL thickness and hypoxia at the interface with turf decreased with increasing flow speed, but not when thick turf was upstream. Our results support the importance of water-mediated transport mechanisms in coral–algal interactions. Shifts towards algal dominance, particularly dense assemblages, may lead to thicker DBLs, higher hypoxia, and higher concentrations of harmful metabolites and pathogens along coral borders, which in turn may facilitate algal overgrowth of live corals. These effects may be mediated by flow speed and orientation. PMID:27512146

Evidence for water-mediated mechanisms in coral-algal interactions.

PubMed

Jorissen, Hendrikje; Skinner, Christina; Osinga, Ronald; de Beer, Dirk; Nugues, Maggy M

2016-08-17

Although many coral reefs have shifted from coral-to-algal dominance, the consequence of such a transition for coral-algal interactions and their underlying mechanisms remain poorly understood. At the microscale, it is unclear how diffusive boundary layers (DBLs) and surface oxygen concentrations at the coral-algal interface vary with algal competitors and competitiveness. Using field observations and microsensor measurements in a flow chamber, we show that coral (massive Porites) interfaces with thick turf algae, macroalgae, and cyanobacteria, which are successful competitors against coral in the field, are characterized by a thick DBL and hypoxia at night. In contrast, coral interfaces with crustose coralline algae, conspecifics, and thin turf algae, which are poorer competitors, have a thin DBL and low hypoxia at night. Furthermore, DBL thickness and hypoxia at the interface with turf decreased with increasing flow speed, but not when thick turf was upstream. Our results support the importance of water-mediated transport mechanisms in coral-algal interactions. Shifts towards algal dominance, particularly dense assemblages, may lead to thicker DBLs, higher hypoxia, and higher concentrations of harmful metabolites and pathogens along coral borders, which in turn may facilitate algal overgrowth of live corals. These effects may be mediated by flow speed and orientation. © 2016 The Author(s).

Algal turf scrubber (ATS) floways on the Great Wicomico River, Chesapeake Bay: productivity, algal community structure, substrate and chemistry(1).

PubMed

Adey, Walter H; Laughinghouse, H Dail; Miller, John B; Hayek, Lee-Ann C; Thompson, Jesse G; Bertman, Steven; Hampel, Kristin; Puvanendran, Shanmugam

2013-06-01

Two Algal Turf Scrubber (ATS) units were deployed on the Great Wicomico River (GWR) for 22 months to examine the role of substrate in increasing algal productivity and nutrient removal. The yearly mean productivity of flat ATS screens was 15.4 g · m(-2)  · d(-1) . This was elevated to 39.6 g · m(-2)  · d(-1) with a three-dimensional (3-D) screen, and to 47.7 g · m(-2)  · d(-1) by avoiding high summer harvest temperatures. These methods enhanced nutrient removal (N, P) in algal biomass by 3.5 times. Eighty-six algal taxa (Ochrophyta [diatoms], Chlorophyta [green algae], and Cyan-obacteria [blue-green algae]) self-seeded from the GWR and demonstrated yearly cycling. Silica (SiO2 ) content of the algal biomass ranged from 30% to 50% of total biomass; phosphorus, nitrogen, and carbon content of the total algal biomass ranged from 0.15% to 0.21%, 2.13% to 2.89%, and 20.0% to 25.7%, respectively. Carbohydrate content (at 10%-25% of AFDM) was dominated by glucose. Lipids (fatty acid methyl ester; FAMEs) ranged widely from 0.5% to 9% AFDM, with Omega-3 fatty acids a consistent component. Mathematical modeling of algal produ-ctivity as a function of temperature, light, and substrate showed a proportionality of 4:3:3, resp-ectively. Under landscape ATS operation, substrate manipulation provides a considerable opportunity to increase ATS productivity, water quality amelioration, and biomass coproduction for fertilizers, fermentation energy, and omega-3 products. Based on the 3-D prod-uctivity and algal chemical composition demonstrated, ATS systems used for nonpoint source water treat-ment can produce ethanol (butanol) at 5.8× per unit area of corn, and biodiesel at 12.0× per unit area of soy beans (agricultural production US). © 2013 Phycological Society of America.

Outdoor cultures of Chlorella pyrenoidosa in the effluent of anaerobically digested activated sludge: The effects of pH and free ammonia.

PubMed

Tan, Xiao-Bo; Zhang, Ya-Lei; Yang, Li-Bin; Chu, Hua-Qiang; Guo, Jun

2016-01-01

A freshwater algae Chlorella pyrenoidosa was cultured outdoors using anaerobically digested activated sludge effluent. The effects of pH variations were evaluated. The coupled pH variations and free ammonia toxicity significantly affected the algal growth, lipids accumulation and contamination control during every season. The free ammonia toxicity at high pH levels actually inhibited the algal growth. Compared to an optimal algal growth at a pH of 5.7-6.5, biomass productivity at a high pH of 8.3-8.8 was reduced by 67.15±6.98%, 54.39±6.42% and 83.63±5.71% in the spring, fall and summer, respectively. When the pH rose above 9.1-9.6, algae were unable to grow in the wastewater. However, high pH levels reduced contamination (e.g., bacteria and microalgae grazers) and triggered lipids accumulation in algal cells. These findings suggest that pH control strategies are essential for this type of algal wastewater system, where ammonia is the dominant nitrogen source. Copyright © 2015 Elsevier Ltd. All rights reserved.

Microfluidic cell culture systems for drug research.

PubMed

Wu, Min-Hsien; Huang, Song-Bin; Lee, Gwo-Bin

2010-04-21

In pharmaceutical research, an adequate cell-based assay scheme to efficiently screen and to validate potential drug candidates in the initial stage of drug discovery is crucial. In order to better predict the clinical response to drug compounds, a cell culture model that is faithful to in vivo behavior is required. With the recent advances in microfluidic technology, the utilization of a microfluidic-based cell culture has several advantages, making it a promising alternative to the conventional cell culture methods. This review starts with a comprehensive discussion on the general process for drug discovery and development, the role of cell culture in drug research, and the characteristics of the cell culture formats commonly used in current microfluidic-based, cell-culture practices. Due to the significant differences in several physical phenomena between microscale and macroscale devices, microfluidic technology provides unique functionality, which is not previously possible by using traditional techniques. In a subsequent section, the niches for using microfluidic-based cell culture systems for drug research are discussed. Moreover, some critical issues such as cell immobilization, medium pumping or gradient generation in microfluidic-based, cell-culture systems are also reviewed. Finally, some practical applications of microfluidic-based, cell-culture systems in drug research particularly those pertaining to drug toxicity testing and those with a high-throughput capability are highlighted.

Algal culture studies related to a closed ecological life support system

NASA Technical Reports Server (NTRS)

Radmer, R.; Behrens, P.; Fernandez, E.; Ollinger, O.; Howell, C.

1984-01-01

Studies on the steady-state long-term (4 month) culture of Scenedesmus obliquus algae, maintained in an annular air-lift column operated as a turbidostat, were carried out to evaluate the life-supporting possibilities of this system. Chlorophyll production and cell number as functions of the dry weight were linear at constant illumination. Productivity (measured as the product of dry weight, mg/ml, and the growth rate, ml/hr) vs. dry weight rose linearly until the cell density reached a level at which light became limiting (89 percent absorption of the photosynthetically active radiation). In the initial, linear portion of the curve, the productivity was limited by cell growth at the given light intensity. The maximum dilution rate of the system corresponded to the doubling time of 13.4 hr, about half the maximum rate, with a productivity of 80 percent of the maximum theoretical productivity. The high light utilization efficiencies were contributed by the low (10 percent of full sunlight) incident intensities.

Tocopherol production in plant cell cultures.

PubMed

Caretto, Sofia; Nisi, Rossella; Paradiso, Annalisa; De Gara, Laura

2010-05-01

Tocopherols, collectively known as vitamin E, are lipophilic antioxidants, essential dietary components for mammals and exclusively synthesized by photosynthetic organisms. Of the four forms (alpha, beta, gamma and delta), alpha-tocopherol is the major vitamin E form present in green plant tissues, and has the highest vitamin E activity. Synthetic alpha-tocopherol, being a racemic mixture of eight different stereoisomers, always results less effective than the natural form (R,R,R) alpha-tocopherol. This raises interest in obtaining this molecule from natural sources, such as plant cell cultures. Plant cell and tissue cultures are able to produce and accumulate valuable metabolites that can be used as food additives, nutraceuticals and pharmaceuticals. Sunflower cell cultures, growing under heterotrophic conditions, were exploited to establish a suitable in vitro production system of natural alpha-tocopherol. Optimization of culture conditions, precursor feeding and elicitor application were used to improve the tocopherol yields of these cultures. Furthermore, these cell cultures were useful to investigate the relationship between alpha-tocopherol biosynthesis and photomixotrophic culture conditions, revealing the possibility to enhance tocopherol production by favouring sunflower cell photosynthetic properties. The modulation of alpha-tocopherol levels in plant cell cultures can provide useful hints for a regulatory impact on tocopherol metabolism.

Use of an adaptable cell culture kit for performing lymphocyte and monocyte cell cultures in microgravity

NASA Technical Reports Server (NTRS)

Hatton, J. P.; Lewis, M. L.; Roquefeuil, S. B.; Chaput, D.; Cazenave, J. P.; Schmitt, D. A.

1998-01-01

The results of experiments performed in recent years on board facilities such as the Space Shuttle/Spacelab have demonstrated that many cell systems, ranging from simple bacteria to mammalian cells, are sensitive to the microgravity environment, suggesting gravity affects fundamental cellular processes. However, performing well-controlled experiments aboard spacecraft offers unique challenges to the cell biologist. Although systems such as the European 'Biorack' provide generic experiment facilities including an incubator, on-board 1-g reference centrifuge, and contained area for manipulations, the experimenter must still establish a system for performing cell culture experiments that is compatible with the constraints of spaceflight. Two different cell culture kits developed by the French Space Agency, CNES, were recently used to perform a series of experiments during four flights of the 'Biorack' facility aboard the Space Shuttle. The first unit, Generic Cell Activation Kit 1 (GCAK-1), contains six separate culture units per cassette, each consisting of a culture chamber, activator chamber, filtration system (permitting separation of cells from supernatant in-flight), injection port, and supernatant collection chamber. The second unit (GCAK-2) also contains six separate culture units, including a culture, activator, and fixation chambers. Both hardware units permit relatively complex cell culture manipulations without extensive use of spacecraft resources (crew time, volume, mass, power), or the need for excessive safety measures. Possible operations include stimulation of cultures with activators, separation of cells from supernatant, fixation/lysis, manipulation of radiolabelled reagents, and medium exchange. Investigations performed aboard the Space Shuttle in six different experiments used Jurkat, purified T-cells or U937 cells, the results of which are reported separately. We report here the behaviour of Jurkat and U937 cells in the GCAK hardware in ground

Algal Toxins Alter Copepod Feeding Behavior

PubMed Central

Hong, Jiarong; Talapatra, Siddharth; Katz, Joseph; Tester, Patricia A.; Waggett, Rebecca J.; Place, Allen R.

2012-01-01

Using digital holographic cinematography, we quantify and compare the feeding behavior of free-swimming copepods, Acartia tonsa, on nutritional prey (Storeatula major) to that occurring during exposure to toxic and non-toxic strains of Karenia brevis and Karlodinium veneficum. These two harmful algal species produce polyketide toxins with different modes of action and potency. We distinguish between two different beating modes of the copepod’s feeding appendages–a “sampling beating” that has short durations (<100 ms) and involves little fluid entrainment and a longer duration “grazing beating” that persists up to 1200 ms and generates feeding currents. The durations of both beating modes have log-normal distributions. Without prey, A. tonsa only samples the environment at low frequency. Upon introduction of non-toxic food, it increases its sampling time moderately and the grazing period substantially. On mono algal diets for either of the toxic dinoflagellates, sampling time fraction is high but the grazing is very limited. A. tonsa demonstrates aversion to both toxic algal species. In mixtures of S. major and the neurotoxin producing K. brevis, sampling and grazing diminish rapidly, presumably due to neurological effects of consuming brevetoxins while trying to feed on S. major. In contrast, on mixtures of cytotoxin producing K. veneficum, both behavioral modes persist, indicating that intake of karlotoxins does not immediately inhibit the copepod’s grazing behavior. These findings add critical insight into how these algal toxins may influence the copepod’s feeding behavior, and suggest how some harmful algal species may alter top-down control exerted by grazers like copepods. PMID:22629336

Eukaryotic algal phytochromes span the visible spectrum

PubMed Central

Rockwell, Nathan C.; Duanmu, Deqiang; Martin, Shelley S.; Bachy, Charles; Price, Dana C.; Bhattacharya, Debashish; Worden, Alexandra Z.; Lagarias, J. Clark

2014-01-01

Plant phytochromes are photoswitchable red/far-red photoreceptors that allow competition with neighboring plants for photosynthetically active red light. In aquatic environments, red and far-red light are rapidly attenuated with depth; therefore, photosynthetic species must use shorter wavelengths of light. Nevertheless, phytochrome-related proteins are found in recently sequenced genomes of many eukaryotic algae from aquatic environments. We examined the photosensory properties of seven phytochromes from diverse algae: four prasinophyte (green algal) species, the heterokont (brown algal) Ectocarpus siliculosus, and two glaucophyte species. We demonstrate that algal phytochromes are not limited to red and far-red responses. Instead, different algal phytochromes can sense orange, green, and even blue light. Characterization of these previously undescribed photosensors using CD spectroscopy supports a structurally heterogeneous chromophore in the far-red–absorbing photostate. Our study thus demonstrates that extensive spectral tuning of phytochromes has evolved in phylogenetically distinct lineages of aquatic photosynthetic eukaryotes. PMID:24567382

Repeated batch cultivation of the hydrocarbon-degrading, micro-algal strain Prototheca zopfii RND16 immobilized in polyurethane foam.

PubMed

Ueno, Ryohei; Wada, Shun; Urano, Naoto

2008-01-01

This study reports on the stability of the cells of a heterotrophic green micro-algal strain Prototheca zopfii RND16 immobilized in polyurethane foam (PUF) cubes during degradation of mixed hydrocarbon substrate, which was composed of n-alkanes and polycyclic aromatic hydrocarbons (PAHs), in 5 successive cycles of repeated batch cultivation at 30 degrees C. Both RND16 cells and mixed hydrocarbon substrate components had been entrapped in PUF cubes through cultivation. PUF-immobilized RND16 degraded n-alkanes almost completely, whereas the strain hardly degraded PAHs in PUFs, rather they accumulated in the matrices. It is noteworthy that this result is strikingly different from that of the free-living cell culture, where RND16 reduced concentrations of both n-alkanes and PAHs. However, PAHs accumulation in the PUFs did not impair the performance of the immobilized alga to utilize n-alkanes. These results suggest that the PUFs harboring RND16 cells could be used repeatedly for selective retrieval of PAHs from oil-polluted waters after preferential biodegradation of n-alkanes by algae.

Exploiting algal NADPH oxidase for biophotovoltaic energy

DOE PAGES

Anderson, Alexander; Laohavisit, Anuphon; Blaby, Ian K.; ...

2015-01-29

Photosynthetic microbes exhibit light-dependent electron export across the cell membrane, which can generate electricity in biological photovoltaic (BPV) devices. How electrons are exported remains to be determined; the identification of mechanisms would help selection or generation of photosynthetic microbes capable of enhanced electrical output. We show that plasma membrane NADPH oxidase activity is a significant component of light-dependent generation of electricity by the unicellular green alga Chlamydomonas reinhardtii. NADPH oxidases export electrons across the plasma membrane to form superoxide anion from oxygen. The C. reinhardtii mutant lacking the NADPH oxidase encoded by RBO1 is impaired in both extracellular superoxide anionmore » production and current generation in a BPV device. Complementation with the wild-type gene restores both capacities, demonstrating the role of the enzyme in electron export. Monitoring light-dependent extracellular superoxide production with a colorimetric assay is shown to be an effective way of screening for electrogenic potential of candidate algal strains. Furthermore, the results show that algal NADPH oxidases are important for superoxide anion production and open avenues for optimizing the biological component of these devices.« less

TEXAS HARMFUL ALGAL BLOOM COORDINATION MX964014

EPA Science Inventory

Harmful algal blooms (HAB) are an expanding problem in coastal Texas. Nearly � of the known harmful algal blooms along the Texas coast have occurred in the past ten years and have led to significant resource and tourism losses. For example, there are at least two types of toxic...

Life cycle environmental impacts of wastewater-based algal biofuels.

PubMed

Mu, Dongyan; Min, Min; Krohn, Brian; Mullins, Kimberley A; Ruan, Roger; Hill, Jason

2014-10-07

Recent research has proposed integrating wastewater treatment with algae cultivation as a way of producing algal biofuels at a commercial scale more sustainably. This study evaluates the environmental performance of wastewater-based algal biofuels with a well-to-wheel life cycle assessment (LCA). Production pathways examined include different nutrient sources (municipal wastewater influent to the activated sludge process, centrate from the sludge drying process, swine manure, and freshwater with synthetic fertilizers) combined with emerging biomass conversion technologies (microwave pyrolysis, combustion, wet lipid extraction, and hydrothermal liquefaction). Results show that the environmental performance of wastewater-based algal biofuels is generally better than freshwater-based algal biofuels, but depends on the characteristics of the wastewater and the conversion technologies. Of 16 pathways compared, only the centrate cultivation with wet lipid extraction pathway and the centrate cultivation with combustion pathway have lower impacts than petroleum diesel in all environmental categories examined (fossil fuel use, greenhouse gas emissions, eutrophication potential, and consumptive water use). The potential for large-scale implementation of centrate-based algal biofuel, however, is limited by availability of centrate. Thus, it is unlikely that algal biofuels can provide a large-scale and environmentally preferable alternative to petroleum transportation fuels without considerable improvement in current production technologies. Additionally, the cobenefit of wastewater-based algal biofuel production as an alternate means of treating various wastewaters should be further explored.

Dissolved organic matter reduces algal accumulation of methylmercury

USGS Publications Warehouse

Luengen, Allison C.; Fisher, Nicholas S.; Bergamaschi, Brian A.

2012-01-01

Dissolved organic matter (DOM) significantly decreased accumulation of methylmercury (MeHg) by the diatom Cyclotella meneghiniana in laboratory experiments. Live diatom cells accumulated two to four times more MeHg than dead cells, indicating that accumulation may be partially an energy-requiring process. Methylmercury enrichment in diatoms relative to ambient water was measured by a volume concentration factor (VCF). Without added DOM, the maximum VCF was 32 x 104, and the average VCF (from 10 to 72 h) over all experiments was 12.6 x 104. At very low (1.5 mg/L) added DOM, VCFs dropped by approximately half. At very high (20 mg/L) added DOM, VCFs dropped 10-fold. Presumably, MeHg was bound to a variety of reduced sulfur sites on the DOM, making it unavailable for uptake. Diatoms accumulated significantly more MeHg when exposed to transphilic DOM extracts than hydrophobic ones. However, algal lysate, a labile type of DOM created by resuspending a marine diatom in freshwater, behaved similarly to a refractory DOM isolate from San Francisco Bay. Addition of 67 μM L-cysteine resulted in the largest drop in VCFs, to 0.28 x 104. Although the DOM composition influenced the availability of MeHg to some extent, total DOM concentration was the most important factor in determining algal bioaccumulation of MeHg.

Microbial communities mediating algal detritus turnover under anaerobic conditions

PubMed Central

Morrison, Jessica M.; Murphy, Chelsea L.; Baker, Kristina; Zamor, Richard M.; Nikolai, Steve J.; Wilder, Shawn; Elshahed, Mostafa S.

2017-01-01

Background Algae encompass a wide array of photosynthetic organisms that are ubiquitously distributed in aquatic and terrestrial habitats. Algal species often bloom in aquatic ecosystems, providing a significant autochthonous carbon input to the deeper anoxic layers in stratified water bodies. In addition, various algal species have been touted as promising candidates for anaerobic biogas production from biomass. Surprisingly, in spite of its ecological and economic relevance, the microbial community involved in algal detritus turnover under anaerobic conditions remains largely unexplored. Results Here, we characterized the microbial communities mediating the degradation of Chlorella vulgaris (Chlorophyta), Chara sp. strain IWP1 (Charophyceae), and kelp Ascophyllum nodosum (phylum Phaeophyceae), using sediments from an anaerobic spring (Zodlteone spring, OK; ZDT), sludge from a secondary digester in a local wastewater treatment plant (Stillwater, OK; WWT), and deeper anoxic layers from a seasonally stratified lake (Grand Lake O’ the Cherokees, OK; GL) as inoculum sources. Within all enrichments, the majority of algal biomass was metabolized within 13–16 weeks, and the process was accompanied by an increase in cell numbers and a decrease in community diversity. Community surveys based on the V4 region of the 16S rRNA gene identified different lineages belonging to the phyla Bacteroidetes, Proteobacteria (alpha, delta, gamma, and epsilon classes), Spirochaetes, and Firmicutes that were selectively abundant under various substrate and inoculum conditions. Within all kelp enrichments, the microbial communities structures at the conclusion of the experiment were highly similar regardless of the enrichment source, and were dominated by the genus Clostridium, or family Veillonellaceae within the Firmicutes. In all other enrichments the final microbial community was dependent on the inoculum source, rather than the type of algae utilized as substrate. Lineages enriched

Didymosphenia geminata: Algal blooms in oligotrophic streams and rivers

USGS Publications Warehouse

Sundareshwar, P.V.; Upadhayay, S.; Abessa, M.; Honomichl, S.; Berdanier, B.; Spaulding, S.A.; Sandvik, C.; Trennepohl, A.

2011-01-01

In recent decades, the diatom Didymosphenia geminata has emerged as nuisance species in river systems around the world. This periphytic alga forms large "blooms" in temperate streams, presenting a counterintuitive result: the blooms occur primarily in oligotrophic streams and rivers, where phosphorus (P) availability typically limits primary production. The goal of this study is to examine how high algal biomass is formed under low P conditions. We reveal a biogeochemical process by which D. geminata mats concentrate P from flowing waters. First, the mucopolysaccaride stalks of D. geminata adsorb both iron (Fe) and P. Second, enzymatic and bacterial processes interact with Fe to increase the biological availability of P. We propose that a positive feedback between total stalk biomass and high growth rate is created, which results in abundant P for cell division. The affinity of stalks for Fe in association with iron-phosphorus biogeochemistry suggest a resolution to the paradox of algal blooms in oliogotrophic streams and rivers. Copyright 2011 by the American Geophysical Union.

Biodiesel from wastewater: lipid production in high rate algal pond receiving disinfected effluent.

PubMed

Assemany, Paula Peixoto; Calijuri, Maria Lucia; do Couto, Eduardo de Aguiar; Santiago, Aníbal Fonseca; Dos Reis, Alberto José Delgado

2015-01-01

The production of different species of microalgae in consortium with other micro-organisms from wastewaters may represent an alternative process, to reduce the costs, for obtaining biofuels. The aim of this study was to evaluate the influence of pre-ultraviolet disinfection (UV) in the production of lipids from biomass produced in high rate ponds. Two high rate algal ponds were evaluated: a pond that received domestic sewage without disinfection and the other receiving domestic sewage previously disinfected by UV radiation (uvHRAP). The UV disinfection did not lead to significant differences in fatty acid profile and total lipid productivities, although it increased algal biomass concentration and productivity as well as lipid content. Moreover, the overall biomass concentrations and productivities decreased with the UV disinfection, mostly as a consequence of a loss in bacterial load. We thus conclude that uvHRAP disinfection may represent a potential strategy to promote the cleaner and safer growth of algal biomass when cultivated in consortium with other micro-organisms. Mainly regarding the use of wastewater as culture medium, together with a cheaper production of lipids for biodiesel, pre-disinfection may represent an advance since extraction costs could be significantly trimmed due to the increase in lipid content.

Longitudinal analysis of microbiota in microalga Nannochloropsis salina cultures

DOE PAGES

Geng, Haifeng; Sale, Kenneth L.; Tran-Gyamfi, Mary Bao; ...

2016-03-08

Here, large-scale open microalgae cultivation has tremendous potential to make a significant contribution to replacing petroleum-based fuels with biofuels. Open algal cultures are unavoidably inhabited with a diversity of microbes that live on, influence, and shape the fate of these ecosystems. However, there is little understanding of the resilience and stability of the microbial communities in engineered semicontinuous algal systems. To evaluate the dynamics and resilience of the microbial communities in microalgae biofuel cultures, we conducted a longitudinal study on open systems to compare the temporal profiles of the microbiota from two multigenerational algal cohorts, which include one seeded withmore » the microbiota from an in-house culture and the other exogenously seeded with a natural-occurring consortia of bacterial species harvested from the Pacific Ocean. From these month-long, semicontinuous open microalga Nannochloropsis salina cultures, we sequenced a time-series of 46 samples, yielding 8804 operational taxonomic units derived from 9,160,076 high-quality partial 16S rRNA sequences. We provide quantitative evidence that clearly illustrates the development of microbial community is associated with microbiota ancestry. In addition, N. salina growth phases were linked with distinct changes in microbial phylotypes. Alteromonadeles dominated the community in the N. salina exponential phase whereas Alphaproteobacteria and Flavobacteriia were more prevalent in the stationary phase. We also demonstrate that the N. salina-associated microbial community in open cultures is diverse, resilient, and dynamic in response to environmental perturbations. This knowledge has general implications for developing and testing design principles of cultivated algal systems.« less

The Regulation of Gene Expression in Cnidarian-Algal Associations.

DTIC Science & Technology

1999-04-29

initiation, establishment and maintenance of cnidarian -algal-algal associations. These associations are of global significance as corals and other related...underlying the establishment of the cnidarian -algal partnership, Further, the work described the natural life history of two associations, chosen for...histories of two cnidarians (hosts), a tropical coral Fungia scutaria and a temperate anemone Anthopleura elegantissima. We examined symbiosis onset in

[Primary culture of human normal epithelial cells].

PubMed

Tang, Yu; Xu, Wenji; Guo, Wanbei; Xie, Ming; Fang, Huilong; Chen, Chen; Zhou, Jun

2017-11-28

The traditional primary culture methods of human normal epithelial cells have disadvantages of low activity of cultured cells, the low cultivated rate and complicated operation. To solve these problems, researchers made many studies on culture process of human normal primary epithelial cell. In this paper, we mainly introduce some methods used in separation and purification of human normal epithelial cells, such as tissue separation method, enzyme digestion separation method, mechanical brushing method, red blood cell lysis method, percoll layered medium density gradient separation method. We also review some methods used in the culture and subculture, including serum-free medium combined with low mass fraction serum culture method, mouse tail collagen coating method, and glass culture bottle combined with plastic culture dish culture method. The biological characteristics of human normal epithelial cells, the methods of immunocytochemical staining, trypan blue exclusion are described. Moreover, the factors affecting the aseptic operation, the conditions of the extracellular environment, the conditions of the extracellular environment during culture, the number of differential adhesion, and the selection and dosage of additives are summarized.

Algal Biofuels Techno-Economic Analysis | Bioenergy | NREL

Science.gov Websites

Biofuels Techno-Economic Analysis Algal Biofuels Techno-Economic Analysis To promote an understanding of the challenges and opportunities unique to microalgae, NREL's Algae Techno-Economic Analysis group focuses on techno-economic analysis (TEA) for the production and conversion of algal biomass into

Reversible gelling culture media for in-vitro cell culture in three-dimensional matrices

DOEpatents

An, Yuehuei H.; Mironov, Vladimir A.; Gutowska, Anna

2000-01-01

A gelling cell culture medium useful for forming a three dimensional matrix for cell culture in vitro is prepared by copolymerizing an acrylamide derivative with a hydrophilic comonomer to form a reversible (preferably thermally reversible) gelling linear random copolymer in the form of a plurality of linear chains having a plurality of molecular weights greater than or equal to a minimum gelling molecular weight cutoff, mixing the copolymer with an aqueous solvent to form a reversible gelling solution and adding a cell culture medium to the gelling solution to form the gelling cell culture medium. Cells such as chondrocytes or hepatocytes are added to the culture medium to form a seeded culture medium, and temperature of the medium is raised to gel the seeded culture medium and form a three dimensional matrix containing the cells. After propagating the cells in the matrix, the cells may be recovered by lowering the temperature to dissolve the matrix and centrifuging.

Algicidal Effects of a Novel Marine Pseudoalteromonas Isolate (Class Proteobacteria, Gamma Subdivision) on Harmful Algal Bloom Species of the Genera Chattonella, Gymnodinium, and Heterosigma

PubMed Central

Lovejoy, Connie; Bowman, John P.; Hallegraeff, Gustaaf M.

1998-01-01

During a bacterial survey of the Huon Estuary in southern Tasmania, Australia, we isolated a yellow-pigmented Pseudoalteromonas strain (class Proteobacteria, gamma subdivision), designated strain Y, that had potent algicidal effects on harmful algal bloom species. This organism was identified by 16S rRNA sequencing as a strain with close affinities to Pseudoalteromonas peptidysin. This bacterium caused rapid cell lysis and death (within 3 h) of gymnodinoids (including Gymnodinium catenatum) and raphidophytes (Chattonella marina and Heterosigma akashiwo). It caused ecdysis of armored dinoflagellates (e.g., Alexandrium catenella, Alexandrium minutum, and Prorocentrum mexicanum), but the algal cultures then recovered over the subsequent 24 h. Strain Y had no effect on a cryptomonad (Chroomonas sp.), a diatom (Skeletonema sp.), a cyanobacterium (Oscillatoria sp.), and two aplastidic protozoans. The algicidal principle of strain Y was excreted into the seawater medium and lost its efficacy after heating. Another common bacterial species, Pseudoalteromonas carrageenovora, was isolated at the same time and did not have these algicidal effects. The minimum concentrations of strain Y required to kill G. catenatum were higher than the mean concentrations found in nature under nonbloom conditions. However, the new bacterium showed a chemotactic, swarming behavior that resulted in localized high concentrations around target organisms. These observations imply that certain bacteria could play an important role in regulating the onset and development of harmful algal blooms. PMID:9687434

Traditional and Modern Cell Culture in Virus Diagnosis.

PubMed

Hematian, Ali; Sadeghifard, Nourkhoda; Mohebi, Reza; Taherikalani, Morovat; Nasrolahi, Abbas; Amraei, Mansour; Ghafourian, Sobhan

2016-04-01

Cell cultures are developed from tissue samples and then disaggregated by mechanical, chemical, and enzymatic methods to extract cells suitable for isolation of viruses. With the recent advances in technology, cell culture is considered a gold standard for virus isolation. This paper reviews the evolution of cell culture methods and demonstrates why cell culture is a preferred method for identification of viruses. In addition, the advantages and disadvantages of both traditional and modern cell culture methods for diagnosis of each type of virus are discussed. Detection of viruses by the novel cell culture methods is considered more accurate and sensitive. However, there is a need to include some more accurate methods such as molecular methods in cell culture for precise identification of viruses.

Algal sludge from Taihu Lake can be utilized to create novel PGPR-containing bio-organic fertilizers.

PubMed

Zhang, Miao; Li, Rong; Cao, Liangliang; Shi, Juanjuan; Liu, Hongjun; Huang, Yan; Shen, Qirong

2014-01-01

Large amounts of refloated algal sludge from Taihu Lake result in secondary environmental pollution due to annual refloatation. This study investigated the possibility to produce bio-organic fertilizer (BIO) using algal sludge as a solid-state fermentation (SSF) medium. Results showed that addition of algal sludge contributed to efficient SFF by a plant growth-promoting rhizobacteria (PGPR) strain SQR9 and improved the nutrient contents in the novel BIO. The optimum water content and initial inoculation size were 45% and 5%, respectively. After 6 days of SSF, the biomass of strain SQR9 was increased to a cell density of more than 5 × 10(7) CFU g(-1). Microcystins were rapidly degraded, and a high germination index value was observed. Plant growth experiments showed that the produced BIO efficiently promoted plant growth. Additional testing showed that the novel SSF process was also suitable for other PGPR strains. This study provides a novel way of high-value utilization of algal sludge from Taihu Lake by producing low-cost but high-quality BIOs. Copyright © 2013 Elsevier Ltd. All rights reserved.

Algicidal bacteria in the sea and their impact on algal blooms.

PubMed

Mayali, Xavier; Azam, Farooq

2004-01-01

Over the past two decades, many reports have revealed the existence of bacteria capable of killing phytoplankton. These algicidal bacteria sometimes increase in abundance concurrently with the decline of algal blooms, suggesting that they may affect algal bloom dynamics. Here, we synthesize the existing knowledge on algicidal bacteria interactions with marine eukaryotic microalgae. We discuss the effectiveness of the current methods to characterize the algicidal phenotype in an ecosystem context. We briefly consider the literature on the phylogenetic identification of algicidal bacteria, their interaction with their algal prey, the characterization of algicidal molecules, and the enumeration of algicidal bacteria during algal blooms. We conclude that, due to limitations of current methods, the evidence for algicidal bacteria causing algal bloom decline is circumstantial. New methods and an ecosystem approach are needed to test hypotheses on the impact of algicidal bacteria in algal bloom dynamics. This will require enlarging the scope of inquiry from its current focus on the potential utility of algicidal bacteria in the control of harmful algal blooms. We suggest conceptualizing bacterial algicidy within the general problem of bacterial regulation of algal community structure in the ocean.

Application of cell co-culture system to study fat and muscle cells.

PubMed

Pandurangan, Muthuraman; Hwang, Inho

2014-09-01

Animal cell culture is a highly complex process, in which cells are grown under specific conditions. The growth and development of these cells is a highly unnatural process in vitro condition. Cells are removed from animal tissues and artificially cultured in various culture vessels. Vitamins, minerals, and serum growth factors are supplied to maintain cell viability. Obtaining result homogeneity of in vitro and in vivo experiments is rare, because their structure and function are different. Living tissues have highly ordered complex architecture and are three-dimensional (3D) in structure. The interaction between adjacent cell types is quite distinct from the in vitro cell culture, which is usually two-dimensional (2D). Co-culture systems are studied to analyze the interactions between the two different cell types. The muscle and fat co-culture system is useful in addressing several questions related to muscle modeling, muscle degeneration, apoptosis, and muscle regeneration. Co-culture of C2C12 and 3T3-L1 cells could be a useful diagnostic tool to understand the muscle and fat formation in animals. Even though, co-culture systems have certain limitations, they provide a more realistic 3D view and information than the individual cell culture system. It is suggested that co-culture systems are useful in evaluating the intercellular communication and composition of two different cell types.

Neonatal rat heart cells cultured in simulated microgravity

NASA Technical Reports Server (NTRS)

Akins, Robert E.; Schroedl, Nancy A.; Gonda, Steve R.; Hartzell, Charles R.

1994-01-01

In vitro characteristics of cardiac cells cultured in simulated microgravity are reported. Tissue culture methods performed at unit gravity constrain cells to propagate, differentiate, and interact in a two dimensional (2D) plane. Neonatal rat cardiac cells in 2D culture organize predominantly as bundles of cardiomyocytes with the intervening areas filled by non-myocyte cell types. Such cardiac cell cultures respond predictably to the addition of exogenous compounds, and in many ways they represent an excellent in vitro model system. The gravity-induced 2D organization of the cells, however, does not accurately reflect the distribution of cells in the intact tissue. We have begun characterizations of a three-dimensional (3D) culturing system designed to mimic microgravity. The NASA designed High-Aspect-Ratio-Vessel (HARV) bioreactors provide a low shear environment which allows cells to be cultured in static suspension. HARV-3D cultures were prepared on microcarrier beads and compared to control-2D cultures using a combination of microscopic and biochemical techniques. Both systems were uniformly inoculated and medium exchanged at standard intervals. Cells in control cultures adhered to the polystyrene surface of the tissue culture dishes and exhibited typical 2D organization. Cells in cultured in HARV's adhered to microcarrier beads, the beads aggregated into defined clusters containing 8 to 15 beads per cluster, and the clusters exhibited distinct 3D layers: myocytes and fibroblasts appeared attached to the surfaces of beads and were overlaid by an outer cell type. In addition, cultures prepared in HARV's using alternative support matrices also displayed morphological formations not seen in control cultures. Generally, the cells prepared in HARV and control cultures were similar, however, the dramatic alterations in 3D organization recommend the HARV as an ideal vessel for the generation of tissue-like organizations of cardiac cells in simulated microgravity.

Neonatal rat heart cells cultured in simulated microgravity

NASA Technical Reports Server (NTRS)

Akins, R. E.; Schroedl, N. A.; Gonda, S. R.; Hartzell, C. R.

1997-01-01

In vitro characteristics of cardiac cells cultured in simulated microgravity are reported. Tissue culture methods performed at unit gravity constrain cells to propagate, differentiate, and interact in a two-dimensional (2D) plane. Neonatal rat cardiac cells in 2D culture organize predominantly as bundles of cardiomyocytes with the intervening areas filled by nonmyocyte cell types. Such cardiac cell cultures respond predictably to the addition of exogenous compounds, and in many ways they represent an excellent in vitro model system. The gravity-induced 2D organization of the cells, however, does not accurately reflect the distribution of cells in the intact tissue. We have begun characterizations of a three-dimensional (3D) culturing system designed to mimic microgravity. The NASA-designed High-Aspect Ratio Vessel (HARV) bioreactors provide a low shear environment that allows cells to be cultured in static suspension. HARV-3D cultures were prepared on microcarrier beads and compared to control-2D cultures using a combination of microscopic and biochemical techniques. Both systems were uniformly inoculated and medium exchanged at standard intervals. Cells in control cultures adhered to the polystyrene surface of the tissue culture dishes and exhibited typical 2D organization. Cells cultured in HARVs adhered to microcarrier beads, the beads aggregated into defined clusters containing 8 to 15 beads per cluster, and the clusters exhibited distinct 3D layers: myocytes and fibroblasts appeared attached to the surfaces of beads and were overlaid by an outer cell type. In addition, cultures prepared in HARVs using alternative support matrices also displayed morphological formations not seen in control cultures. Generally, the cells prepared in HARV and control cultures were similar; however, the dramatic alterations in 3D organization recommend the HARV as an ideal vessel for the generation of tissuelike organization of cardiac cells in vitro.

Differentiation of mammalian skeletal muscle cells cultured on microcarrier beads in a rotating cell culture system

NASA Technical Reports Server (NTRS)

Torgan, C. E.; Burge, S. S.; Collinsworth, A. M.; Truskey, G. A.; Kraus, W. E.

2000-01-01

The growth and repair of adult skeletal muscle are due in part to activation of muscle precursor cells, commonly known as satellite cells or myoblasts. These cells are responsive to a variety of environmental cues, including mechanical stimuli. The overall goal of the research is to examine the role of mechanical signalling mechanisms in muscle growth and plasticity through utilisation of cell culture systems where other potential signalling pathways (i.e. chemical and electrical stimuli) are controlled. To explore the effects of decreased mechanical loading on muscle differentiation, mammalian myoblasts are cultured in a bioreactor (rotating cell culture system), a model that has been utilised to simulate microgravity. C2C12 murine myoblasts are cultured on microcarrier beads in a bioreactor and followed throughout differentiation as they form a network of multinucleated myotubes. In comparison with three-dimensional control cultures that consist of myoblasts cultured on microcarrier beads in teflon bags, myoblasts cultured in the bioreactor exhibit an attenuation in differentiation. This is demonstrated by reduced immunohistochemical staining for myogenin and alpha-actinin. Western analysis shows a decrease, in bioreactor cultures compared with control cultures, in levels of the contractile proteins myosin (47% decrease, p < 0.01) and tropomyosin (63% decrease, p < 0.01). Hydrodynamic measurements indicate that the decrease in differentiation may be due, at least in part, to fluid stresses acting on the myotubes. In addition, constraints on aggregate size imposed by the action of fluid forces in the bioreactor affect differentiation. These results may have implications for muscle growth and repair during spaceflight.

Culturing primary mouse pancreatic ductal cells.

PubMed

Reichert, Maximilian; Rhim, Andrew D; Rustgi, Anil K

2015-06-01

The most common subtype of pancreatic cancer is pancreatic ductal adenocarcinoma (PDAC). PDAC resembles ductal cells morphologically. To study pancreatic ductal cell (PDC) and pancreatic intraepithelial neoplasia (PanIN)/PDAC biology, it is essential to have reliable in vitro culture conditions. Here we describe a methodology to isolate, culture, and passage PDCs and duct-like cells from the mouse pancreas. It can be used to isolate cells from genetically engineered mouse models (GEMMs), providing a valuable tool to study disease models in vitro to complement in vivo findings. The culture conditions allow epithelial cells to outgrow fibroblast and other "contaminating" cell types within a few passages. However, the resulting cultures, although mostly epithelial, are not completely devoid of fibroblasts. Regardless, this protocol provides guidelines for a robust in vitro culture system to isolate, maintain, and expand primary pancreatic ductal epithelial cells. It can be applied to virtually all GEMMs of pancreatic disease and other diseases and cancers that arise from ductal structures. Because most carcinomas resemble ductal structures, this protocol has utility in the study of other cancers in addition to PDAC, such as breast and prostate cancers. © 2015 Cold Spring Harbor Laboratory Press.

Oscillating Cell Culture Bioreactor

NASA Technical Reports Server (NTRS)

Freed, Lisa E.; Cheng, Mingyu; Moretti, Matteo G.

2010-01-01

To better exploit the principles of gas transport and mass transport during the processes of cell seeding of 3D scaffolds and in vitro culture of 3D tissue engineered constructs, the oscillatory cell culture bioreactor provides a flow of cell suspensions and culture media directly through a porous 3D scaffold (during cell seeding) and a 3D construct (during subsequent cultivation) within a highly gas-permeable closed-loop tube. This design is simple, modular, and flexible, and its component parts are easy to assemble and operate, and are inexpensive. Chamber volume can be very low, but can be easily scaled up. This innovation is well suited to work with different biological specimens, particularly with cells having high oxygen requirements and/or shear sensitivity, and different scaffold structures and dimensions. The closed-loop changer is highly gas permeable to allow efficient gas exchange during the cell seeding/culturing process. A porous scaffold, which may be seeded with cells, is fixed by means of a scaffold holder to the chamber wall with scaffold/construct orientation with respect to the chamber determined by the geometry of the scaffold holder. A fluid, with/without biological specimens, is added to the chamber such that all, or most, of the air is displaced (i.e., with or without an enclosed air bubble). Motion is applied to the chamber within a controlled environment (e.g., oscillatory motion within a humidified 37 C incubator). Movement of the chamber induces relative motion of the scaffold/construct with respect to the fluid. In case the fluid is a cell suspension, cells will come into contact with the scaffold and eventually adhere to it. Alternatively, cells can be seeded on scaffolds by gel entrapment prior to bioreactor cultivation. Subsequently, the oscillatory cell culture bioreactor will provide efficient gas exchange (i.e., of oxygen and carbon dioxide, as required for viability of metabolically active cells) and controlled levels of fluid

Variations of algal communities cause darkening of a Greenland glacier.

PubMed

Lutz, Stefanie; Anesio, Alexandre M; Jorge Villar, Susana E; Benning, Liane G

2014-08-01

We have assessed the microbial ecology on the surface of Mittivakkat glacier in SE-Greenland during the exceptional high melting season in July 2012 when the so far most extreme melting rate for the Greenland Ice Sheet has been recorded. By employing a complementary and multi-disciplinary field sampling and analytical approach, we quantified the dramatic changes in the different microbial surface habitats (green snow, red snow, biofilms, grey ice, cryoconite holes). The observed clear change in dominant algal community and their rapidly changing cryo-organic adaptation inventory was linked to the high melting rate. The changes in carbon and nutrient fluxes between different microbial pools (from snow to ice, cryoconite holes and glacial forefronts) revealed that snow and ice algae dominate the net primary production at the onset of melting, and that they have the potential to support the cryoconite hole communities as carbon and nutrient sources. A large proportion of algal cells is retained on the glacial surface and temporal and spatial changes in pigmentation contribute to the darkening of the snow and ice surfaces. This implies that the fast, melt-induced algal growth has a high albedo reduction potential, and this may lead to a positive feedback speeding up melting processes. © 2014 Federation of European Microbiological Societies. Published by John Wiley & Sons Ltd. All rights reserved.

Regulation of gametogenesis and zoosporogenesis in Ulva linza (Chlorophyta): comparison with Ulva mutabilis and potential for laboratory culture.

PubMed

Vesty, Eleanor F; Kessler, Ralf W; Wichard, Thomas; Coates, Juliet C

2015-01-01

Green Ulvophyte macroalgae represent attractive model systems for understanding growth, development, and evolution. They are untapped resources for food, fuel, and high-value compounds, but can also form nuisance blooms. To fully analyze green seaweed morphogenesis, controlled laboratory-based culture of these organisms is required. To date, only a single Ulvophyte species, Ulva mutabilis Føyn, has been manipulated to complete its whole life cycle in laboratory culture and to grow continuously under axenic conditions. Such cultures are essential to address multiple key questions in Ulva development and in algal-bacterial interactions. Here we show that another Ulva species, U. linza, with a broad geographical distribution, has the potential to be grown in axenic culture similarly to U. mutabilis. U. linza can be reliably induced to sporulate (form gametes and zoospores) in the laboratory, by cutting the relevant thallus tissue into small pieces and removing extracellular inhibitors (sporulation and swarming inhibitors). The germ cells work as an ideal feed stock for standardized algae cultures. The requirement of U. linza for bacterial signals to induce its normal morphology (particularly of the rhizoids) appears to have a species-specific component. The axenic cultures of these two species pave the way for future comparative studies of algal-microbial interactions.

A continuous perfusion microplate for cell culture.

PubMed

Goral, Vasiliy N; Zhou, Chunfeng; Lai, Fang; Yuen, Po Ki

2013-03-21

We describe a 96-well microplate with fluidically connected wells that enables the continuous fluid perfusion between wells without the need for external pumping. A single unit in such a perfusion microplate consists of three wells: a source well, a sample (cell culture) well in the middle and a waste well. Fluid perfusion is achieved using a combination of the hydrostatic pressure generated by different liquid levels in the wells and the fluid wicking through narrow strips of a cellulose membrane connecting the wells. There is an excellent correspondence between the observed perfusion flow dynamics and the flow simulations based on Darcy's Law. Hepatocytes (C3A cells) cultured for 4 days in the perfusion microplate with no media exchange in the cell culture well had the same viability as hepatocytes exposed to a daily exchange of media. EOC 20 cells that require media conditioned by LADMAC cells were shown to be equally viable in the adjacent cell culture well of the perfusion microplate with LADMAC cells cultured in the source well. Tegafur, a prodrug, when added to primary human hepatocytes in the source well, was metabolized into a cytotoxic metabolite that kills colon cancer cells (HCT 116) cultured in the adjacent cell culture well; no toxicity was observed when only medium was in the source well. These results suggest that the perfusion microplate is a useful tool for a variety of cell culture applications with benefits ranging from labor savings to enabling in vivo-like toxicity studies.

Pan-Arctic sea ice-algal chl a biomass and suitable habitat are largely underestimated for multiyear ice.

PubMed

Lange, Benjamin A; Flores, Hauke; Michel, Christine; Beckers, Justin F; Bublitz, Anne; Casey, John Alec; Castellani, Giulia; Hatam, Ido; Reppchen, Anke; Rudolph, Svenja A; Haas, Christian

2017-11-01

There is mounting evidence that multiyear ice (MYI) is a unique component of the Arctic Ocean and may play a more important ecological role than previously assumed. This study improves our understanding of the potential of MYI as a suitable habitat for sea ice algae on a pan-Arctic scale. We sampled sea ice cores from MYI and first-year sea ice (FYI) within the Lincoln Sea during four consecutive spring seasons. This included four MYI hummocks with a mean chl a biomass of 2.0 mg/m 2 , a value significantly higher than FYI and MYI refrozen ponds. Our results support the hypothesis that MYI hummocks can host substantial ice-algal biomass and represent a reliable ice-algal habitat due to the (quasi-) permanent low-snow surface of these features. We identified an ice-algal habitat threshold value for calculated light transmittance of 0.014%. Ice classes and coverage of suitable ice-algal habitat were determined from snow and ice surveys. These ice classes and associated coverage of suitable habitat were applied to pan-Arctic CryoSat-2 snow and ice thickness data products. This habitat classification accounted for the variability of the snow and ice properties and showed an areal coverage of suitable ice-algal habitat within the MYI-covered region of 0.54 million km 2 (8.5% of total ice area). This is 27 times greater than the areal coverage of 0.02 million km 2 (0.3% of total ice area) determined using the conventional block-model classification, which assigns single-parameter values to each grid cell and does not account for subgrid cell variability. This emphasizes the importance of accounting for variable snow and ice conditions in all sea ice studies. Furthermore, our results indicate the loss of MYI will also mean the loss of reliable ice-algal habitat during spring when food is sparse and many organisms depend on ice-algae. © 2017 The Authors. Global Change Biology Published by John Wiley & Sons Ltd.

Characterization and performance of anodic mixed culture biofilms in submersed microbial fuel cells.

PubMed

Saba, Beenish; Christy, Ann D; Yu, Zhongtang; Co, Anne C; Islam, Rafiq; Tuovinen, Olli H

2017-02-01

Microbial fuel cells (MFCs) were designed for laboratory scale experiments to study electroactive biofilms in anodic chambers. Anodic biofilms and current generation during biofilm growth were examined using single chambered MFCs submersed in algal catholyte. A culture of the marine green alga Nanochloropsis salina was used as a biocatholyte, and a rumen fluid microbiota was the anodic chamber inoculum. Electrical impedance spectroscopy was performed under varying external resistance once a week to identify mass transport limitations at the biofilm-electrolyte interface during the four-week experiment. The power generation increased from 249 to 461mWm -2 during the time course. Confocal laser scanning microscopy imaging showed that the depth of the bacterial biofilm on the anode was about 65μm. There were more viable bacteria on the biofilm surface and near the biofilm-electrolyte interface as compared to those close to the anode surface. The results suggest that biofilm growth on the anode creates a conductive layer, which can help overcome mass transport limitations in MFCs. Copyright © 2016 Elsevier B.V. All rights reserved.

Basic techniques in mammalian cell tissue culture.

PubMed

Phelan, Katy; May, Kristin M

2015-03-02

Cultured mammalian cells are used extensively in cell biology studies. It requires a number of special skills in order to be able to preserve the structure, function, behavior, and biology of the cells in culture. This unit describes the basic skills required to maintain and preserve cell cultures: maintaining aseptic technique, preparing media with the appropriate characteristics, passaging, freezing and storage, recovering frozen stocks, and counting viable cells. Copyright © 2015 John Wiley & Sons, Inc.

Polydimethylsiloxane SlipChip for mammalian cell culture applications.

PubMed

Chang, Chia-Wen; Peng, Chien-Chung; Liao, Wei-Hao; Tung, Yi-Chung

2015-11-07

This paper reports a polydimethylsiloxane (PDMS) SlipChip for in vitro cell culture applications, multiple-treatment assays, cell co-cultures, and cytokine detection assays. The PDMS SlipChip is composed of two PDMS layers with microfluidic channels on each surface that are separated by a thin silicone fluid (Si-fluid) layer. The integration of Si-fluid enables the two PDMS layers to be slid to different positions; therefore, the channel patterns can be re-arranged for various applications. The SlipChip design significantly reduces the complexity of sample handling, transportation, and treatment processes. To apply the developed SlipChip for cell culture applications, human lung adenocarcinoma epithelial cells (A549) and lung fibroblasts (MRC-5) were cultured to examine the biocompatibility of the developed PDMS SlipChip. Moreover, embryonic pluripotent stem cells (ES-D3) were also cultured in the device to evaluate the retention of their stemness in the device. The experimental results show that cell morphology, viability and proliferation are not affected when the cells are cultured in the SlipChip, indicating that the device is highly compatible with mammalian cell culture. In addition, the stemness of the ES-D3 cells was highly retained after they were cultured in the device, suggesting the feasibility of using the SlipChip for stem cell research. Various cell experiments, such as simultaneous triple staining of cells and co-culture of MRC-5 with A549 cells, were also performed to demonstrate the functionalities of the PDMS SlipChip. Furthermore, we used a cytokine detection assay to evaluate the effect of endotoxin (lipopolysaccharides, LPS) treatment on the cytokine secretion of A549 cells using the SlipChip. The developed PDMS SlipChip provides a straightforward and effective platform for various on-chip in vitro cell cultures and consequent analysis, which is promising for a number of cell biology studies and biomedical applications.

Implementation of UV-based advanced oxidation processes in algal medium recycling.

PubMed

Wang, Wenxuan; Sha, Jun; Lu, Zhiying; Shao, Senlin; Sun, Peizhe; Hu, Qiang; Zhang, Xuezhi

2018-09-01

Algae show great potential as sustainable feedstock for numerous bioproducts. However, large volume of water consumption during algal biomass production makes that the culture media recycling is a necessity due to economic and environmental concern. To avoid the negative effect of enriched organic matters in the harvested culture media, pre-treatment prior to medium replenishment and reuse is required. In this study, degradation of algenitic organic matters (AOM) in the culture media by UV-based photolysis processes (i.e., direct UV, UV/peroxydisulfate (PDS), UV/H 2 O 2 , and UV/NH 2 Cl) was explored. The results showed that UV, UV/PDS, UV/H 2 O 2 and UV/NH 2 Cl caused a decrease of SUVA for 29.9%, 35.4%, 40.45%, and 22.6%, respectively, though the organic matter was almost not mineralized. Fluorescence excitation-emission matrix combined with parallel factor analysis indicated that UV/PDS and UV/H 2 O 2 degraded 47.26%-56.31% of the fulvic-like and humic-like fractions in AOM. Powder activated carbon absorption and growth evaluation for the AOPs-treated media indicated that UV/PDS and UV/H 2 O 2 processes not only could remove the growth inhibitors in the media, but were also beneficial to the algae growth. These results suggested that UV/PDS and UV/H 2 O 2 could effectively degrade the hydrophobic components in AOM and converted the growth inhibition fraction of AOM in the recycled media into nutrient source for algal growth. Different from the general application of UV-based AOP in the wastewater treatment, this study provided an innovative idea about how to pre-treat AOM in the media recycling: utilization rather than removal, which was a more sustainable and environment-friendly technology. Copyright © 2018 Elsevier B.V. All rights reserved.

Long-term culture and differentiation of porcine red bone marrow hematopoietic cells co-cultured with immortalized mesenchymal cells.

PubMed

Garba, Abubakar; Acar, Delphine D; Roukaerts, Inge D M; Desmarets, Lowiese M B; Devriendt, Bert; Nauwynck, Hans J

2017-09-01

Mesenchymal cells are multipotent stromal cells with self-renewal, differentiation and immunomodulatory capabilities. We aimed to develop a co-culture model for differentiating hematopoietic cells on top of immortalized mesenchymal cells for studying interactions between hematopoietic and mesenchymal cells, useful for adequately exploring the therapeutic potential of mesenchymal cells. In this study, we investigated the survival, proliferation and differentiation of porcine red bone marrow hematopoietic cells co-cultured with immortalized porcine bone marrow mesenchymal cells for a period of five weeks. Directly after collection, primary porcine bone marrow mesenchymal cells adhered firmly to the bottom of the culture plates and showed a fibroblast-like appearance, one week after isolation. Upon immortalization, porcine bone marrow mesenchymal cells were continuously proliferating. They were positive for simian virus 40 (SV40) large T antigen and the mesenchymal cell markers CD44 and CD55. Isolated red bone marrow cells were added to these immortalized mesenchymal cells. Five weeks post-seeding, 92±6% of the red bone marrow hematopoietic cells were still alive and their number increased 3-fold during five weekly subpassages on top of the immortalized mesenchymal cells. The red bone marrow hematopoietic cells were originally small and round; later, the cells increased in size. Some of them became elongated, while others remained round. Tiny dendrites appeared attaching hematopoietic cells to the underlying immortalized mesenchymal cells. Furthermore, weekly differential-quick staining of the cells indicated the presence of monoblasts, monocytes, macrophages and lymphocytes in the co-cultures. At three weeks of co-culture, flow cytometry analysis showed an increased surface expression of CD172a, CD14, CD163, CD169, CD4 and CD8 up to 37±0.8%, 40±8%, 41±4%, 23±3% and 19±5% of the hematopoietic cells, respectively. In conclusion, continuous mesenchymal cell

Comprehensive computational model for combining fluid hydrodynamics, light transport and biomass growth in a Taylor vortex algal photobioreactor: Lagrangian approach.

PubMed

Gao, Xi; Kong, Bo; Vigil, R Dennis

2017-01-01

A comprehensive quantitative model incorporating the effects of fluid flow patterns, light distribution, and algal growth kinetics on biomass growth rate is developed in order to predict the performance of a Taylor vortex algal photobioreactor for culturing Chlorella vulgaris. A commonly used Lagrangian strategy for coupling the various factors influencing algal growth was employed whereby results from computational fluid dynamics and radiation transport simulations were used to compute numerous microorganism light exposure histories, and this information in turn was used to estimate the global biomass specific growth rate. The simulations provide good quantitative agreement with experimental data and correctly predict the trend in reactor performance as a key reactor operating parameter is varied (inner cylinder rotation speed). However, biomass growth curves are consistently over-predicted and potential causes for these over-predictions and drawbacks of the Lagrangian approach are addressed. Copyright © 2016 Elsevier Ltd. All rights reserved.

3D Cell Culture in Alginate Hydrogels

PubMed Central

Andersen, Therese; Auk-Emblem, Pia; Dornish, Michael

2015-01-01

This review compiles information regarding the use of alginate, and in particular alginate hydrogels, in culturing cells in 3D. Knowledge of alginate chemical structure and functionality are shown to be important parameters in design of alginate-based matrices for cell culture. Gel elasticity as well as hydrogel stability can be impacted by the type of alginate used, its concentration, the choice of gelation technique (ionic or covalent), and divalent cation chosen as the gel inducing ion. The use of peptide-coupled alginate can control cell–matrix interactions. Gelation of alginate with concomitant immobilization of cells can take various forms. Droplets or beads have been utilized since the 1980s for immobilizing cells. Newer matrices such as macroporous scaffolds are now entering the 3D cell culture product market. Finally, delayed gelling, injectable, alginate systems show utility in the translation of in vitro cell culture to in vivo tissue engineering applications. Alginate has a history and a future in 3D cell culture. Historically, cells were encapsulated in alginate droplets cross-linked with calcium for the development of artificial organs. Now, several commercial products based on alginate are being used as 3D cell culture systems that also demonstrate the possibility of replacing or regenerating tissue. PMID:27600217

Testicular Sertoli cells influence the proliferation and immunogenicity of co-cultured endothelial cells

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

Fan, Ping, E-mail: fanpinggoodluck@163.com; He, Lan; Pu, Dan

Research highlights: {yields} The proliferation of dramatic increased by co-cultured with Sertoli cells. {yields} VEGF receptor-2 expression of ECs was up-regulated by co-cultured with Sertoli cells. {yields} The MHC expression of ECs induced by INF-{gamma} and IL-6, IL-8 and sICAM induced by TNF-{alpha} decreased respectively after co-cultured with Sertoli cells. {yields} ECs co-cultured with Sertoli cells also didn't increase the stimulation index of spleen lymphocytes. -- Abstract: The major problem of the application of endothelial cells (ECs) in transplantation is the lack of proliferation and their immunogenicity. In this study, we co-cultured ECs with Sertoli cells to monitor whether Sertolimore » cells can influence the proliferation and immunogenicity of co-cultured ECs. Sertoli cells were isolated from adult testicular tissue. ECs were divided into the control group and the experimental group, which included three sub-groups co-cultured with 1 x 10{sup 3}, 1 x 10{sup 4} or 1 x 10{sup 5} cell/ml of Sertoli cells. The growth and proliferation of ECs were observed microscopically, and the expression of vascular endothelial growth factor (VEGF) receptor-2 (KDR) was examined by Western blotting. In another experiment, ECs were divided into the control group, the single culture group and the co-culture group with the optimal concentration of Sertoli cells. After INF-{gamma} and TNF-{alpha} were added to the culture medium, MHC II antigen expression was detected by immunofluorescence staining and western blotting; interleukin (IL)-6, IL-8 and soluble intercellular adhesion molecule (sICAM) were measured in the culture medium by ELISA. We demonstrated that 1 x 10{sup 4} cell/ml Sertoli cells promoted the proliferation of co-cultured ECs more dramatically than that in other groups (P < 0.05). Western blotting showed that 1 x 10{sup 4} cell/ml of the Sertoli cells was most effective in the up-regulation of KDR expression in the co-cultured ECs (P < 0.05). Sertoli

Screening of surfactants for harmful algal blooms mitigation.

PubMed

Sun, Xiao-Xia; Han, Kyung-Nam; Choi, Joong-Ki; Kim, Eun-Ki

2004-05-01

Screening experiments were conducted in order to find promising synthetic surfactants for harmful algal blooms (HABs) mitigation. The chemically synthesized surfactant cocamidopropyl betaine (CAPB) showed characteristics of relatively high inhibition efficiency, high biodegradability and low cost. The motility inhibition ratios of 10 mg/L CAPB on Cochlodinium polykrikoides and Alexandrium tamarense were about 60% after 5 min. The biodegradation test indicated that the half-life of CAPB in seawater was shorter than one day and 90% was biodegraded after five days under the initial concentration of 100 mg/L at 25 degrees C. Further cell lysis experiments revealed the selective lysis effect of CAPB on different HAB organisms. More than 90% of C. polykrikoides lysed at the concentration of 10 mg/L CAPB after 24 h and at 15 mg/L CAPB after 4 h, whereas the lysis effect of CAPB on A. tamarense was slight, no more than 10% after 2 h interaction with 50 mg/L CAPB. This research provided preliminary data for CAPB as a candidate in harmful algal blooms mitigation and pointed out unresolved problems for its practical application in the meantime.

A Versatile Bioreactor for Dynamic Suspension Cell Culture. Application to the Culture of Cancer Cell Spheroids

PubMed Central

Madeddu, Denise; Cerino, Giulia; Falco, Angela; Frati, Caterina; Gallo, Diego; Deriu, Marco A.; Falvo D’Urso Labate, Giuseppe; Quaini, Federico; Audenino, Alberto; Morbiducci, Umberto

2016-01-01

A versatile bioreactor suitable for dynamic suspension cell culture under tunable shear stress conditions has been developed and preliminarily tested culturing cancer cell spheroids. By adopting simple technological solutions and avoiding rotating components, the bioreactor exploits the laminar hydrodynamics establishing within the culture chamber enabling dynamic cell suspension in an environment favourable to mass transport, under a wide range of tunable shear stress conditions. The design phase of the device has been supported by multiphysics modelling and has provided a comprehensive analysis of the operating principles of the bioreactor. Moreover, an explanatory example is herein presented with multiphysics simulations used to set the proper bioreactor operating conditions for preliminary in vitro biological tests on a human lung carcinoma cell line. The biological results demonstrate that the ultralow shear dynamic suspension provided by the device is beneficial for culturing cancer cell spheroids. In comparison to the static suspension control, dynamic cell suspension preserves morphological features, promotes intercellular connection, increases spheroid size (2.4-fold increase) and number of cycling cells (1.58-fold increase), and reduces double strand DNA damage (1.5-fold reduction). It is envisioned that the versatility of this bioreactor could allow investigation and expansion of different cell types in the future. PMID:27144306

A Versatile Bioreactor for Dynamic Suspension Cell Culture. Application to the Culture of Cancer Cell Spheroids.

PubMed

Massai, Diana; Isu, Giuseppe; Madeddu, Denise; Cerino, Giulia; Falco, Angela; Frati, Caterina; Gallo, Diego; Deriu, Marco A; Falvo D'Urso Labate, Giuseppe; Quaini, Federico; Audenino, Alberto; Morbiducci, Umberto

2016-01-01

A versatile bioreactor suitable for dynamic suspension cell culture under tunable shear stress conditions has been developed and preliminarily tested culturing cancer cell spheroids. By adopting simple technological solutions and avoiding rotating components, the bioreactor exploits the laminar hydrodynamics establishing within the culture chamber enabling dynamic cell suspension in an environment favourable to mass transport, under a wide range of tunable shear stress conditions. The design phase of the device has been supported by multiphysics modelling and has provided a comprehensive analysis of the operating principles of the bioreactor. Moreover, an explanatory example is herein presented with multiphysics simulations used to set the proper bioreactor operating conditions for preliminary in vitro biological tests on a human lung carcinoma cell line. The biological results demonstrate that the ultralow shear dynamic suspension provided by the device is beneficial for culturing cancer cell spheroids. In comparison to the static suspension control, dynamic cell suspension preserves morphological features, promotes intercellular connection, increases spheroid size (2.4-fold increase) and number of cycling cells (1.58-fold increase), and reduces double strand DNA damage (1.5-fold reduction). It is envisioned that the versatility of this bioreactor could allow investigation and expansion of different cell types in the future.

High density cell culture system

NASA Technical Reports Server (NTRS)

Spaulding, Glenn F. (Inventor)

1994-01-01

An annular culture vessel for growing mammalian cells is constructed in a one piece integral and annular configuration with an open end which is closed by an endcap. The culture vessel is rotatable about a horizontal axis by use of conventional roller systems commonly used in culture laboratories. The end wall of the endcap has tapered access ports to frictionally and sealingly receive the ends of hypodermic syringes. The syringes permit the introduction of fresh nutrient and withdrawal of spent nutrients. The walls are made of conventional polymeric cell culture material and are subjected to neutron bombardment to form minute gas permeable perforations in the walls.

A Multimethod Approach for Investigating Algal Toxicity of Platinum Nanoparticles.

PubMed

Sørensen, Sara N; Engelbrekt, Christian; Lützhøft, Hans-Christian H; Jiménez-Lamana, Javier; Noori, Jafar S; Alatraktchi, Fatima A; Delgado, Cristina G; Slaveykova, Vera I; Baun, Anders

2016-10-04

The ecotoxicity of platinum nanoparticles (PtNPs) widely used in for example automotive catalytic converters, is largely unknown. This study employs various characterization techniques and toxicity end points to investigate PtNP toxicity toward the green microalgae Pseudokirchneriella subcapitata and Chlamydomonas reinhardtii. Growth rate inhibition occurred in standard ISO tests (EC 50 values of 15-200 mg Pt/L), but also in a double-vial setup, separating cells from PtNPs, thus demonstrating shading as an important artifact for PtNP toxicity. Negligible membrane damage, but substantial oxidative stress was detected at 0.1-80 mg Pt/L in both algal species using flow cytometry. PtNPs caused growth rate inhibition and oxidative stress in P. subcapitata, beyond what was accounted for by dissolved Pt, indicating NP-specific toxicity of PtNPs. Overall, P. subcapitata was found to be more sensitive toward PtNPs and higher body burdens were measured in this species, possibly due to a favored binding of Pt to the polysaccharide-rich cell wall of this algal species. This study highlights the importance of using multimethod approaches in nanoecotoxicological studies to elucidate toxicity mechanisms, influence of NP-interactions with media/organisms, and ultimately to identify artifacts and appropriate end points for NP-ecotoxicity testing.

Combined algal processing: A novel integrated biorefinery process to produce algal biofuels and bioproducts

DOE PAGES

Dong, Tao; Knoshaug, Eric P.; Davis, Ryan; ...

2016-01-18

Here, the development of an integrated biorefinery process capable of producing multiple products is crucial for commercialization of microalgal biofuel production. Dilute acid pretreatment has been demonstrated as an efficient approach to utilize algal biomass more fully, by hydrolyzing microalgal carbohydrates into fermentable sugars, while making the lipids more extractable, and a protein fraction available for other products. Previously, we have shown that sugar-rich liquor could be separated from solid residue by solid-liquid separation (SLS) to produce ethanol via fermentation. However, process modeling has revealed that approximately 37% of the soluble sugars were lost in the solid cake after themore » SLS. Herein, a Combined Algal Processing (CAP) approach with a simplified configuration has been developed to improve the total energy yield. In CAP, whole algal slurry after acid pretreatment is directly used for ethanol fermentation. The ethanol and microalgal lipids can be sequentially recovered from the fermentation broth by thermal treatment and solvent extraction. Almost all the monomeric fermentable sugars can be utilized for ethanol production without compromising the lipid recovery. The techno-economic analysis (TEA) indicates that the CAP can reduce microalgal biofuel cost by $0.95 per gallon gasoline equivalent (GGE), which is a 9% reduction compared to the previous biorefinery scenario.« less

Cell culture experiments planned for the space bioreactor

NASA Technical Reports Server (NTRS)

Morrison, Dennis R.; Cross, John H.

1987-01-01

Culturing of cells in a pilot-scale bioreactor remains to be done in microgravity. An approach is presented based on several studies of cell culture systems. Previous and current cell culture research in microgravity which is specifically directed towards development of a space bioprocess is described. Cell culture experiments planned for a microgravity sciences mission are described in abstract form.

Advances in cell culture: anchorage dependence

PubMed Central

Merten, Otto-Wilhelm

2015-01-01

Anchorage-dependent cells are of great interest for various biotechnological applications. (i) They represent a formidable production means of viruses for vaccination purposes at very large scales (in 1000–6000 l reactors) using microcarriers, and in the last decade many more novel viral vaccines have been developed using this production technology. (ii) With the advent of stem cells and their use/potential use in clinics for cell therapy and regenerative medicine purposes, the development of novel culture devices and technologies for adherent cells has accelerated greatly with a view to the large-scale expansion of these cells. Presently, the really scalable systems—microcarrier/microcarrier-clump cultures using stirred-tank reactors—for the expansion of stem cells are still in their infancy. Only laboratory scale reactors of maximally 2.5 l working volume have been evaluated because thorough knowledge and basic understanding of critical issues with respect to cell expansion while retaining pluripotency and differentiation potential, and the impact of the culture environment on stem cell fate, etc., are still lacking and require further studies. This article gives an overview on critical issues common to all cell culture systems for adherent cells as well as specifics for different types of stem cells in view of small- and large-scale cell expansion and production processes. PMID:25533097

Basic Techniques in Mammalian Cell Tissue Culture.

PubMed

Phelan, Katy; May, Kristin M

2016-11-01

Cultured mammalian cells are used extensively in cell biology studies. It requires a number of special skills in order to be able to preserve the structure, function, behavior, and biology of the cells in culture. This unit describes the basic skills required to maintain and preserve cell cultures: maintaining aseptic technique, preparing media with the appropriate characteristics, passaging, freezing and storage, recovering frozen stocks, and counting viable cells. © 2016 by John Wiley & Sons, Inc. Copyright © 2016 John Wiley & Sons, Inc.

Algal toxins

USGS Publications Warehouse

Creekmore, Lynn H.

1999-01-01

Periodic blooms of algae, including true algae, dinoflagellates, and cyanobacteria or blue-green algae have been reported in marine and freshwater bodies throughout the world. Although many blooms are merely an aesthetic nuisance, some species of algae produce toxins that kill fish, shellfish, humans, livestock and wildlife. Pigmented blooms of toxinproducing marine algae are often referred to as “red tides” (Fig. 36.1). Proliferations of freshwater toxin-producing cyanobacteria are simply called “cyanobacterial blooms” or “toxic algal blooms.” Cyanobacterial blooms initially appear green and may later turn blue, sometimes forming a “scum” in the water (Fig. 36.2).Although algal blooms historically have been considered a natural phenomenon, the frequency of occurrence of harmful algae appears to have increased in recent years. Agricultural runoff and other pollutants of freshwater and marine wetlands and water bodies have resulted in increased nutrient loading of phosphorus and nitrogen, thus providing conditions favorable to the growth of potentially toxic algae. The detrimental impact of red tides and cyanobacterial blooms on wetland, shore, and pelagic species has long been suspected but not often been substantiated because information on the effects of these toxins in fish and wildlife species is lacking and diagnostic tools are limited.

Fungal-assisted algal flocculation: application in wastewater treatment and biofuel production.

PubMed

Muradov, Nazim; Taha, Mohamed; Miranda, Ana F; Wrede, Digby; Kadali, Krishna; Gujar, Amit; Stevenson, Trevor; Ball, Andrew S; Mouradov, Aidyn

2015-01-01

The microalgal-based industries are facing a number of important challenges that in turn affect their economic viability. Arguably the most important of these are associated with the high costs of harvesting and dewatering of the microalgal cells, the costs and sustainability of nutrient supplies and costly methods for large scale oil extraction. Existing harvesting technologies, which can account for up to 50% of the total cost, are not economically feasible because of either requiring too much energy or the addition of chemicals. Fungal-assisted flocculation is currently receiving increased attention because of its high harvesting efficiency. Moreover, some of fungal and microalgal strains are well known for their ability to treat wastewater, generating biomass which represents a renewable and sustainable feedstock for bioenergy production. We screened 33 fungal strains, isolated from compost, straws and soil for their lipid content and flocculation efficiencies against representatives of microalgae commercially used for biodiesel production, namely the heterotrophic freshwater microalgae Chlorella protothecoides and the marine microalgae Tetraselmis suecica. Lipid levels and composition were analyzed in fungal-algal pellets grown on media containing alternative carbon, nitrogen and phosphorus sources from wheat straw and swine wastewater, respectively. The biomass of fungal-algal pellets grown on swine wastewater was used as feedstock for the production of value-added chemicals, biogas, bio-solids and liquid petrochemicals through pyrolysis. Co-cultivation of microalgae and filamentous fungus increased total biomass production, lipid yield and wastewater bioremediation efficiency. Fungal-assisted microalgal flocculation shows significant potential for solving the major challenges facing the commercialization of microalgal biotechnology, namely (i) the efficient and cost-effective harvesting of freshwater and seawater algal strains; (ii) enhancement of total oil

Impact of harmful algal blooms on several Lake Erie drinking water treatment facilities; methodology considerations

EPA Science Inventory

The propagation of cyanbacterial cells and their toxins were investigated at seven drinking water treatment plants (DWTPs) on Lake Erie were investigated with regards to harmful algal bloom (HAB) toxin concentrations, water quality variations in treatment plant influents, and pr...

Direct conversion of algal biomass to biofuel

DOEpatents

Deng, Shuguang; Patil, Prafulla D; Gude, Veera Gnaneswar

2014-10-14

A method and system for providing direct conversion of algal biomass. Optionally, the method and system can be used to directly convert dry algal biomass to biodiesels under microwave irradiation by combining the reaction and combining steps. Alternatively, wet algae can be directly processed and converted to fatty acid methyl esters, which have the major components of biodiesels, by reacting with methanol at predetermined pressure and temperature ranges.

Pretreatment of algae-laden and manganese-containing waters by oxidation-assisted coagulation: Effects of oxidation on algal cell viability and manganese precipitation.

PubMed

Lin, Jr-Lin; Hua, Lap-Cuong; Wu, Yuting; Huang, Chihpin

2016-02-01

Preoxidation is manipulated to improve performance of algae and soluble manganese (Mn) removal by coagulation-sedimentation for water treatment plants (WTPs) when large amount of soluble Mn presents in algae-laden waters. This study aimed to investigate the effects of preoxidation on the performance of coagulation-sedimentation for the simultaneous removal of algae and soluble Mn, including ionic and complexed Mn. NaOCl, ClO2, and KMnO4 were used to pretreat such algae-laden and Mn containing waters. The variation of algal cell viability, residual cell counts, and concentrations of Mn species prior to and after coagulation-sedimentation step were investigated. Results show that NaOCl dosing was effective in reducing the viability of algae, but precipitated little Mn. ClO2 dosing had a strongest ability to lower algae viability and oxidize ionic and complexed soluble Mn, where KMnO4 dosing oxidized ionic and complexed Mn instead of reducing the viability of cells. Preoxidation by NaOCl only improved the algae removal by sedimentation, whereas most of soluble Mn still remained. On the other hand, ClO2 preoxidation substantially improved the performance of coagulation-sedimentation for simultaneous removal of algae and soluble Mn. Furthermore, KMnO4 preoxidation did improve the removal of algae by sedimentation, but left significant residual Mn in the supernatant. Images from FlowCAM showed changes in aspect ratio (AR) and transparency of algae-Mn flocs during oxidation-assisted coagulation, and indicates that an effective oxidation can improve the removal of most compact algae-Mn flocs by sedimentation. It suggests that an effective preoxidation for reducing algal cell viability and the concentration of soluble Mn is a crucial step for upgrading the performance of coagulation-sedimentation. Copyright © 2015 Elsevier Ltd. All rights reserved.

Microfluidic cardiac cell culture model (μCCCM).

PubMed

Giridharan, Guruprasad A; Nguyen, Mai-Dung; Estrada, Rosendo; Parichehreh, Vahidreza; Hamid, Tariq; Ismahil, Mohamed Ameen; Prabhu, Sumanth D; Sethu, Palaniappan

2010-09-15

Physiological heart development and cardiac function rely on the response of cardiac cells to mechanical stress during hemodynamic loading and unloading. These stresses, especially if sustained, can induce changes in cell structure, contractile function, and gene expression. Current cell culture techniques commonly fail to adequately replicate physical loading observed in the native heart. Therefore, there is a need for physiologically relevant in vitro models that recreate mechanical loading conditions seen in both normal and pathological conditions. To fulfill this need, we have developed a microfluidic cardiac cell culture model (μCCCM) that for the first time allows in vitro hemodynamic stimulation of cardiomyocytes by directly coupling cell structure and function with fluid induced loading. Cells are cultured in a small (1 cm diameter) cell culture chamber on a thin flexible silicone membrane. Integrating the cell culture chamber with a pump, collapsible pulsatile valve and an adjustable resistance element (hemostatic valve) in series allow replication of various loading conditions experienced in the heart. This paper details the design, modeling, fabrication and characterization of fluid flow, pressure and stretch generated at various frequencies to mimic hemodynamic conditions associated with the normal and failing heart. Proof-of-concept studies demonstrate successful culture of an embryonic cardiomyoblast line (H9c2 cells) and establishment of an in vivo like phenotype within this system.

Indicators: Algal Toxins (microcystin)

EPA Pesticide Factsheets

Algal toxins are toxic substances released by some types of algae (phytoplankton) when they are present in large quantities (blooms) and decay or degrade. High nutrient levels and warm temperatures often result in favorable conditions for algae blooms.

Suspension culture of pluripotent stem cells: effect of shear on stem cell fate.

PubMed

Keller, Kevin C; Rodrigues, Beatriz; zur Nieden, Nicole I

2014-01-01

Despite significant promise, the routine usage of suspension cell culture to manufacture stem cell-derived differentiated cells has progressed slowly. Suspension culture is an innovative way of either expanding or differentiating cells and sometimes both are combined into a single bioprocess. Its advantages over static 2D culturing include a homogeneous and controllable culture environment and producing a large quantity of cells in a fraction of time. This feature makes suspension cell culture ideal for use in stem cell research and eventually ideal in the large-scale production of differentiated cells for regenerative medicine. Because of their tremendous differentiation capacities and unlimited growth properties, pluripotent stem cells (PSCs) in particular are considered potential sources for future cell-replacement therapies. Currently, expansion of PSCs is accomplished in 2D, which only permits a limited amount of cell growth per culture flask before cells need to be passaged. However, before stem cells can be applied clinically, several aspects of their expansion, such as directed growth, but also differentiation, need to be better controlled. This review will summarize recent advantages in suspension culture of PSCs, while at the same time highlighting current challenges.

A strategy for urban outdoor production of high-concentration algal biomass for green biorefining.

PubMed

Lim, Chun Yong; Chen, Chia-Lung; Wang, Jing-Yuan

2013-05-01

The present study was to investigate the feasibility of carrying out effective microalgae cultivation and high-rate tertiary wastewater treatment simultaneously in a vertical sequencing batch photobioreactor with small areal footprint, suitable for sustainable urban microalgae production. For 15 consecutive days, Chlorella sorokiniana was cultivated in synthetic wastewater under various trophic conditions. A cycle of 12-h heterotrophic: 12-h mixotrophic condition produced 0.98 g l(-1) d(-1) of algal biomass in tandem with a 94.7% removal of 254.4 mg l(-1) C-acetate, a 100% removal of 84.7 mg l(-1) N-NH4 and a removal of 15.0 mg l(-1) P-PO4. The cells were harvested via cost-effective chitosan flocculation with multiple dosing (3 times) applying established chitosan:cell ratio (1:300 w/w) and pH control (6.3-6.8). Reproducible flocculation efficiencies of greater than 99% and high-concentration algal broths (>20% solids) were achieved. Copyright © 2012 Elsevier Ltd. All rights reserved.

Algal carbohydrates affect polyketide synthesis of the lichen-forming fungus Cladonia rangiferina.

PubMed

Elshobary, Mostafa E; Osman, Mohamed E; Abo-Shady, Atef M; Komatsu, Emy; Perreault, Hélène; Sorensen, John; Piercey-Normore, Michele D

2016-01-01

Lichen secondary metabolites (polyketides) are produced by the fungal partner, but the role of algal carbohydrates in polyketide biosynthesis is not clear. This study examined whether the type and concentration of algal carbohydrate explained differences in polyketide production and gene transcription by a lichen fungus (Cladonia rangiferina). The carbohydrates identified from a free-living cyanobacterium (Spirulina platensis; glucose), a lichen-forming alga (Diplosphaera chodatii; sorbitol) and the lichen alga that associates with C. rangiferina (Asterochloris sp.; ribitol) were used in each of 1%, 5% and 10% concentrations to enrich malt yeast extract media for culturing the mycobiont. Polyketides were determined by high performance liquid chromatography (HPLC), and polyketide synthase (PKS) gene transcription was measured by quantitative PCR of the ketosynthase domain of four PKS genes. The lower concentrations of carbohydrates induced the PKS gene expression where ribitol up-regulated CrPKS1 and CrPKS16 gene transcription and sorbitol up-regulated CrPKS3 and CrPKS7 gene transcription. The HPLC results revealed that lower concentrations of carbon sources increased polyketide production for three carbohydrates. One polyketide from the natural lichen thallus (fumarprotocetraric acid) also was produced by the fungal culture in ribitol supplemented media only. This study provides a better understanding of the role of the type and concentration of the carbon source in fungal polyketide biosynthesis in the lichen Cladonia rangiferina. © 2016 by The Mycological Society of America.

The evolution of chicken stem cell culture methods.

PubMed

Farzaneh, M; Attari, F; Mozdziak, P E; Khoshnam, S E

2017-12-01

1. The avian embryo is an excellent model for studying embryology and the production of pharmaceutical proteins in transgenic chickens. Furthermore, chicken stem cells have the potential for proliferation and differentiation and emerged as an attractive tool for various cell-based technologies. 2. The objective of these studies is the derivation and culture of these stem cells is the production of transgenic birds for recombinant biomaterials and vaccine manufacture, drug and cytotoxicity testing, as well as to gain insight into basic science, including cell tracking. 3. Despite similarities among the established chicken stem cell lines, fundamental differences have been reported between their culture conditions and applications. Recent conventional protocols used for expansion and culture of chicken stem cells mostly depend on feeder cells, serum-containing media and static culture. 4. Utilising chicken stem cells for generation of cell-based transgenic birds and a variety of vaccines requires large-scale cell production. However, scaling up the conventional adherent chicken stem cells is challenging and labour intensive. Development of a suspension cell culture process for chicken embryonic stem cells (cESCs), chicken primordial germ cells (PGCs) and chicken induced pluripotent stem cells (ciPSCs) will be an important advance for increasing the growth kinetics of these cells. 6. This review describes various approaches and suggestions to achieve optimal cell growth for defined chicken stem cells cultures and use in future manufacturing applications.

THE FUTURE OF HARMFUL ALGAL BLOOMS: AN EMPIRICAL APPROACH TO PREDICTING THE COMBINED IMPACTS OF RISING CO2, TEMPERATURE, AND EUTROPHICATION

EPA Science Inventory

Recent worldwide increases in harmful algal blooms (HABs) are almost certainly linked to cultural eutrophication of coastal environments. Virtually no attention has been given, however, to how other major anthropogenic impacts such as rising CO₂ and greenhouse warmi...

Mammalian Cell Culture Simplified.

ERIC Educational Resources Information Center

Moss, Robert; Solomon, Sondra

1991-01-01

A tissue culture experiment that does not require elaborate equipment and that can be used to teach sterile technique, the principles of animal cell line maintenance, and the concept of cell growth curves is described. The differences between cancerous and normal cells can be highlighted. The procedure is included. (KR)

Characterization of intracellular and extracellular saxitoxin levels in both field and cultured Alexandrium spp. samples from Sequim Bay, Washington.

PubMed

Lefebvre, Kathi A; Bill, Brian D; Erickson, Aleta; Baugh, Keri A; O'Rourke, Lohna; Costa, Pedro R; Nance, Shelly; Trainer, Vera L

2008-05-14

Traditionally, harmful algal bloom studies have primarily focused on quantifying toxin levels contained within the phytoplankton cells of interest. In the case of paralytic shellfish poisoning toxins (PSTs), intracellular toxin levels and the effects of dietary consumption of toxic cells by planktivores have been well documented. However, little information is available regarding the levels of extracellular PSTs that may leak or be released into seawater from toxic cells during blooms. In order to fully evaluate the risks of harmful algal bloom toxins in the marine food web, it is necessary to understand all potential routes of exposure. In the present study, extracellular and intracellular PST levels were measured in field seawater samples (collected weekly from June to October 2004-2007) and in Alexandrium spp. culture samples isolated from Sequim Bay, Washington. Measurable levels of intra- and extra-cellular toxins were detected in both field and culture samples via receptor binding assay (RBA) and an enzyme-linked immunosorbent assay (ELISA). Characterization of the PST toxin profile in the Sequim Bay isolates by pre-column oxidation and HPLC-fluorescence detection revealed that gonyautoxin 1 and 4 made up 65 +/- 9.7% of the total PSTs present. Collectively, these data confirm that extracellular PSTs are present during blooms of Alexandrium spp. in the Sequim Bay region.

Cultured Human Renal Cortical Cells

NASA Technical Reports Server (NTRS)

1998-01-01

During the STS-90 shuttle flight in April 1998, cultured renal cortical cells revealed new information about genes. Timothy Hammond, an investigator in NASA's microgravity biotechnology program was interested in culturing kidney tissue to study the expression of proteins useful in the treatment of kidney diseases. Protein expression is linked to the level of differentiation of the kidney cells, and Hammond had difficulty maintaining differentiated cells in vitro. Intrigued by the improvement in cell differentiation that he observed in rat renal cells cultured in NASA's rotating wall vessel (a bioreactor that simulates some aspects of microgravity) and during an experiment performed on the Russian Space Station Mir, Hammond decided to sleuth out which genes were responsible for controlling differentiation of kidney cells. To do this, he compared the gene activity of human renal cells in a variety of gravitational environments, including the microgravity of the space shuttle and the high-gravity environment of a centrifuge. Hammond found that 1,632 genes out of 10,000 analyzed changed their activity level in microgravity, more than in any of the other environments. These results have important implications for kidney research as well as for understanding the basic mechanism for controlling cell differentiation.

Recent advances in ultrasonic treatment: Challenges and field applications for controlling harmful algal blooms (HABs).

PubMed

Park, Jungsu; Church, Jared; Son, Younggyu; Kim, Keug-Tae; Lee, Woo Hyoung

2017-09-01

Algal blooms are a naturally occurring phenomenon which can occur in both freshwater and saltwater. However, due to excess nutrient loading in water bodies (e.g. agricultural runoff and industrial activities), harmful algal blooms (HABs) have become an increasing issue globally, and can even cause health effects in humans due to the release of cyanotoxins. Among currently available treatment methods, sonication has received increasing attention for algal control because of its low impact on ecosystems and the environment. The effects of ultrasound on algal cells are well understood and operating parameter such as frequency, intensity, and duration of exposure has been well studied. However, most studies have been limited to laboratory data interpretation due to complicated environmental conditions in the field. Only a few field and pilot tests in small reservoirs were reported and the applicability of ultrasound for HABs prevention and control is still under question. There is a lack of information on the upscaling of ultrasonication devices for HAB control on larger water bodies, considering field influencing factors such as rainfall, light intensity/duration, temperature, water flow, nutrients loading, and turbidity. In this review article, we address the challenges and field considerations of ultrasonic applications for controlling algal blooms. An extensive literature survey, from the fundamentals of ultrasound techniques to recent ultrasound laboratory and field studies, has been thoroughly conducted and summarized to identify future technical expectations for field applications. Case studies investigating spatial distribution of frequency and pressure during sonication are highlighted with future implications. Copyright © 2017 Elsevier B.V. All rights reserved.

Skeletal muscle satellite cells cultured in simulated microgravity

NASA Technical Reports Server (NTRS)

Molnar, Greg; Hartzell, Charles R.; Schroedl, Nancy A.; Gonda, Steve R.

1993-01-01

Satellite cells are postnatal myoblasts responsible for providing additional nuclei to growing or regenerating muscle cells. Satellite cells retain the capacity to proliferate and differentiate in vitro and therefore provide a useful model to study postnatal muscle development. Most culture systems used to study postnatal muscle development are limited by the two-dimensional (2-D) confines of the culture dish. Limiting proliferation and differentiation of satellite cells in 2-D could potentially limit cell-cell contacts important for developing the level of organization in skeletal muscle obtained in vivo. Culturing satellite cells on microcarrier beads suspended in the High-Aspect-Ratio-Vessel (HARV) designed by NASA provides a low shear, three-dimensional (3-D) environment to study muscle development. Primary cultures established from anterior tibialis muscles of growing rats (approximately 200 gm) were used for all studies and were composed of greater than 75 % satellite cells. Different inoculation densities did not affect the proliferative potential of satellite cells in the HARV. Plating efficiency, proliferation, and glucose utilization were compared between 2-D flat culture and 3-D HARV culture. Plating efficiency (cells attached - cells plated x 100) was similar between the two culture systems. Proliferation was reduced in HARV cultures and this reduction was apparent for both satellite cells and non-satellite cells. Furthermore, reduction in proliferation within the HARV could not be attributed to reduced substrate availability since glucose levels in media from HARV and 2-D cell culture were similar. Morphologically, microcarrier beads within the HARVS were joined together by cells into three-dimensional aggregates composed of greater than 10 beads/aggregate. Aggregation of beads did not occur in the absence of cells. Myotubes were often seen on individual beads or spanning the surface of two beads. In summary, proliferation and differentiation of

Got black swimming dots in your cell culture? Identification of Achromobacter as a novel cell culture contaminant

PubMed Central

Gray, Jennifer Sue; Birmingham, Janette Marie; Fenton, Jenifer Imig

2009-01-01

ARTICLE SUMMARY Cell culture model systems are utilized for their ease of use, relative inexpensiveness, and potentially limitless sample size. Reliable results cannot be obtained, however, when cultures contain contamination. This report discusses the observation and identification of mobile black specks observed in multiple cell lines. Cultures of the contamination were grown, and DNA was purified from isolated colonies. The 16S rDNA gene was PCR amplified using primers that will amplify the gene from many genera, and then sequenced. Sequencing results matched the members of the genus Achromobacter, bacteria common in the environment. Achromobacter species have been shown to be resistant to multiple antibiotics. Attempts to decontaminate the eukaryotic cell culture used multiple antibiotics at different concentrations. The contaminating Achromobacter was eventually eliminated, without permanently harming the eukaryotic cells, using a combination of the antibiotics ciprofloxacin and piperacillin. PMID:19926304

Distribution, behavior, and condition of herbivorous fishes on coral reefs track algal resources.

PubMed

Tootell, Jesse S; Steele, Mark A

2016-05-01

Herbivore distribution can impact community structure and ecosystem function. On coral reefs, herbivores are thought to play an important role in promoting coral dominance, but how they are distributed relative to algae is not well known. Here, we evaluated whether the distribution, behavior, and condition of herbivorous fishes correlated with algal resource availability at six sites in the back reef environment of Moorea, French Polynesia. Specifically, we tested the hypotheses that increased algal turf availability would coincide with (1) increased biomass, (2) altered foraging behavior, and (3) increased energy reserves of herbivorous fishes. Fish biomass and algal cover were visually estimated along underwater transects; behavior of herbivorous fishes was quantified by observations of focal individuals; fish were collected to assess their condition; and algal turf production rates were measured on standardized tiles. The best predictor of herbivorous fish biomass was algal turf production, with fish biomass increasing with algal production. Biomass of herbivorous fishes was also negatively related to sea urchin density, suggesting competition for limited resources. Regression models including both algal turf production and urchin density explained 94 % of the variation in herbivorous fish biomass among sites spread over ~20 km. Behavioral observations of the parrotfish Chlorurus sordidus revealed that foraging area increased as algal turf cover decreased. Additionally, energy reserves increased with algal turf production, but declined with herbivorous fish density, implying that algal turf is a limited resource for this species. Our findings support the hypothesis that herbivorous fishes can spatially track algal resources on coral reefs.

Effects of Nano-Titanium Dioxide on Freshwater Algal Population Dynamics

PubMed Central

Kulacki, Konrad J.; Cardinale, Bradley J.

2012-01-01

To make predictions about the possible effects of nanomaterials across environments and taxa, toxicity testing must incorporate not only a variety of organisms and endpoints, but also an understanding of the mechanisms that underlie nanoparticle toxicity. Here, we report the results of a laboratory experiment in which we examined how titanium dioxide nanoparticles impact the population dynamics and production of biomass across a range of freshwater algae. We exposed 10 of the most common species of North American freshwater pelagic algae (phytoplankton) to five increasing concentrations of n-TiO2 (ranging from controls to 300 mg n-TiO2 L−1). We then examined the effects of n-TiO2 on the population growth rates and biomass production of each algal species over a period of 25 days. On average, increasing concentrations of n-TiO2 had no significant effects on algal growth rates (p = 0.376), even though there was considerable species-specific variation in responses. In contrast, exposure to n-TiO2 tended to increase maximum biomass achieved by species in culture (p = 0.06). Results suggest that titanium dioxide nanoparticles could influence certain aspects of population growth of freshwater phytoplankton, though effects are unlikely at environmentally relevant concentrations. PMID:23071735

Distribution of heavy metals from flue gas in algal bioreactor

NASA Astrophysics Data System (ADS)

Napan, Katerine

Flue gas from coal-fired power plants is a major source of CO2 to the atmosphere. Microalgae can use this enriched form of CO2 as carbon source and in turn the biomass can be used to produce food, feed, fertilizer and biofuels. However, along with CO2, coal-based flue gas will inevitably introduce heavy metals, which have a high affinity to bind algal cells, could be toxic to the organisms and if transferred to the products could limit their uses. This study seeks to address the distribution and impact of heavy metals present in flue gas on microalgae production systems. To comprehend its effects, algae Scenedesmus obliquus was grown in batch reactors in a multimetal system. Ten heavy metals (Cu, Co, Zn, Pb, As, Se, Cr, Hg, Ni and Cd) were selected and were evaluated at four concentrations (1X, 2X, 5X and 10X). Results show that most heavy metals accumulated mainly in biomass and were found in very low concentrations in media. Hg was shown to be lost from the culture, with low amounts present in the biomass. An upper limit for As uptake was observed, suggesting its likelihood to build-up in the system during medium recycle. The As limited bioaccumulation was overcome by addition of sulfur to the algal medium. Heavy metal at 2X, 5X and 10X inhibited both growth and lipid production, while at the reference concentration both biomass and lipids yields were increased. Heavy metal concentrations in the medium and biomass were time dependent, and at the end of the cultivation most heavy metals in the supernatant solution complied with the recommendations for irrigation water, while biomass was below limits for cattle and poultry feed, fertilizer, plastic and paper. This research shows that bioremediation of CO2 and heavy metals in combination with energy production can be integrated, which is an environmentally friendly form of biotechnology.

Fiber-optic microarray for simultaneous detection of multiple harmful algal bloom species.

PubMed

Ahn, Soohyoun; Kulis, David M; Erdner, Deana L; Anderson, Donald M; Walt, David R

2006-09-01

Harmful algal blooms (HABs) are a serious threat to coastal resources, causing a variety of impacts on public health, regional economies, and ecosystems. Plankton analysis is a valuable component of many HAB monitoring and research programs, but the diversity of plankton poses a problem in discriminating toxic from nontoxic species using conventional detection methods. Here we describe a sensitive and specific sandwich hybridization assay that combines fiber-optic microarrays with oligonucleotide probes to detect and enumerate the HAB species Alexandrium fundyense, Alexandrium ostenfeldii, and Pseudo-nitzschia australis. Microarrays were prepared by loading oligonucleotide probe-coupled microspheres (diameter, 3 mum) onto the distal ends of chemically etched imaging fiber bundles. Hybridization of target rRNA from HAB cells to immobilized probes on the microspheres was visualized using Cy3-labeled secondary probes in a sandwich-type assay format. We applied these microarrays to the detection and enumeration of HAB cells in both cultured and field samples. Our study demonstrated a detection limit of approximately 5 cells for all three target organisms within 45 min, without a separate amplification step, in both sample types. We also developed a multiplexed microarray to detect the three HAB species simultaneously, which successfully detected the target organisms, alone and in combination, without cross-reactivity. Our study suggests that fiber-optic microarrays can be used for rapid and sensitive detection and potential enumeration of HAB species in the environment.

EFFECTS OF MARINE ALGAL TOXINS ON THERMOREGULATION IN MICE.

EPA Science Inventory

Hypothermia is often seen in mice and rats exposed acutely to marine algal toxins, but the mechanism of action of these toxins on thermoregulation is not well understood. Our laboratory has assessed the thermoregulatory mechanisms of two marine algal toxins, maitotoxin and brevet...

Particle Trajectories in Rotating Wall Cell Culture Devices

NASA Technical Reports Server (NTRS)

Ramachandran N.; Downey, J. P.

1999-01-01

Cell cultures are extremely important to the medical community since such cultures provide an opportunity to perform research on human tissue without the concerns inherent in experiments on individual humans. Development of cells in cultures has been found to be greatly influenced by the conditions of the culture. Much work has focused on the effect of the motions of cells in the culture relative to the solution. Recently rotating wall vessels have been used with success in achieving improved cellular cultures. Speculation and limited research have focused on the low shear environment and the ability of rotating vessels to keep cells suspended in solution rather than floating or sedimenting as the primary reasons for the improved cellular cultures using these devices. It is widely believed that the cultures obtained using a rotating wall vessel simulates to some degree the effect of microgravity on cultures. It has also been speculated that the microgravity environment may provide the ideal acceleration environment for culturing of cellular tissues due to the nearly negligible levels of sedimentation and shear possible. This work predicts particle trajectories of cells in rotating wall vessels of cylindrical and annular design consistent with the estimated properties of typical cellular cultures. Estimates of the shear encountered by cells in solution and the interactions with walls are studied. Comparisons of potential experiments in ground and microgravity environments are performed.

Embryonic Stem Cells: Isolation, Characterization and Culture

NASA Astrophysics Data System (ADS)

Amit, Michal; Itskovitz-Eldor, Joseph

Embryonic stem cells are pluripotent cells isolated from the mammalian blastocyst. Traditionally, these cells have been derived and cultured with mouse embryonic fibroblast (MEF) supportive layers, which allow their continuous growth in an undifferentiated state. However, for any future industrial or clinical application hESCs should be cultured in reproducible, defined, and xeno-free culture system, where exposure to animal pathogens is prevented. From their derivation in 1998 the methods for culturing hESCs were significantly improved. This chapter wills discuss hESC characterization and the basic methods for their derivation and maintenance.

Harmful Algal Blooms Research

EPA Science Inventory

This project represents the Agency’s first effort to unify harmful algal blooms (HABs) research that had been previously carried out in isolation within various laboratories. A unified program is the most efficient way generate useful results for the Agency’s decision...

Human cell culture in a space bioreactor

NASA Technical Reports Server (NTRS)

Morrison, Dennis R.

1988-01-01

Microgravity offers new ways of handling fluids, gases, and growing mammalian cells in efficient suspension cultures. In 1976 bioreactor engineers designed a system using a cylindrical reactor vessel in which the cells and medium are slowly mixed. The reaction chamber is interchangeable and can be used for several types of cell cultures. NASA has methodically developed unique suspension type cell and recovery apparatus culture systems for bioprocess technology experiments and production of biological products in microgravity. The first Space Bioreactor was designed for microprocessor control, no gaseous headspace, circulation and resupply of culture medium, and slow mixing in very low shear regimes. Various ground based bioreactors are being used to test reactor vessel design, on-line sensors, effects of shear, nutrient supply, and waste removal from continuous culture of human cells attached to microcarriers. The small Bioreactor is being constructed for flight experiments in the Shuttle Middeck to verify systems operation under microgravity conditions and to measure the efficiencies of mass transport, gas transfer, oxygen consumption and control of low shear stress on cells.

Responses of Pseudokirchneriella subcapitata and algal assembly to photocatalytic titanium dioxide nanoparticles

NASA Astrophysics Data System (ADS)

Metzler, David M.

Development and use of nanomaterials has increased significantly over the past decade. This trend is expected to continue for the foreseeable future, which have led some to call this new industrial revolution. One aspect of these materials that make them special is their unique properties that are different from the bulk material. These unique properties have not been investigated to determine to what extent they will impact the environment. This work was undertaken to understand how nanoparticles could impact algae. For the determination of nanoparticle toxicity, dose-response experiments were run for similar sized Al2O3, TiO2, and SiO2. Additional, a wide range of nanoparticle sizes (d1) were tested at 100 and 1000 mg/L for Al2O3, TiO 2, and SiO2. Results of different nanoparticles and similar d1 dose-response data show increased toxicity with increased surface charge of the nanoparticle. Various d1 of Al2O 3 effect the population and chlorophyll a but not lipid peroxidation. Various d1 of SiO2 and TiO2 effect the population, chlorophyll a, and lipid peroxidation. Of all TiO2 d1 tested 42 nm had the greatest effect on population, chlorophyll a, and lipid peroxidation. The effect of light intensity, algal age, and body burden was examined. The body burden was adjusted by varying the initial algal cell population while keeping the nanoparticle concentration constant. Decreased body burden decreased the effect on population. The chlorophyll a and lipid peroxidation varied with the initial decreased with decreased body burden. This trend was reversed at low body burden, the chlorophyll a and lipid peroxidation increased 3 -- 4 times greater than control values. The algal cell age was controlled by the hydraulic retention time of the pre-exposure continuously stirred tank reactors. As the age of the algae increased the effect of population increased. At algae age great then 10 days the effect on population reminded constant. Titanium dioxide effect on chlorophyll a

Cell division in Escherichia coli cultures monitored at single cell resolution

PubMed Central

Roostalu, Johanna; Jõers, Arvi; Luidalepp, Hannes; Kaldalu, Niilo; Tenson, Tanel

2008-01-01

Background A fundamental characteristic of cells is the ability to divide. To date, most parameters of bacterial cultures, including cell division, have been measured as cell population averages, assuming that all bacteria divide at a uniform rate. Results We monitored the division of individual cells in Escherichia coli cultures during different growth phases. Our experiments are based on the dilution of green fluorescent protein (GFP) upon cell division, monitored by flow cytometry. The results show that the vast majority of E. coli cells in exponentially growing cultures divided uniformly. In cultures that had been in stationary phase up to four days, no cell division was observed. However, upon dilution of stationary phase culture into fresh medium, two subpopulations of cells emerged: one that started dividing and another that did not. These populations were detectable by GFP dilution and displayed different side scatter parameters in flow cytometry. Further analysis showed that bacteria in the non-growing subpopulation were not dead, neither was the difference in growth capacity reducible to differences in stationary phase-specific gene expression since we observed uniform expression of several stress-related promoters. The presence of non-growing persisters, temporarily dormant bacteria that are tolerant to antibiotics, has previously been described within growing bacterial populations. Using the GFP dilution method combined with cell sorting, we showed that ampicillin lyses growing bacteria while non-growing bacteria retain viability and that some of them restart growth after the ampicillin is removed. Thus, our method enables persisters to be monitored even in liquid cultures of wild type strains in which persister formation has low frequency. Conclusion In principle, the approaches developed here could be used to detect differences in cell division in response to different environmental conditions and in cultures of unicellular organisms other than E

COMPARISON OF LARGE RIVER SAMPLING METHODS ON ALGAL METRICS

EPA Science Inventory

We compared the results of four methods used to assess the algal communities at 60 sites distributed among four rivers. Based on Principle Component Analysis of physical habitat data collected concomitantly with the algal data, sites were separated into those with a mean thalweg...

Good Cell Culture Practice for stem cells and stem-cell-derived models.

PubMed

Pamies, David; Bal-Price, Anna; Simeonov, Anton; Tagle, Danilo; Allen, Dave; Gerhold, David; Yin, Dezhong; Pistollato, Francesca; Inutsuka, Takashi; Sullivan, Kristie; Stacey, Glyn; Salem, Harry; Leist, Marcel; Daneshian, Mardas; Vemuri, Mohan C; McFarland, Richard; Coecke, Sandra; Fitzpatrick, Suzanne C; Lakshmipathy, Uma; Mack, Amanda; Wang, Wen Bo; Yamazaki, Daiju; Sekino, Yuko; Kanda, Yasunari; Smirnova, Lena; Hartung, Thomas

2017-01-01

The first guidance on Good Cell Culture Practice (GCCP) dates back to 2005. This document expands this to include aspects of quality assurance for in vitro cell culture focusing on the increasingly diverse cell types and culture formats used in research, product development, testing and manufacture of biotechnology products and cell-based medicines. It provides a set of basic principles of best practice that can be used in training new personnel, reviewing and improving local procedures, and helping to assure standard practices and conditions for the comparison of data between laboratories and experimentation performed at different times. This includes recommendations for the documentation and reporting of culture conditions. It is intended as guidance to facilitate the generation of reliable data from cell culture systems, and is not intended to conflict with local or higher level legislation or regulatory requirements. It may not be possible to meet all recommendations in this guidance for practical, legal or other reasons. However, when it is necessary to divert from the principles of GCCP, the risk of decreasing the quality of work and the safety of laboratory staff should be addressed and any conclusions or alternative approaches justified. This workshop report is considered a first step toward a revised GCCP 2.0.

21 CFR 864.2280 - Cultured animal and human cells.

Code of Federal Regulations, 2010 CFR

2010-04-01

... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Cultured animal and human cells. 864.2280 Section... (CONTINUED) MEDICAL DEVICES HEMATOLOGY AND PATHOLOGY DEVICES Cell And Tissue Culture Products § 864.2280 Cultured animal and human cells. (a) Identification. Cultured animal and human cells are in vitro...

Long-term maintenance of human induced pluripotent stem cells by automated cell culture system.

PubMed

Konagaya, Shuhei; Ando, Takeshi; Yamauchi, Toshiaki; Suemori, Hirofumi; Iwata, Hiroo

2015-11-17

Pluripotent stem cells, such as embryonic stem cells and induced pluripotent stem (iPS) cells, are regarded as new sources for cell replacement therapy. These cells can unlimitedly expand under undifferentiated conditions and be differentiated into multiple cell types. Automated culture systems enable the large-scale production of cells. In addition to reducing the time and effort of researchers, an automated culture system improves the reproducibility of cell cultures. In the present study, we newly designed a fully automated cell culture system for human iPS maintenance. Using an automated culture system, hiPS cells maintained their undifferentiated state for 60 days. Automatically prepared hiPS cells had a potency of differentiation into three germ layer cells including dopaminergic neurons and pancreatic cells.

High-Throughput Cancer Cell Sphere Formation for 3D Cell Culture.

PubMed

Chen, Yu-Chih; Yoon, Euisik

2017-01-01

Three-dimensional (3D) cell culture is critical in studying cancer pathology and drug response. Though 3D cancer sphere culture can be performed in low-adherent dishes or well plates, the unregulated cell aggregation may skew the results. On contrary, microfluidic 3D culture can allow precise control of cell microenvironments, and provide higher throughput by orders of magnitude. In this chapter, we will look into engineering innovations in a microfluidic platform for high-throughput cancer cell sphere formation and review the implementation methods in detail.

Algal MIPs, high diversity and conserved motifs

PubMed Central

2011-01-01

Background Major intrinsic proteins (MIPs) also named aquaporins form channels facilitating the passive transport of water and other small polar molecules across membranes. MIPs are particularly abundant and diverse in terrestrial plants but little is known about their evolutionary history. In an attempt to investigate the origin of the plant MIP subfamilies, genomes of chlorophyte algae, the sister group of charophyte algae and land plants, were searched for MIP encoding genes. Results A total of 22 MIPs were identified in the nine analysed genomes and phylogenetic analyses classified them into seven subfamilies. Two of these, Plasma membrane Intrinsic Proteins (PIPs) and GlpF-like Intrinsic Proteins (GIPs), are also present in land plants and divergence dating support a common origin of these algal and land plant MIPs, predating the evolution of terrestrial plants. The subfamilies unique to algae were named MIPA to MIPE to facilitate the use of a common nomenclature for plant MIPs reflecting phylogenetically stable groups. All of the investigated genomes contained at least one MIP gene but only a few species encoded MIPs belonging to more than one subfamily. Conclusions Our results suggest that at least two of the seven subfamilies found in land plants were present already in an algal ancestor. The total variation of MIPs and the number of different subfamilies in chlorophyte algae is likely to be even higher than that found in land plants. Our analyses indicate that genetic exchanges between several of the algal subfamilies have occurred. The PIP1 and PIP2 groups and the Ca2+ gating appear to be specific to land plants whereas the pH gating is a more ancient characteristic shared by all PIPs. Further studies are needed to discern the function of the algal specific subfamilies MIPA-E and to fully understand the evolutionary relationship of algal and terrestrial plant MIPs. PMID:21510875

Algal MIPs, high diversity and conserved motifs.

PubMed

Anderberg, Hanna I; Danielson, Jonas Å H; Johanson, Urban

2011-04-21

Major intrinsic proteins (MIPs) also named aquaporins form channels facilitating the passive transport of water and other small polar molecules across membranes. MIPs are particularly abundant and diverse in terrestrial plants but little is known about their evolutionary history. In an attempt to investigate the origin of the plant MIP subfamilies, genomes of chlorophyte algae, the sister group of charophyte algae and land plants, were searched for MIP encoding genes. A total of 22 MIPs were identified in the nine analysed genomes and phylogenetic analyses classified them into seven subfamilies. Two of these, Plasma membrane Intrinsic Proteins (PIPs) and GlpF-like Intrinsic Proteins (GIPs), are also present in land plants and divergence dating support a common origin of these algal and land plant MIPs, predating the evolution of terrestrial plants. The subfamilies unique to algae were named MIPA to MIPE to facilitate the use of a common nomenclature for plant MIPs reflecting phylogenetically stable groups. All of the investigated genomes contained at least one MIP gene but only a few species encoded MIPs belonging to more than one subfamily. Our results suggest that at least two of the seven subfamilies found in land plants were present already in an algal ancestor. The total variation of MIPs and the number of different subfamilies in chlorophyte algae is likely to be even higher than that found in land plants. Our analyses indicate that genetic exchanges between several of the algal subfamilies have occurred. The PIP1 and PIP2 groups and the Ca2+ gating appear to be specific to land plants whereas the pH gating is a more ancient characteristic shared by all PIPs. Further studies are needed to discern the function of the algal specific subfamilies MIPA-E and to fully understand the evolutionary relationship of algal and terrestrial plant MIPs.

Highly-efficient enzymatic conversion of crude algal oils into biodiesel.

PubMed

Wang, Yao; Liu, Jin; Gerken, Henri; Zhang, Chengwu; Hu, Qiang; Li, Yantao

2014-11-01

Energy-intensive chemical conversion of crude algal oils into biodiesel is a major barrier for cost-effective algal biofuel production. To overcome this problem, we developed an enzyme-based platform for conversion of crude algal oils into fatty acid methyl esters. Crude algal oils were extracted from the oleaginous microalga Nannochloropsis oceanica IMET1 and converted by an immobilized lipase from Candida antarctica. The effects of different acyl acceptors, t-butanol as a co-solvent, oil to t-butanol ratio, oil to methanol ratio, temperature and reaction time on biodiesel conversion efficiency were studied. The conversion efficiency reached 99.1% when the conversion conditions were optimized, i.e., an oil to t-butanol weight ratio of 1:1, an oil to methanol molar ratio of 1:12, and a reaction time of 4h at 25°C. The enzymatic conversion process developed in this study may hold a promise for low energy consumption, low wastewater-discharge biochemical conversion of algal feedstocks into biofuels. Published by Elsevier Ltd.

Morphology of primary human venous endothelial cell cultures before and after culture medium exchange.

PubMed

Krüger-Genge, A; Fuhrmann, R; Jung, F; Franke, R P

2015-01-01

The evaluation of the interaction of human, venous endothelial cells (HUVEC) with body foreign materials on the cellular level cannot be performed in vivo, but is investigated in vitro under standard culture conditions. To maintain the vitality, proliferation and morphology of HUVEC seeded on body foreign substrates over days, the cell culture medium is usually exchanged every second day. It is well known, that alterations in the microenvironment of cells bear the risk of influencing cell morphology and function. In the current study the influence of cell culture medium exchange on HUVEC cytoskeletal microfilament structure and function was investigated. HUVEC in the third passage were seeded on extracellular matrix (ECM) - which was secreted from bovine corneal endothelial cells on glass- until functional confluence was reached. The experiment started 11 days after HUVEC seeding with an exchange of the cell culture medium followed by a staining of the actin microfilaments with phalloidin-rhodamin 1.5 and 5 minutes after medium exchange. The microfilaments were documented by use of an Olympus microscope (IMT-2) equipped with a UV lamp and online connected to a TV chain (Sony XC 50 ST/monochrome) implying an OPTIMAS - Image analysis system. Prostacyclin was analysed in the cell culture supernatant. 1.5 min after culture medium exchange in the functionally confluent cultures a slight disturbance of the actin microfilament structure with a broadening of the marginal filament band, a partial disconnection of cell-cell contacts and the appearance of intercellular fenestrations were observed. 5 minutes after medium exchange a redevelopment of the slightly disturbed microfilament structure with a condensation and narrowing of the marginal filament band was seen. 12 h later a further consolidation of the microfilament structure occurred. In addition, a perturbation of the cultured HUVEC occurred after cell culture medium exchange. The prostacyclin concentration in the

Biosynthesis of 14C-phytoene from tomato cell suspension cultures (Lycopersicon esculentum) for utilization in prostate cancer cell culture studies.

PubMed

Campbell, Jessica K; Rogers, Randy B; Lila, Mary Ann; Erdman, John W

2006-02-08

This work describes the development and utilization of a plant cell culture production approach to biosynthesize and radiolabel phytoene and phytofluene for prostate cancer cell culture studies. The herbicide norflurazon was added to established cell suspension cultures of tomato (Lycopersicon esculentum cv. VFNT cherry), to induce the biosynthesis and accumulation of the lycopene precursors, phytoene and phytofluene, in their natural isomeric forms (15-cis-phytoene and two cis-phytofluene isomers). Norflurazon concentrations, solvent carrier type and concentration, and duration of culture exposure to norflurazon were screened to optimize phytoene and phytofluene synthesis. Maximum yields of both phytoene and phytofluene were achieved after 7 days of treatment with 0.03 mg norflurazon/40 mL fresh medium, provided in 0.07% solvent carrier. Introduction of 14C-sucrose to the tomato cell culture medium enabled the production of 14C-labeled phytoene for subsequent prostate tumor cell uptake studies. In DU 145 prostate tumor cells, it was determined that 15-cis-phytoene and an oxidized product of phytoene were taken up and partially metabolized by the cells. The ability to biosynthesize, radiolabel, and isolate these carotenoids from tomato cell cultures is a novel, valuable methodology for further in vitro and in vivo investigations into the roles of phytoene and phytofluene in cancer chemoprevention.

Advantages and challenges of microfluidic cell culture in polydimethylsiloxane devices.

PubMed

Halldorsson, Skarphedinn; Lucumi, Edinson; Gómez-Sjöberg, Rafael; Fleming, Ronan M T

2015-01-15

Culture of cells using various microfluidic devices is becoming more common within experimental cell biology. At the same time, a technological radiation of microfluidic cell culture device designs is currently in progress. Ultimately, the utility of microfluidic cell culture will be determined by its capacity to permit new insights into cellular function. Especially insights that would otherwise be difficult or impossible to obtain with macroscopic cell culture in traditional polystyrene dishes, flasks or well-plates. Many decades of heuristic optimization have gone into perfecting conventional cell culture devices and protocols. In comparison, even for the most commonly used microfluidic cell culture devices, such as those fabricated from polydimethylsiloxane (PDMS), collective understanding of the differences in cellular behavior between microfluidic and macroscopic culture is still developing. Moving in vitro culture from macroscopic culture to PDMS based devices can come with unforeseen challenges. Changes in device material, surface coating, cell number per unit surface area or per unit media volume may all affect the outcome of otherwise standard protocols. In this review, we outline some of the advantages and challenges that may accompany a transition from macroscopic to microfluidic cell culture. We focus on decisive factors that distinguish macroscopic from microfluidic cell culture to encourage a reconsideration of how macroscopic cell culture principles might apply to microfluidic cell culture. Copyright © 2014 The Authors. Published by Elsevier B.V. All rights reserved.

Strain, biochemistry, and cultivation-dependent measurement variability of algal biomass composition.

PubMed

Laurens, Lieve M L; Van Wychen, Stefanie; McAllister, Jordan P; Arrowsmith, Sarah; Dempster, Thomas A; McGowen, John; Pienkos, Philip T

2014-05-01

Accurate compositional analysis in biofuel feedstocks is imperative; the yields of individual components can define the economics of an entire process. In the nascent industry of algal biofuels and bioproducts, analytical methods that have been deemed acceptable for decades are suddenly critical for commercialization. We tackled the question of how the strain and biochemical makeup of algal cells affect chemical measurements. We selected a set of six procedures (two each for lipids, protein, and carbohydrates): three rapid fingerprinting methods and three advanced chromatography-based methods. All methods were used to measure the composition of 100 samples from three strains: Scenedesmus sp., Chlorella sp., and Nannochloropsis sp. The data presented point not only to species-specific discrepancies but also to cell biochemistry-related discrepancies. There are cases where two respective methods agree but the differences are often significant with over- or underestimation of up to 90%, likely due to chemical interferences with the rapid spectrophotometric measurements. We provide background on the chemistry of interfering reactions for the fingerprinting methods and conclude that for accurate compositional analysis of algae and process and mass balance closure, emphasis should be placed on unambiguous characterization using methods where individual components are measured independently. Copyright © 2014 Elsevier Inc. All rights reserved.

Growth and acid production of Lactobacillus delbrueckii ssp. bulgaricus ATCC 11842 in the fermentation of algal carcass.

PubMed

Li, C; Zhang, G F; Mao, X; Wang, J Y; Duan, C Y; Wang, Z J; Liu, L B

2016-06-01

Algal carcass is a low-value byproduct of algae after its conversion to biodiesel. Dried algal carcass is rich in protein, carbohydrate, and multiple amino acids, and it is typically well suited for growth and acid production of lactic acid bacteria. In this study, Lactobacillus delbrueckii ssp. bulgaricus ATCC 11842 was used to ferment different algal carcass media (ACM), including 2% ACM, 2% ACM with 1.9% glucose (ACM-G), and 2% ACM with 1.9% glucose and 2g/L amino acid mixture (ACM-GA). Concentrations of organic acids (lactic acid and acetic acid), acetyl-CoA, and ATP were analyzed by HPLC, and activities of lactate dehydrogenase (LDH), acetokinase (ACK), pyruvate kinase (PK), and phosphofructokinase (PFK) were determined by using a chemical approach. The growth of L. bulgaricus cells in ACM-GA was close to that in the control medium (de Man, Rogosa, and Sharpe). Lactic acid and acetic acid contents were greatly reduced when L. bulgaricus cells were grown in ACM compared with the control medium. Acetyl-CoA content varied with organic acid content and was increased in cells grown in different ACM compared with the control medium. The ATP content of L. bulgaricus cells in ACM was reduced compared with that of cells grown in the control medium. Activities of PFK and ACK of L. bulgaricus cells grown in ACM were higher and those of PK and LDH were lower compared with the control. Thus, ACM rich in nutrients may serve as an excellent substrate for growth by lactic acid bacteria, and addition of appropriate amounts of glucose and amino acids can improve growth and acid production. Copyright © 2016 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

A Methylobacterium-like organism from algal crusts covering silicone rubber electric insulators in Africa.

PubMed

Zarnowski, R; Felske, A; Ellis, R J; Geuns, J M C; Pietr, S J

2002-01-01

The primary goals of this study were to isolate, identify and characterize culturable bacteria living in a close association with microalgae within green crusts covering silicone rubber electric insulators in Tanzania. Twenty-four bacterial colonies were isolated from an Apatococcus crust. Characterization by statistical analyses of total cellular protein profiles demonstrated that they were highly similar to one another. Final identification was achieved using 16S rDNA sequencing and fatty acid methyl ester profiling. These analyses revealed the presence of microbes with high similarity to Methylobacterium radiotolerans. The selected isolate, A1, displayed strong inhibitory activity against Rhizoctonia solani and was found to be resistant to relatively high concentrations of zinc in the growth medium. This study revealed the presence of M. radiotolerans bacteria in a novel environment--within algal crusts formed on electrical insulators in Africa. Moreover, this bacterium was found to be a predominant culturable species within those complex algal-microbial associations. The isolate also shared some traits of biotechnological importance with other members of the Methylobacterium genus. The data presented provide a valuable contribution concerning the formation and function of associations between green microalgae and bacteria. This study also provides some information about the utility of bacteria from the genus Methylobacterium in biotechnological applications, such as biocontrol of rhizoctoniosis and bioremediation of heavy metal-contaminated soils.

Introducing Mammalian Cell Culture and Cell Viability Techniques in the Undergraduate Biology Laboratory.

PubMed

Bowey-Dellinger, Kristen; Dixon, Luke; Ackerman, Kristin; Vigueira, Cynthia; Suh, Yewseok K; Lyda, Todd; Sapp, Kelli; Grider, Michael; Crater, Dinene; Russell, Travis; Elias, Michael; Coffield, V McNeil; Segarra, Verónica A

2017-01-01

Undergraduate students learn about mammalian cell culture applications in introductory biology courses. However, laboratory modules are rarely designed to provide hands-on experience with mammalian cells or teach cell culture techniques, such as trypsinization and cell counting. Students are more likely to learn about cell culture using bacteria or yeast, as they are typically easier to grow, culture, and manipulate given the equipment, tools, and environment of most undergraduate biology laboratories. In contrast, the utilization of mammalian cells requires a dedicated biological safety cabinet and rigorous antiseptic techniques. For this reason, we have devised a laboratory module and method herein that familiarizes students with common cell culture procedures, without the use of a sterile hood or large cell culture facility. Students design and perform a time-efficient inquiry-based cell viability experiment using HeLa cells and tools that are readily available in an undergraduate biology laboratory. Students will become familiar with common techniques such as trypsinizing cells, cell counting with a hemocytometer, performing serial dilutions, and determining cell viability using trypan blue dye. Additionally, students will work with graphing software to analyze their data and think critically about the mechanism of death on a cellular level. Two different adaptations of this inquiry-based lab are presented-one for non-biology majors and one for biology majors. Overall, these laboratories aim to expose students to mammalian cell culture and basic techniques and help them to conceptualize their application in scientific research.

Multizone Paper Platform for 3D Cell Cultures

PubMed Central

Derda, Ratmir; Hong, Estrella; Mwangi, Martin; Mammoto, Akiko; Ingber, Donald E.; Whitesides, George M.

2011-01-01

In vitro 3D culture is an important model for tissues in vivo. Cells in different locations of 3D tissues are physiologically different, because they are exposed to different concentrations of oxygen, nutrients, and signaling molecules, and to other environmental factors (temperature, mechanical stress, etc). The majority of high-throughput assays based on 3D cultures, however, can only detect the average behavior of cells in the whole 3D construct. Isolation of cells from specific regions of 3D cultures is possible, but relies on low-throughput techniques such as tissue sectioning and micromanipulation. Based on a procedure reported previously (“cells-in-gels-in-paper” or CiGiP), this paper describes a simple method for culture of arrays of thin planar sections of tissues, either alone or stacked to create more complex 3D tissue structures. This procedure starts with sheets of paper patterned with hydrophobic regions that form 96 hydrophilic zones. Serial spotting of cells suspended in extracellular matrix (ECM) gel onto the patterned paper creates an array of 200 micron-thick slabs of ECM gel (supported mechanically by cellulose fibers) containing cells. Stacking the sheets with zones aligned on top of one another assembles 96 3D multilayer constructs. De-stacking the layers of the 3D culture, by peeling apart the sheets of paper, “sections” all 96 cultures at once. It is, thus, simple to isolate 200-micron-thick cell-containing slabs from each 3D culture in the 96-zone array. Because the 3D cultures are assembled from multiple layers, the number of cells plated initially in each layer determines the spatial distribution of cells in the stacked 3D cultures. This capability made it possible to compare the growth of 3D tumor models of different spatial composition, and to examine the migration of cells in these structures. PMID:21573103

Beach-goer behavior during a retrospectively detected algal ...

EPA Pesticide Factsheets

Algal blooms occur among nutrient rich, warm surface waters and may adversely impact recreational beaches. During July – September 2003, a prospective study of beachgoers was conducted on weekends at a public beach on a Great Lake in the United States. We measured each beachgoer’s activity at the start and end of their beach visit and the environmental factors: water and air temperature, wind speed and wave height at the study site each day. At the time, there was no notification of algal blooms; we retrospectively evaluated the presence of algal blooms using MERIS data from the Envisat-1 satellite. A total of 2840 people participated in the study over 16 study days. The majority (55%) were female, and 751 (26%) were < 18 years of age. An algal bloom was detected retrospectively by remotely sensed satellite imagery during August 16 – 24. This peak bloom period (PB) included 4 study days. During PB study days, more study participants 226/742 (31%) reported body contact with the water compared to contact 531/2098 (25%) on non-peak days. During the 4 PB days, of the environmental factors, only mean water temperature was significantly different, 250 C vs. 230 C (p<0.05) from other days.These results suggest that beachgoer body contact with water was not deterred by the presence of an algal bloom, and that interventions to actively discourage water contact during a bloom are needed to reduce exposure to blooms. This is an abstract of a proposed presentation and

Cell culture's spider silk road.

PubMed

Perkel, Jeffrey

2014-06-01

A number of synthetic and natural materials have been tried in cell culture and tissue engineering applications in recent years. Now Jeffrey Perkel takes a look at one new culture component that might surprise you-spider silk.

Cell culture techniques in honey bee research

USDA-ARS?s Scientific Manuscript database

Cell culture techniques are indispensable in most if not all life science disciplines to date. Wherever cell culture models are lacking scientific development is hampered. Unfortunately this has been and still is the case in honey bee research because permanent honey bee cell lines have not yet been...

Algal taxonomy: a road to nowhere?

PubMed

De Clerck, Olivier; Guiry, Michael D; Leliaert, Frederik; Samyn, Yves; Verbruggen, Heroen

2013-04-01

The widespread view of taxonomy as an essentially retrogressive and outmoded science unable to cope with the current biodiversity crisis stimulated us to analyze the current status of cataloguing global algal diversity. Contrary to this largely pessimistic belief, species description rates of algae through time and trends in the number of active taxonomists, as revealed by the web resource AlgaeBase, show a much more positive picture. More species than ever before are being described by a large community of algal taxonomists. The lack of any decline in the rate at which new species and genera are described, however, is indicative of the large proportion of undiscovered diversity and bears heavily on any prediction of global algal species diversity and the time needed to catalogue it. The saturation of accumulation curves of higher taxa (family, order, and classes) on the other hand suggest that at these taxonomic levels most diversity has been discovered. This reasonably positive picture does not imply that algal taxonomy does not face serious challenges in the near future. The observed levels of cryptic diversity in algae, combined with the shift in methods used to characterize them, have resulted in a rampant uncertainty about the status of many older species. As a consequence, there is a tendency in phycology to move gradually away from traditional names to a more informal system whereby clade-, specimen- or strain-based identifiers are used to communicate biological information. Whether these informal names for species-level clades represent a temporary situation stimulated by the lag between species discovery and formal description, or an incipient alternative or parallel taxonomy, will be largely determined by how well we manage to integrate historical collections into modern taxonomic research. Additionally, there is a pressing need for a consensus about the organizational framework to manage the information about algal species names. An eventual strategy

Stress-Survival Gene Identification From an Acid Mine Drainage Algal Mat Community

NASA Astrophysics Data System (ADS)

Urbina-Navarrete, J.; Fujishima, K.; Paulino-Lima, I. G.; Rothschild-Mancinelli, B.; Rothschild, L. J.

2014-12-01

Microbial communities from acid mine drainage environments are exposed to multiple stressors to include low pH, high dissolved metal loads, seasonal freezing, and desiccation. The microbial and algal communities that inhabit these niche environments have evolved strategies that allow for their ecological success. Metagenomic analyses are useful in identifying species diversity, however they do not elucidate the mechanisms that allow for the resilience of a community under these extreme conditions. Many known or predicted genes encode for protein products that are unknown, or similarly, many proteins cannot be traced to their gene of origin. This investigation seeks to identify genes that are active in an algal consortium during stress from living in an acid mine drainage environment. Our approach involves using the entire community transcriptome for a functional screen in an Escherichia coli host. This approach directly targets the genes involved in survival, without need for characterizing the members of the consortium.The consortium was harvested and stressed with conditions similar to the native environment it was collected from. Exposure to low pH (< 3.2), high metal load, desiccation, and deep freeze resulted in the expression of stress-induced genes that were transcribed into messenger RNA (mRNA). These mRNA transcripts were harvested to build complementary DNA (cDNA) libraries in E. coli. The transformed E. coli were exposed to the same stressors as the original algal consortium to select for surviving cells. Successful cells incorporated the transcripts that encode survival mechanisms, thus allowing for selection and identification of the gene(s) involved. Initial selection screens for freeze and desiccation tolerance have yielded E. coli that are 1 order of magnitude more resistant to freezing (0.01% survival of control with no transcript, 0.2% survival of E. coli with transcript) and 3 orders of magnitude more resistant to desiccation (0.005% survival of

The effect of TRAIL molecule on cell viability in in vitro beta cell culture.

PubMed

Tekmen, I; Ozyurt, D; Pekçetin, C; Buldan, Z

2007-06-01

Insulin-dependent diabetes mellitus (IDDM) is an organ-specific autoimmune disorder triggered by autoreactive T cells directed to pancreas beta-cell antigens. In this disorder, more than 90% of beta cells are destroyed. Cell death may be mediated via soluble or membrane-bound cell death ligands. One of these ligands may be tumour necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL), a member of the TNF-alpha superfamily. In the present study, we examined whether TRAIL had cytotoxic effects on adult rat pancreas beta cell cultures and INS1-E rat insulinoma cell line cultures or not. In this study, cell destruction models were built with TRAIL concentrations of 10, 100 and 1000 ng. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) test was used for evaluating cell viability. It was detected that cell cultures with TRAIL added showed no differences statistically when compared with control cultures containing no toxic additions. These results showed that TRAIL did not have significant cytotoxic effects on pancreas beta cell culture and INS-1E rat insulinoma cell line cultures. Detection of the expression of TRAIL receptors and natural apoptosis inhibitor proteins will be favourable to investigate the resistance mechanisms to TRAIL-induced cell death in this cell culture system.

Cell sources for in vitro human liver cell culture models

PubMed Central

Freyer, Nora; Damm, Georg; Seehofer, Daniel; Knöspel, Fanny

2016-01-01

In vitro liver cell culture models are gaining increasing importance in pharmacological and toxicological research. The source of cells used is critical for the relevance and the predictive value of such models. Primary human hepatocytes (PHH) are currently considered to be the gold standard for hepatic in vitro culture models, since they directly reflect the specific metabolism and functionality of the human liver; however, the scarcity and difficult logistics of PHH have driven researchers to explore alternative cell sources, including liver cell lines and pluripotent stem cells. Liver cell lines generated from hepatomas or by genetic manipulation are widely used due to their good availability, but they are generally altered in certain metabolic functions. For the past few years, adult and pluripotent stem cells have been attracting increasing attention, due their ability to proliferate and to differentiate into hepatocyte-like cells in vitro. However, controlling the differentiation of these cells is still a challenge. This review gives an overview of the major human cell sources under investigation for in vitro liver cell culture models, including primary human liver cells, liver cell lines, and stem cells. The promises and challenges of different cell types are discussed with a focus on the complex 2D and 3D culture approaches under investigation for improving liver cell functionality in vitro. Finally, the specific application options of individual cell sources in pharmacological research or disease modeling are described. PMID:27385595

Sensing Cell-Culture Assays with Low-Cost Circuitry.

PubMed

Pérez, Pablo; Huertas, Gloria; Maldonado-Jacobi, Andrés; Martín, María; Serrano, Juan A; Olmo, Alberto; Daza, Paula; Yúfera, Alberto

2018-06-11

An alternative approach for cell-culture end-point protocols is proposed herein. This new technique is suitable for real-time remote sensing. It is based on Electrical Cell-substrate Impedance Spectroscopy (ECIS) and employs the Oscillation-Based Test (OBT) method. Simple and straightforward circuit blocks form the basis of the proposed measurement system. Oscillation parameters - frequency and amplitude - constitute the outcome, directly correlated with the culture status. A user can remotely track the evolution of cell cultures in real time over the complete experiment through a web tool continuously displaying the acquired data. Experiments carried out with commercial electrodes and a well-established cell line (AA8) are described, obtaining the cell number in real time from growth assays. The electrodes have been electrically characterized along the design flow in order to predict the system performance and the sensitivity curves. Curves for 1-week cell growth are reported. The obtained experimental results validate the proposed OBT for cell-culture characterization. Furthermore, the proposed electrode model provides a good approximation for the cell number and the time evolution of the studied cultures.

Cell Culture as an Alternative in Education.

ERIC Educational Resources Information Center

Nardone, Roland M.

1990-01-01

Programs that are intended to inform and provide "hands-on" experience for students and to facilitate the introduction of cell culture-based laboratory exercises into the high school and college laboratory are examined. The components of the CellServ Program and the Cell Culture Toxicology Training Programs are described. (KR)

The usefulness of three-dimensional cell culture in induction of cancer stem cells from esophageal squamous cell carcinoma cell lines

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

Fujiwara, Daisuke; Kato, Kazunori, E-mail: kzkatou@juntendo.ac.jp; Department of Atopy Research Center, Juntendo University School of Medicine, 2-1-1 Hongo, Bunkyo-ku, Tokyo 113-8421

2013-05-17

Highlights: •Spheroids were created from esophageal carcinoma cells using NanoCulture® Plates. •The proportion of strongly ALDH-positive cells increased in 3-D culture. •Expression of cancer stem cell-related genes was enhanced in 3-D culture. •CA-9 expression was enhanced, suggesting hypoxia had been induced in 3-D culture. •Drug resistance was increased. 3-D culture is useful for inducing cancer stem cells. -- Abstract: In recent years, research on resistance to chemotherapy and radiotherapy in cancer treatment has come under the spotlight, and researchers have also begun investigating the relationship between resistance and cancer stem cells. Cancer stem cells are assumed to be present inmore » esophageal cancer, but experimental methods for identification and culture of these cells have not yet been established. To solve this problem, we created spheroids using a NanoCulture® Plate (NCP) for 3-dimensional (3-D) cell culture, which was designed as a means for experimentally reproducing the 3-D structures found in the body. We investigated the potential for induction of cancer stem cells from esophageal cancer cells. Using flow cytometry we analyzed the expression of surface antigen markers CD44, CD133, CD338 (ABCG2), CD318 (CDCP1), and CD326 (EpCAM), which are known cancer stem cell markers. None of these surface antigen markers showed enhanced expression in 3-D cultured cells. We then analyzed aldehyde dehydrogenase (ALDH) enzymatic activity using the ALDEFLUOR reagent, which can identify immature cells such as stem cells and precursor cells. 3-D-cultured cells were strongly positive for ALDH enzyme activity. We also analyzed the expression of the stem cell-related genes Sox-2, Nanog, Oct3/4, and Lin28 using RT-PCR. Expression of Sox-2, Nanog, and Lin28 was enhanced. Analysis of expression of the hypoxic surface antigen marker carbonic anhydrase-9 (CA-9), which is an indicator of cancer stem cell induction and maintenance, revealed that CA-9

Recycled de-Oiled Algal Biomass Extract as a Feedstock for Boosting Biodiesel Production from Chlorella minutissima.

PubMed

Arora, Neha; Patel, Alok; Pruthi, Parul A; Pruthi, Vikas

2016-12-01

The investigation for the first time assesses the efficacy of recycled de-oiled algal biomass extract (DABE) as a cultivation media to boost lipid productivity in Chlorella minutissima and its comparison with Bold's basal media (BBM) used as control. Presence of organic carbon (3.8 ± 0.8 g/l) in recycled DABE resulted in rapid growth with twofold increase in biomass productivity as compared to BBM. These cells expressed four folds higher lipid productivity (126 ± 5.54 mg/l/d) as compared to BBM. Cells cultivated in recycled DABE showed large sized lipid droplets accumulating 54.12 % of lipid content. Decrement in carbohydrate (17.76 %) and protein content (28.12 %) with loss of photosynthetic pigments compared to BBM grown cells were also recorded. The fatty acid profiles of cells cultivated in recycled DABE revealed the dominance of C16:0 (39.66 %), C18:1 (29.41 %) and C18:0 (15.82 %), respectively. This model is self-sustained and aims at neutralizing excessive feedstock consumption by exploiting recycled de-oiled algal biomass for cultivation of microalgae, making the process cost effective.

Algal Biomass Analysis by Laser-Based Analytical Techniques—A Review

PubMed Central

Pořízka, Pavel; Prochazková, Petra; Prochazka, David; Sládková, Lucia; Novotný, Jan; Petrilak, Michal; Brada, Michal; Samek, Ota; Pilát, Zdeněk; Zemánek, Pavel; Adam, Vojtěch; Kizek, René; Novotný, Karel; Kaiser, Jozef

2014-01-01

Algal biomass that is represented mainly by commercially grown algal strains has recently found many potential applications in various fields of interest. Its utilization has been found advantageous in the fields of bioremediation, biofuel production and the food industry. This paper reviews recent developments in the analysis of algal biomass with the main focus on the Laser-Induced Breakdown Spectroscopy, Raman spectroscopy, and partly Laser-Ablation Inductively Coupled Plasma techniques. The advantages of the selected laser-based analytical techniques are revealed and their fields of use are discussed in detail. PMID:25251409

A bio-anodic filter facilitated entrapment, decomposition and in situ oxidation of algal biomass in wastewater effluent.

PubMed

Mohammadi Khalfbadam, Hassan; Cheng, Ka Yu; Sarukkalige, Ranjan; Kaksonen, Anna H; Kayaalp, Ahmet S; Ginige, Maneesha P

2016-09-01

This study examined for the first time the use of bioelectrochemical systems (BES) to entrap, decompose and oxidise fresh algal biomass from an algae-laden effluent. The experimental process consisted of a photobioreactor for a continuous production of the algal-laden effluent, and a two-chamber BES equipped with anodic graphite granules and carbon-felt to physically remove and oxidise algal biomass from the influent. Results showed that the BES filter could retain ca. 90% of the suspended solids (SS) loaded. A coulombic efficiency (CE) of 36.6% (based on particulate chemical oxygen demand (PCOD) removed) was achieved, which was consistent with the highest CEs of BES studies (operated in microbial fuel cell mode (MFC)) that included additional pre-treatment steps for algae hydrolysis. Overall, this study suggests that a filter type BES anode can effectively entrap, decompose and in situ oxidise algae without the need for a separate pre-treatment step. Copyright © 2016 Elsevier Ltd. All rights reserved.

Swimming under the Influence: Effect of Algal Toxins on the Behavior of the Marine Ciliate Favella sp.

NASA Astrophysics Data System (ADS)

Sterling, A.; Echevarria, M. L.; Borrett, S. R.; Taylor, A. R.

2016-02-01

Although it is known that microzooplankton can regulate harmful algal bloom (HAB) dynamics through grazing of algae, the effects of HAB-related toxins on these micrograzers are unknown. Therefore I examined the effects of the algal toxins domoic acid (DA), brevetoxin (PbTx-2), and 2,4-trans,trans-decadienal (DDA) on the swimming behavior of the marine ciliate Favella sp. Neither DA nor PbTx-2 had a significant effect at the highest concentrations tested (800 nM and 400 nM respectively). However, about 50% of ciliates ceased swimming after 1 h exposure to 30 µM and 50 µM DDA and displayed significant behavioral changes within 5 min. Preliminary recovery experiments showed that up to 80% of the non-swimming ciliates were viable after 24 h, suggesting in these ciliates DDA did not induce programmed cell death. This work demonstrates that some, but not all, algal toxins may compromise the ability of microzooplankton to evade predators, capture prey, and regulate HABs.

Microfluidic engineered high cell density three-dimensional neural cultures

NASA Astrophysics Data System (ADS)

Cullen, D. Kacy; Vukasinovic, Jelena; Glezer, Ari; La Placa, Michelle C.

2007-06-01

Three-dimensional (3D) neural cultures with cells distributed throughout a thick, bioactive protein scaffold may better represent neurobiological phenomena than planar correlates lacking matrix support. Neural cells in vivo interact within a complex, multicellular environment with tightly coupled 3D cell-cell/cell-matrix interactions; however, thick 3D neural cultures at cell densities approaching that of brain rapidly decay, presumably due to diffusion limited interstitial mass transport. To address this issue, we have developed a novel perfusion platform that utilizes forced intercellular convection to enhance mass transport. First, we demonstrated that in thick (>500 µm) 3D neural cultures supported by passive diffusion, cell densities <=5.0 × 103 cells mm-3 were required for survival. In 3D neuronal and neuronal-astrocytic co-cultures with increased cell density (>=104 cells mm-3), continuous medium perfusion at 2.0-11.0 µL min-1 improved viability compared to non-perfused cultures (p < 0.01), which exhibited widespread cell death and matrix degradation. In perfused cultures, survival was dependent on proximity to the perfusion source at 2.00-6.25 µL min-1 (p < 0.05); however, at perfusion rates of 10.0-11.0 µL min-1 survival did not depend on the distance from the perfusion source, and resulted in a preservation of cell density with >90% viability in both neuronal cultures and neuronal-astrocytic co-cultures. This work demonstrates the utility of forced interstitial convection in improving the survival of high cell density 3D engineered neural constructs and may aid in the development of novel tissue-engineered systems reconstituting 3D cell-cell/cell-matrix interactions.

A novel closed cell culture device for fabrication of corneal epithelial cell sheets.

PubMed

Nakajima, Ryota; Kobayashi, Toyoshige; Moriya, Noboru; Mizutani, Manabu; Kan, Kazutoshi; Nozaki, Takayuki; Saitoh, Kazuo; Yamato, Masayuki; Okano, Teruo; Takeda, Shizu

2015-11-01

Automation technology for cell sheet-based tissue engineering would need to optimize the cell sheet fabrication process, stabilize cell sheet quality and reduce biological contamination risks. Biological contamination must be avoided in clinical settings. A closed culture system provides a solution for this. In the present study, we developed a closed culture device called a cell cartridge, to be used in a closed cell culture system for fabricating corneal epithelial cell sheets. Rabbit limbal epithelial cells were cultured on the surface of a porous membrane with 3T3 feeder cells, which are separate from the epithelial cells in the cell cartridges and in the cell-culture inserts as a control. To fabricate the stratified cell sheets, five different thicknesses of the membranes which were welded to the cell cartridge, were examined. Multilayered corneal epithelial cell sheets were fabricated in cell cartridges that were welded to a 25 µm-thick gas-permeable membrane, which was similar to the results with the cell-culture inserts. However, stratification of corneal epithelial cell sheets did not occur with cell cartridges that were welded to 100-300 µm-thick gas-permeable membranes. The fabricated cell sheets were evaluated by histological analyses to examine the expression of corneal epithelial-specific markers. Immunohistochemical analyses showed that a putative stem cell marker, p63, a corneal epithelial differentiation maker, CK3, and a barrier function marker, Claudin-1, were expressed in the appropriate position in the cell sheets. These results suggest that the cell cartridge is effective for fabricating corneal epithelial cell sheets. Copyright © 2012 John Wiley & Sons, Ltd.

Biona-C Cell Culture pH Monitoring System

NASA Technical Reports Server (NTRS)

Friedericks, C.

1999-01-01

Sensors 2000! is developing a system to demonstrate the ability to perform accurate, real-time measurements of pH and CO2 in a cell culture media in Space. The BIONA-C Cell Culture pH Monitoring System consists of S2K! developed ion selective sensors and control electronics integrated with the fluidics of a cell culture system. The integrated system comprises a "rail" in the Cell Culture Module (CCM) of WRAIR (Space Biosciences of Walter Read Army Institute of Research). The CCM is a Space Shuttle mid-deck locker experiment payload. The BIONA-C is displayed along with associated graphics and text explanations. The presentation will stimulate interest in development of sensor technology for real-time cell culture measurements. The transfer of this technology to other applications will also be of interest. Additional information is contained in the original document.

Retinoic acid stability in stem cell cultures.

PubMed

Sharow, Kyle A; Temkin, Boris; Asson-Batres, Mary Ann

2012-01-01

It has been reported that retinoids, such as retinoic acid (RA) and retinol (ROL), dissolved in aqueous solutions are susceptible to oxidative damage when exposed to light, air, and relatively high temperatures, conditions that are normal for culturing stem cells. Thus, questions arise regarding the interpretation of results obtained from studies of mouse embryonic stem cells exposed to retinoids because their isomerization state, their stability in culture conditions, and their interactions with other potential differentiation factors in growth media could influence developmental processes under study. Media samples were supplemented with retinoids and exposed to cell culture conditions with and without mouse embryonic stem cells (mESC), and retinoids were extracted and analyzed using HPLC. To determine whether retinoids are stable in media supplemented with fetal bovine serum (FBS) or in chemically-defined, serum-free media, mESC adapted to each type of growth media were investigated. Studies reported here indicate there was little loss or isomerization of at-RA, 9-cis-RA, 13-cis-RA, or ROL in cell cultures grown in serum-supplemented media when cell cultures were maintained in the dark and manipulated and observed under yellow light. In contrast, the stability of both at-RA and ROL were determined to be greatly reduced in serum-free media as compared with serum-supplemented media. Addition of 6 mg/ml bovine serum albumin was found to stabilize retinoids in serum-free media. It was also determined that ROL is less stable than RA in cell culture conditions.

Rotary orbital suspension culture of embryonic stem cell-derived neural stem/progenitor cells: impact of hydrodynamic culture on aggregate yield, morphology and cell phenotype.

PubMed

Laundos, Tiago L; Silva, Joana; Assunção, Marisa; Quelhas, Pedro; Monteiro, Cátia; Oliveira, Carla; Oliveira, Maria J; Pêgo, Ana P; Amaral, Isabel F

2017-08-01

Embryonic stem (ES)-derived neural stem/progenitor cells (ES-NSPCs) constitute a promising cell source for application in cell therapies for the treatment of central nervous system disorders. In this study, a rotary orbital hydrodynamic culture system was applied to single-cell suspensions of ES-NSPCs, to obtain homogeneously-sized ES-NSPC cellular aggregates (neurospheres). Hydrodynamic culture allowed the formation of ES-NSPC neurospheres with a narrower size distribution than statically cultured neurospheres, increasing orbital speeds leading to smaller-sized neurospheres and higher neurosphere yield. Neurospheres formed under hydrodynamic conditions (72 h at 55 rpm) showed higher cell compaction and comparable percentages of viable, dead, apoptotic and proliferative cells. Further characterization of cellular aggregates provided new insights into the effect of hydrodynamic shear on ES-NSPC behaviour. Rotary neurospheres exhibited reduced protein levels of N-cadherin and β-catenin, and higher deposition of laminin (without impacting fibronectin deposition), matrix metalloproteinase-2 (MMP-2) activity and percentage of neuronal cells. In line with the increased MMP-2 activity levels found, hydrodynamically-cultured neurospheres showed higher outward migration on laminin. Moreover, when cultured in a 3D fibrin hydrogel, rotary neurospheres generated an increased percentage of neuronal cells. In conclusion, the application of a constant orbital speed to single-cell suspensions of ES-NSPCs, besides allowing the formation of homogeneously-sized neurospheres, promoted ES-NSPC differentiation and outward migration, possibly by influencing the expression of cell-cell adhesion molecules and the secretion of proteases/extracellular matrix proteins. These findings are important when establishing the culture conditions needed to obtain uniformly-sized ES-NSPC aggregates, either for use in regenerative therapies or in in vitro platforms for biomaterial development or

INCIDENCE AND DETECTION OF PLEUROPNEUMONIA-LIKE ORGANISMS IN CELL CULTURES BY FLUORESCENT ANTIBODY AND CULTURAL PROCEDURES1

PubMed Central

Barile, Michael F.; Malizia, Walter F.; Riggs, Donald B.

1962-01-01

Barile, Michael F. (National Institutes of Health, Bethesda, Md.), Walter F. Malizia, and Donald B. Riggs. Incidence and detection of pleuropneumonia-like organisms in cell cultures by fluorescent antibody and cultural procedures. J. Bacteriol. 84:130–136. 1962—A total of 102 tissue-cell cultures from 17 separate laboratories was examined for pleuropneumonia-like organisms (PPLO) by the fluorescent antibody and cultural procedures. PPLO were isolated from 48 of the 49 tissue-cell cultures found positive for PPLO by the fluorescent antibody procedure, and results of the two procedures agreed in 101 of the 102 (99%) cases. PPLO were isolated from none of 10 primary-cell cultures prepared from six animal species and from 48 of 92 (52%) continuous-cell cultures prepared from eight animal species. Cells grown in media containing antibiotics were more frequently contaminated with PPLO (72%) than cells grown in antibiotic-free media (7%). Cultures (91%) from tissue-culture-producing laboratories and cultures (76%) used for propagation of microorganisms were contaminated with PPLO, although none used for tissue-culture metabolic studies was contaminated. In addition, our findings support the view that PPLO contamination of cell cultures is probably owing to bacterial contaminants which revert to L forms in the presence of antibiotics. Images PMID:13865001

Cell sources for in vitro human liver cell culture models.

PubMed

Zeilinger, Katrin; Freyer, Nora; Damm, Georg; Seehofer, Daniel; Knöspel, Fanny

2016-09-01

In vitro liver cell culture models are gaining increasing importance in pharmacological and toxicological research. The source of cells used is critical for the relevance and the predictive value of such models. Primary human hepatocytes (PHH) are currently considered to be the gold standard for hepatic in vitro culture models, since they directly reflect the specific metabolism and functionality of the human liver; however, the scarcity and difficult logistics of PHH have driven researchers to explore alternative cell sources, including liver cell lines and pluripotent stem cells. Liver cell lines generated from hepatomas or by genetic manipulation are widely used due to their good availability, but they are generally altered in certain metabolic functions. For the past few years, adult and pluripotent stem cells have been attracting increasing attention, due their ability to proliferate and to differentiate into hepatocyte-like cells in vitro However, controlling the differentiation of these cells is still a challenge. This review gives an overview of the major human cell sources under investigation for in vitro liver cell culture models, including primary human liver cells, liver cell lines, and stem cells. The promises and challenges of different cell types are discussed with a focus on the complex 2D and 3D culture approaches under investigation for improving liver cell functionality in vitro Finally, the specific application options of individual cell sources in pharmacological research or disease modeling are described. © 2016 by the Society for Experimental Biology and Medicine.

Density gradient electrophoresis of cultured human embryonic kidney cells

NASA Technical Reports Server (NTRS)

Plank, L. D.; Kunze, M. E.; Giranda, V.; Todd, P. W.

1985-01-01

Ground based confirmation of the electrophoretic heterogeneity of human embryonic kidney cell cultures, the general characterization of their electrophoretic migration, and observations on the general properties of cultures derived from electrophoretic subpopulations were studied. Cell migration in a density gradient electrophoresis column and cell electrophoretic mobility was determined. The mobility and heterogeneity of cultured human embryonic kidney cells with those of fixed rat erythrocytes as model test particle was compared. Electrophoretically separated cell subpopulations with respect to size, viability, and culture characteristics were examined.

Characterization of Intracellular and Extracellular Saxitoxin Levels in Both Field and Cultured Alexandrium spp. Samples from Sequim Bay, Washington

PubMed Central

Lefebvre, Kathi A.; Bill, Brian D.; Erickson, Aleta; Baugh, Keri A.; O’Rourke, Lohna; Costa, Pedro R.; Nance, Shelly; Trainer, Vera L.

2008-01-01

Traditionally, harmful algal bloom studies have primarily focused on quantifying toxin levels contained within the phytoplankton cells of interest. In the case of paralytic shellfish poisoning toxins (PSTs), intracellular toxin levels and the effects of dietary consumption of toxic cells by planktivores have been well documented. However, little information is available regarding the levels of extracellular PSTs that may leak or be released into seawater from toxic cells during blooms. In order to fully evaluate the risks of harmful algal bloom toxins in the marine food web, it is necessary to understand all potential routes of exposure. In the present study, extracellular and intracellular PST levels were measured in field seawater samples (collected weekly from June to October 2004–2007) and in Alexandrium spp. culture samples isolated from Sequim Bay, Washington. Measurable levels of intra- and extra-cellular toxins were detected in both field and culture samples via receptor binding assay (RBA) and an enzyme-linked immunosorbent assay (ELISA). Characterization of the PST toxin profile in the Sequim Bay isolates by pre-column oxidation and HPLC-fluorescence detection revealed that gonyautoxin 1 and 4 made up 65 ± 9.7 % of the total PSTs present. Collectively, these data confirm that extracellular PSTs are present during blooms of Alexandrium spp. in the Sequim Bay region. PMID:18728762

Achieving pH control in microalgal cultures through fed-batch addition of stoichiometrically-balanced growth media

PubMed Central

2013-01-01

Background Lack of accounting for proton uptake and secretion has confounded interpretation of the stoichiometry of photosynthetic growth of algae. This is also problematic for achieving growth of microalgae to high cell concentrations which is necessary to improve productivity and the economic feasibility of commercial-scale chemical production systems. Since microalgae are capable of consuming both nitrate and ammonium, this represents an opportunity to balance culture pH based on a nitrogen feeding strategy that does not utilize gas-phase CO2 buffering. Stoichiometry suggests that approximately 36 weight%N-NH4+ (balance nitrogen as NO3-) would minimize the proton imbalance and permit high-density photoautotrophic growth as it does in higher plant tissue culture. However, algal media almost exclusively utilize nitrate, and ammonium is often viewed as ‘toxic’ to algae. Results The microalgae Chlorella vulgaris and Chlamydomonas reinhardtii exclusively utilize ammonium when both ammonium and nitrate are provided during growth on excess CO2. The resulting proton imbalance from preferential ammonium utilization causes the pH to drop too low to sustain further growth when ammonium was only 9% of the total nitrogen (0.027 gN-NH4+/L). However, providing smaller amounts of ammonium sequentially in the presence of nitrate maintained the pH of a Chlorella vulgaris culture for improved growth on 0.3 gN/L to 5 gDW/L under 5% CO2 gas-phase supplementation. Bioreactor pH dynamics are shown to be predictable based on simple nitrogen assimilation as long as there is sufficient CO2 availability. Conclusions This work provides both a media formulation and a feeding strategy with a focus on nitrogen metabolism and regulation to support high-density algal culture without buffering. The instability in culture pH that is observed in microalgal cultures in the absence of buffers can be overcome through alternating utilization of ammonium and nitrate. Despite the highly regulated

21 CFR 864.2280 - Cultured animal and human cells.

Code of Federal Regulations, 2012 CFR

2012-04-01

... 21 Food and Drugs 8 2012-04-01 2012-04-01 false Cultured animal and human cells. 864.2280 Section... Cultured animal and human cells. (a) Identification. Cultured animal and human cells are in vitro cultivated cell lines from the tissue of humans or other animals which are used in various diagnostic...

21 CFR 864.2280 - Cultured animal and human cells.

Code of Federal Regulations, 2013 CFR

2013-04-01

... 21 Food and Drugs 8 2013-04-01 2013-04-01 false Cultured animal and human cells. 864.2280 Section... Cultured animal and human cells. (a) Identification. Cultured animal and human cells are in vitro cultivated cell lines from the tissue of humans or other animals which are used in various diagnostic...

21 CFR 864.2280 - Cultured animal and human cells.

Code of Federal Regulations, 2014 CFR

2014-04-01

... 21 Food and Drugs 8 2014-04-01 2014-04-01 false Cultured animal and human cells. 864.2280 Section... Cultured animal and human cells. (a) Identification. Cultured animal and human cells are in vitro cultivated cell lines from the tissue of humans or other animals which are used in various diagnostic...

21 CFR 864.2280 - Cultured animal and human cells.

Code of Federal Regulations, 2011 CFR

2011-04-01

... 21 Food and Drugs 8 2011-04-01 2011-04-01 false Cultured animal and human cells. 864.2280 Section... Cultured animal and human cells. (a) Identification. Cultured animal and human cells are in vitro cultivated cell lines from the tissue of humans or other animals which are used in various diagnostic...

21 CFR 876.5885 - Tissue culture media for human ex vivo tissue and cell culture processing applications.

Code of Federal Regulations, 2010 CFR

2010-04-01

... cell culture processing applications. 876.5885 Section 876.5885 Food and Drugs FOOD AND DRUG... DEVICES Therapeutic Devices § 876.5885 Tissue culture media for human ex vivo tissue and cell culture processing applications. (a) Identification. Tissue culture media for human ex vivo tissue and cell culture...

Copper desorption from Gelidium algal biomass.

PubMed

Vilar, Vítor J P; Botelho, Cidália M S; Boaventura, Rui A R

2007-04-01

Desorption of divalent copper from marine algae Gelidium sesquipedale, an algal waste (from agar extraction industry) and a composite material (the algal waste immobilized in polyacrylonitrile) was studied in a batch system. Copper ions were first adsorbed until saturation and then desorbed by HNO(3) and Na(2)EDTA solutions. Elution efficiency using HNO(3) increases as pH decreases. At pH=1, for a solid to liquid ratio S/L=4gl(-1), elution efficiency was 97%, 95% and 88%, the stoichiometric coefficient for the ionic exchange, 0.70+/-0.02, 0.73+/-0.05 and 0.76+/-0.06 and the selectivity coefficient, 0.93+/-0.07, 1.0+/-0.3 and 1.1+/-0.3, respectively, for algae Gelidium, algal waste and composite material. Complexation of copper ions by EDTA occurs in a molar proportion of 1:1 and the elution efficiency increases with EDTA concentration. For concentrations of 1.4, 0.88 and 0.57 mmoll(-1), the elution efficiency for S/L=4gl(-1), was 91%, 86% and 78%, respectively, for algae Gelidium, algal waste and composite material. The S/L ratio, in the range 1-20gl(-1), has little influence on copper recovery by using 0.1M HNO(3). Desorption kinetics was very fast for all biosorbents. Kinetic data using HNO(3) as eluant were well described by the mass transfer model, considering the average metal concentration in the solid phase and the equilibrium relationship given by the mass action law. The homogeneous diffusion coefficient varied between 1.0 x 10(-7)cm(2)s(-1) for algae Gelidium and 3.0 x 10(-7)cm(2)s(-1) for the composite material.

Three dimensional spheroid cell culture for nanoparticle safety testing.

PubMed

Sambale, Franziska; Lavrentieva, Antonina; Stahl, Frank; Blume, Cornelia; Stiesch, Meike; Kasper, Cornelia; Bahnemann, Detlef; Scheper, Thomas

2015-07-10

Nanoparticles are widely employed for many applications and the number of consumer products, incorporating nanotechnology, is constantly increasing. A novel area of nanotechnology is the application in medical implants. The widespread use of nanoparticles leads to their higher prevalence in our environment. This, in turn, raises concerns regarding potential risks to humans. Previous studies have shown possible hazardous effects of some nanoparticles on mammalian cells grown in two-dimensional (2D) cultures. However, 2D in vitro cell cultures display several disadvantages such as changes in cell shape, cell function, cell responses and lack of cell-cell contacts. For this reason, the development of better models for mimicking in vivo conditions is essential. In the present work, we cultivated A549 cells and NIH-3T3 cells in three-dimensional (3D) spheroids and investigated the effects of zinc oxide (ZnO-NP) and titanium dioxide nanoparticles (TiO2-NP). The results were compared to cultivation in 2D monolayer culture. A549 cells in 3D cell culture formed loose aggregates which were more sensitive to the toxicity of ZnO-NP in comparison to cells grown in 2D monolayers. In contrast, NIH-3T3 cells showed a compact 3D spheroid structure and no differences in the sensitivity of the NIH-3T3 cells to ZnO-NP were observed between 2D and 3D cultures. TiO2-NP were non-toxic in 2D cultures but affected cell-cell interaction during 3D spheroid formation of A549 and NIH-3T3 cells. When TiO2-NP were directly added during spheroid formation in the cultures of the two cell lines tested, several smaller spheroids were formed instead of a single spheroid. This effect was not observed if the nanoparticles were added after spheroid formation. In this case, a slight decrease in cell viability was determined only for A549 3D spheroids. The obtained results demonstrate the importance of 3D cell culture studies for nanoparticle safety testing, since some effects cannot be revealed in 2D

Culture of human mesenchymal stem cells using a candidate pharmaceutical grade xeno-free cell culture supplement derived from industrial human plasma pools.

PubMed

Díez, José M; Bauman, Ewa; Gajardo, Rodrigo; Jorquera, Juan I

2015-03-13

Fetal bovine serum (FBS) is an animal product used as a medium supplement. The animal origin of FBS is a concern if cultured stem cells are to be utilized for human cell therapy. Therefore, a substitute for FBS is desirable. In this study, an industrial, xeno-free, pharmaceutical-grade supplement for cell culture (SCC) under development at Grifols was tested for growth of human mesenchymal stem cells (hMSCs), cell characterization, and differentiation capacity. SCC is a freeze-dried product obtained through cold-ethanol fractionation of industrial human plasma pools from healthy donors. Bone marrow-derived hMSC cell lines were obtained from two commercial suppliers. Cell growth was evaluated by culturing hMSCs with commercial media or media supplemented with SCC or FBS. Cell viability and cell yield were assessed with an automated cell counter. Cell surface markers were studied by indirect immunofluorescence assay. Cells were cultured then differentiated into adipocytes, chondrocytes, osteoblasts, and neurons, as assessed by specific staining and microscopy observation. SCC supported the growth of commercial hMSCs. Starting from the same number of seeded cells in two consecutive passages of culture with medium supplemented with SCC, hMSC yield and cell population doubling time were equivalent to the values obtained with the commercial medium and was consistent among lots. The viability of hMSCs was higher than 90%, while maintaining the characteristic phenotype of undifferentiated hMSCs (positive for CD29, CD44, CD90, CD105, CD146, CD166 and Stro-1; negative for CD14 and CD19). Cultured hMSCs maintained the potential for differentiation into adipocytes, chondrocytes, osteoblasts, and neurons. The tested human plasma-derived SCC sustains the adequate growth of hMSCs, while preserving their differentiation capacity. SCC can be a potential candidate for cell culture supplement in advanced cell therapies.

Two-decade reconstruction of algal blooms in China's Lake Taihu.

PubMed

Duan, Hongtao; Ma, Ronghua; Xu, Xiaofeng; Kong, Fanxiang; Zhang, Shouxuan; Kong, Weijuan; Hao, Jingyan; Shang, Linlin

2009-05-15

The algal blooming in the inland lakes has become a critically important issue for its impacts not only on local natural and social environments, but also on global human community. However, the occurrences of blooming on larger spatial scale and longer time scale have rarely been studied. As the third largest freshwater lake in China, Lake Taihu has drawn increasing attention from both public and scientific communities concerning its degradation. Using available satellite images, we reconstructed the spatial and temporal patterns of algal blooms in Lake Taihu through the pasttwo decades. The blooming characteristics over the past two decades were examined with the dynamic of initial blooming date being highlighted. The initial blooming dates were gradually becoming later and later from 1987 to 1997. Since 1998, however, the initial blooming date came earlier and earlier year by year, with approximately 11.42 days advancement per year. From 1987 to 2007, the annual duration of algal blooms lengthened year by year, in line with the substantial increases in the occurrences of algal blooms in spring and summer months. The algal blooms usually occur in northern bays and spread to center and south parts of Lake Taihu. The increases in previous winter's mean daily minimum temperature partially contributed to the earlier blooming onset. However, human activities, expressed as total gross domestic product (GDP) and population, outweighed the climatic contribution on the initial blooming date and blooming duration. This study may provide insights for the policy makers who try to curb the algal blooming and improve the water quality of inland freshwater lakes.

Differential marker expression by cultures rich in mesenchymal stem cells

PubMed Central

2013-01-01

Background Mesenchymal stem cells have properties that make them amenable to therapeutic use. However, the acceptance of mesenchymal stem cells in clinical practice requires standardized techniques for their specific isolation. To date, there are no conclusive marker (s) for the exclusive isolation of mesenchymal stem cells. Our aim was to identify markers differentially expressed between mesenchymal stem cell and non-stem cell mesenchymal cell cultures. We compared and contrasted the phenotype of tissue cultures in which mesenchymal stem cells are rich and rare. By initially assessing mesenchymal stem cell differentiation, we established that bone marrow and breast adipose cultures are rich in mesenchymal stem cells while, in our hands, foreskin fibroblast and olfactory tissue cultures contain rare mesenchymal stem cells. In particular, olfactory tissue cells represent non-stem cell mesenchymal cells. Subsequently, the phenotype of the tissue cultures were thoroughly assessed using immuno-fluorescence, flow-cytometry, proteomics, antibody arrays and qPCR. Results Our analysis revealed that all tissue cultures, regardless of differentiation potential, demonstrated remarkably similar phenotypes. Importantly, it was also observed that common mesenchymal stem cell markers, and fibroblast-associated markers, do not discriminate between mesenchymal stem cell and non-stem cell mesenchymal cell cultures. Examination and comparison of the phenotypes of mesenchymal stem cell and non-stem cell mesenchymal cell cultures revealed three differentially expressed markers – CD24, CD108 and CD40. Conclusion We indicate the importance of establishing differential marker expression between mesenchymal stem cells and non-stem cell mesenchymal cells in order to determine stem cell specific markers. PMID:24304471

Chromosomal instability and telomere shortening in long-term culture of hematopoietic stem cells: insights from a cell culture model of RPS14 haploinsufficiency.

PubMed

Thomay, K; Schienke, A; Vajen, B; Modlich, U; Schambach, A; Hofmann, W; Schlegelberger, B; Göhring, G

2014-01-01

The fate of cultivated primary hematopoietic stem cells (HSCs) with respect to genetic instability and telomere attrition has not yet been described in great detail. Thus, knowledge of the genetic constitution of HSCs is important when interpreting results of HSCs in culture. While establishing a cell culture model for myelodysplastic syndrome with a deletion in 5q by performing RPS14 knockdown, we found surprising data that may be of importance for any CD34+ cell culture experiments. We performed cytogenetic analyses and telomere length measurement on transduced CD34+ cells and untransduced control cells to observe the effects of long-term culturing. Initially, CD34+ cells had a normal median telomere length of about 12 kb and showed no signs of chromosomal instability. During follow-up, the median telomere length seemed to decrease and, simultaneously, increased chromosomal instability could be observed - in modified and control cells. One culture showed a clonal monosomy 7 - independent of prior RPS14 knockdown. During further culturing, it seemed that the telomeres re-elongated, and chromosomes stabilized, while TERT expression was not elevated. In summary, irrespective of our results of RPS14 knockdown in the long-term culture of CD34+ cells, it becomes clear that cell culture artefacts inducing telomere shortening and chromosomal instability have to be taken into account and regular cytogenetic analyses should always be performed. © 2013 S. Karger AG, Basel.

Introducing Mammalian Cell Culture and Cell Viability Techniques in the Undergraduate Biology Laboratory †

PubMed Central

Bowey-Dellinger, Kristen; Dixon, Luke; Ackerman, Kristin; Vigueira, Cynthia; Suh, Yewseok K.; Lyda, Todd; Sapp, Kelli; Grider, Michael; Crater, Dinene; Russell, Travis; Elias, Michael; Coffield, V. McNeil; Segarra, Verónica A.

2017-01-01

Undergraduate students learn about mammalian cell culture applications in introductory biology courses. However, laboratory modules are rarely designed to provide hands-on experience with mammalian cells or teach cell culture techniques, such as trypsinization and cell counting. Students are more likely to learn about cell culture using bacteria or yeast, as they are typically easier to grow, culture, and manipulate given the equipment, tools, and environment of most undergraduate biology laboratories. In contrast, the utilization of mammalian cells requires a dedicated biological safety cabinet and rigorous antiseptic techniques. For this reason, we have devised a laboratory module and method herein that familiarizes students with common cell culture procedures, without the use of a sterile hood or large cell culture facility. Students design and perform a time-efficient inquiry-based cell viability experiment using HeLa cells and tools that are readily available in an undergraduate biology laboratory. Students will become familiar with common techniques such as trypsinizing cells, cell counting with a hemocytometer, performing serial dilutions, and determining cell viability using trypan blue dye. Additionally, students will work with graphing software to analyze their data and think critically about the mechanism of death on a cellular level. Two different adaptations of this inquiry-based lab are presented—one for non-biology majors and one for biology majors. Overall, these laboratories aim to expose students to mammalian cell culture and basic techniques and help them to conceptualize their application in scientific research. PMID:28861134

Foetal hepatic progenitor cells assume a cholangiocytic cell phenotype during two-dimensional pre-culture.

PubMed

Anzai, Kazuya; Chikada, Hiromi; Tsuruya, Kota; Ida, Kinuyo; Kagawa, Tatehiro; Inagaki, Yutaka; Mine, Tesuya; Kamiya, Akihide

2016-06-23

Liver consists of parenchymal hepatocytes and other cells. Liver progenitor cell (LPC) is the origin of both hepatocytes and cholangiocytic cells. The analyses of mechanism regulating differentiation of LPCs into these functional cells are important for liver regenerative therapy using progenitor cells. LPCs in adult livers were found to form cysts with cholangiocytic characteristics in 3D culture. In contrast, foetal LPCs cannot form these cholangiocytic cysts in the same culture. Thus, the transition of foetal LPCs into cholangiocytic progenitor cells might occur during liver development. Primary CD45(-)Ter119(-)Dlk1(+) LPCs derived from murine foetal livers formed ALBUMIN (ALB)(+)CYTOKERATIN (CK)19(-) non-cholangiocytic cysts within 3D culture. In contrast, when foetal LPCs were pre-cultured on gelatine-coated dishes, they formed ALB(-)CK19(+) cholangiocytic cysts. When hepatocyte growth factor or oncostatin M, which are inducers of hepatocytic differentiation, was added to pre-culture, LPCs did not form cholangiocytic cysts. These results suggest that the pre-culture on gelatine-coated dishes changed the characteristics of foetal LPCs into cholangiocytic cells. Furthermore, neonatal liver progenitor cells were able to form cholangiocytic cysts in 3D culture without pre-culture. It is therefore possible that the pre-culture of mid-foetal LPCs in vitro functioned as a substitute for the late-foetal maturation step in vivo.

A single-cell and feeder-free culture system for monkey embryonic stem cells.

PubMed

Ono, Takashi; Suzuki, Yutaka; Kato, Yosuke; Fujita, Risako; Araki, Toshihiro; Yamashita, Tomoko; Kato, Hidemasa; Torii, Ryuzo; Sato, Naoya

2014-01-01

Primate pluripotent stem cells (PSCs), including embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs), hold great potential for research and application in regenerative medicine and drug discovery. To maximize primate PSC potential, a practical system is required for generating desired functional cells and reproducible differentiation techniques. Much progress regarding their culture systems has been reported to date; however, better methods would still be required for their practical use, particularly in industrial and clinical fields. Here we report a new single-cell and feeder-free culture system for primate PSCs, the key feature of which is an originally formulated serum-free medium containing FGF and activin. In this culture system, cynomolgus monkey ESCs can be passaged many times by single-cell dissociation with traditional trypsin treatment and can be propagated with a high proliferation rate as a monolayer without any feeder cells; further, typical PSC properties and genomic stability can be retained. In addition, it has been demonstrated that monkey ESCs maintained in the culture system can be used for various experiments such as in vitro differentiation and gene manipulation. Thus, compared with the conventional culture system, monkey ESCs grown in the aforementioned culture system can serve as a cell source with the following practical advantages: simple, stable, and easy cell maintenance; gene manipulation; cryopreservation; and desired differentiation. We propose that this culture system can serve as a reliable platform to prepare primate PSCs useful for future research and application.

Biogenic synthesis of silver nanoparticle by using secondary metabolites from Pseudomonas aeruginosa DM1 and its anti-algal effect on Chlorella vulgaris and Chlorella pyrenoidosa.

PubMed

Kumari, Rima; Barsainya, Manjari; Singh, Devendra Pratap

2017-02-01

Biogenic synthesis of silver nanoparticles (AgNPs) using extracellular metabolites from the bacterium Pseudomonas aeruginosa DM1 offers an eco-friendly and sustainable way of metal nanoparticle synthesis. The present work highlights the biotransformation of silver nitrate solution into AgNP, mediated by extracellular secondary metabolite pyoverdine, a siderophore produced by P. aeruginosa. The bioreduction of silver ions into AgNPs by using pyoverdine was recorded in terms of Fourier transform infrared spectroscopy (FTIR) analysis and color change in the reaction mixture (AgNO 3 + pyoverdine) from pale yellow to dark brown with absorption maxima at 415 nm. The results of X-ray diffraction (XRD) analysis of AgNPs showed its crystalline face-centered cubic structure. The scanning electron microscopy (SEM) and transmission electron microscopy (TEM) pictures of AgNPs showed spherical morphology of AgNP in the range of 45-100 nm, with tendency of agglomerations. The energy-dispersive X-ray (EDX) analysis of particles provided strong signal of elemental silver with few minor peaks of other impurities. The present approach offers a unique in vitro method of metal nanoparticle synthesis by exogenously produced bacterial secondary metabolites, where direct contact between the toxic metal and biological resource material can be avoided. The biologically synthesized AgNPs are found to have anti-algal effects against two species of Chlorella (Chlorella vulgaris and Chlorella pyenoidosa), as indicated by zone of growth inhibition on algal culture plates. Further results exhibit concentration-dependent progressive inhibition of chlorophyll content in the algal cells by AgNPs, confirming the algicidal effect of AgNPs.

Using live algae at the anode of a microbial fuel cell to generate electricity.

PubMed

Xu, Chang; Poon, Karen; Choi, Martin M F; Wang, Ruihua

2015-10-01

Live green microalgae Chlorella pyrenoidosa was introduced in the anode of a microbial fuel cell (MFC) to act as an electron donor. By controlling the oxygen content, light intensity, and algal cell density at the anode, microalgae would generate electricity without requiring externally added substrates. Two models of algal microbial fuel cells (MFCs) were constructed with graphite/carbon electrodes and no mediator. Model 1 algal MFC has live microalgae grown at the anode and potassium ferricyanide at the cathode, while model 2 algal MFC had live microalgae in both the anode and cathode in different growth conditions. Results indicated that a higher current produced in model 1 algal MFC was obtained at low light intensity of 2500 lx and algal cell density of 5 × 10(6) cells/ml, in which high algal density would limit the electricity generation, probably by increasing oxygen level and mass transfer problem. The maximum power density per unit anode volume obtained in model 1 algal MFC was relatively high at 6030 mW/m(2), while the maximum power density at 30.15 mW/m(2) was comparable with that of previous reported bacteria-driven MFC with graphite/carbon electrodes. A much smaller power density at 2.5 mW/m(2) was observed in model 2 algal MFC. Increasing the algal cell permeability by 4-nitroaniline would increase the open circuit voltage, while the mitochondrial acting and proton leak promoting agents resveratrol and 2,4-dinitrophenol would increase the electric current production in algal MFC.

Cell Culture in Microgravity: Opening the Door to Space Cell Biology

NASA Technical Reports Server (NTRS)

Pellis, Neal R.; Dawson, David L. (Technical Monitor)

1999-01-01

Adaptational response of human cell populations to microgravity is investigated using simulation, short-term Shuttle experiments, and long-term microgravity. Simulation consists of a clinostatically-rotated cell culture system. The system is a horizontally-rotated cylinder completely filled with culture medium. Low speed rotation results in continuous-fall of the cells through the fluid medium. In this setting, cells: 1) aggregate, 2) propagate in three dimensions, 3) synthesize matrix, 4) differentiate, and 5) form sinusoids that facilitate mass transfer. Space cell culture is conducted in flight bioreactors and in static incubators. Cells grown in microgravity are: bovine cartilage, promyelocytic leukemia, kidney proximal tubule cells, adrenal medulla, breast and colon cancer, and endothelium. Cells were cultured in space to test specific hypotheses. Cartilage cells were used to determine structural differences in cartilage grown in space compared to ground-based bioreactors. Results from a 130-day experiment on Mir revealed that cartilage grown in space was substantially more compressible due to insufficient glycosaminoglycan in the matrix. Interestingly, earth-grown cartilage conformed better to the dimensions of the scaffolding material, while the Mir specimens were spherical. The other cell populations are currently being analyzed for cell surface properties, gene expression, and differentiation. Results suggest that some cells spontaneously differentiate in microgravity. Additionally, vast changes in gene expression may occur in response to microgravity. In conclusion, the transition to microgravity may constitute a physical perturbation in cells resulting in unique gene expressions, the consequences of which may be useful in tissue engineering, disease modeling, and space cell biology.

Method for delivery of small molecules and proteins across the cell wall of algae using molecular transporters

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

Geihe, Erika; Trantow, Brian; Wender, Paul

The introduction of tools to study, control or expand the inner-workings of algae has been slow to develop. Provided are embodiments of a molecular method based on guanidinium-rich molecular transporters (GR-MoTrs) for bringing molecular cargos into algal cells. The methods of the disclosure have been shown to work in wild-type algae that have an intact cell wall. Developed using Chlamydomonas reinhardtii, this method is also successful with less studied algae, including Neochloris oleoabundans and Scenedesmus dimorphus, thus providing a new and versatile tool for algal research and modification. The method of delivering a cargo compound to an algal cell comprisesmore » contacting an algal cell with a guanidinium-rich delivery vehicle comprising a guanidinium-rich molecular transporter (GR-MoTr) linked to a cargo compound desired to be delivered to the algal cell, whereby the guanidinium-rich molecular transporter can traverse the algal cell wall, thereby delivering the cargo compound to the algal cell.« less

Enrichment of spinal cord cell cultures with motoneurons

PubMed Central

1978-01-01

Spinal cord cell cultures contain several types of neurons. Two methods are described for enriching such cultures with motoneurons (defined here simply as cholinergic cells that are capable of innervating muscle). In the first method, 7-day embryonic chick spinal cord neurons were separated according to size by 1 g velocity sedimentation. It is assumed that cholinergic motoneurons are among the largest cells present at this stage. The spinal cords were dissociated vigorously so that 95-98% of the cells in the initial suspension were isolated from one another. Cells in leading fractions (large cell fractions: LCFs) contain about seven times as much choline acetyltransferase (CAT) activity per unit cytoplasm as do cells in trailing fractions (small cell fractions: SCFs). Muscle cultures seeded with LCFs develop 10-70 times as much CAT as cultures seeded with SCFs and six times as much CAT as cultures seeded with control (unfractionated) spinal cord cells. More than 20% of the large neurons in LCF-muscle cultures innervate nearby myotubes. In the second method, neurons were gently dissociated from 4-day embryonic spinal cords and maintained in vitro. This approach is based on earlier observations that cholinergic neurons are among the first cells to withdraw form the mitotic cycle in the developing chick embryo (Hamburger, V. 1948. J. Comp. Neurol. 88:221- 283; and Levi-Montalcini, R. 1950. J. Morphol. 86:253-283). 4-Day spinal cord-muscle cultures develop three times as much CAT as do 7-day spinal cord-muscle plates, prepared in the same (gentle) manner. More than 50% of the relatively large 4-day neurons innervate nearby myotubes. Thus, both methods are useful first steps toward the complete isolation of motoneurons. Both methods should facilitate study of the development of cholinergic neurons and of nerve-muscle synapse formation. PMID:566275

How do culture media influence in vitro perivascular cell behavior?

PubMed

Huber, Birgit; Volz, Ann-Cathrin; Kluger, Petra Juliane

2015-12-01

Perivascular cells are multilineage cells located around the vessel wall and important for wall stabilization. In this study, we evaluated a stem cell media and a perivascular cell-specific media for the culture of primary perivascular cells regarding their cell morphology, doubling time, stem cell properties, and expression of cell type-specific markers. When the two cell culture media were compared to each other, perivascular cells cultured in the stem cell medium had a more elongated morphology and a faster doubling rate and cells cultured in the pericyte medium had a more typical morphology, with several filopodia, and a slower doubling rate. To evaluate stem cell properties, perivascular cells, CD146(-) cells, and mesenchymal stem cells (MSCs) were differentiated into the adipogenic, osteogenic, and chondrogenic lineages. It was seen that perivascular cells, as well as CD146(-) cells and MSCs, cultured in stem cell medium showed greater differentiation than cells cultured in pericyte-specific medium. The expression of pericyte-specific markers CD146, neural/glial antigen 2 (NG2), platelet-derived growth factor receptor-β (PDGFR-β), myosin, and α-smooth muscle actin (α-SMA) could be found in both pericyte cultures, as well as to varying amounts in CD146(-) cells, MSCs, and endothelial cells. The here presented work shows that perivascular cells can adapt to their in vitro environment and cell culture conditions influence cell functionality, such as doubling rate or differentiation behavior. Pericyte-specific markers were shown to be expressed also from cells other than perivascular cells. We can further conclude that CD146(+) perivascular cells are inhomogeneous cell population probably containing stem cell subpopulations, which are located perivascular around capillaries. © 2015 International Federation for Cell Biology.

Biolistic transformation of cotton embryogenic cell suspension cultures

USDA-ARS?s Scientific Manuscript database

Genetic transformation of cotton is highly dependent on the ability to regenerate fertile plants from transgenic cells through somatic embryogenesis. Induction of embryogenic cell cultures is genotype-dependant. However, once embryogenic cell cultures are available, they can be effectively used fo...

Cyanobacterial-algal cenoses in ordinary chernozems under the impact of different phytoameliorants

NASA Astrophysics Data System (ADS)

Dubovik, I. E.; Suyundukov, Ya. T.; Khasanova, R. F.; Shalygina, R. R.

2016-04-01

General ecological and taxonomic characteristics of cyanobacterial-algal cenoses in ordinary chernozems under different ameliorative plants (phytoameliorants) were studied in the Trans-Ural region of the Republic of Bashkortostan. A comparative analysis of the taxa of studied cenoses in the soils under leguminous herbs and grasses was performed. The phytoameliorative effect of different herbs and their relationships with cyanobacterial-algal cenoses were examined. Overall, 134 cyanoprokaryotic and algal species belonging to 70 genera, 36 families, 15 orders, and 9 classes were identified. Cyanobacterial-algal cenoses included the divisions of Chlorophyta, Cyanoprokaryota, Xanthophyta, Bacillariophyta, and Euglenophyta. Representatives of Ch-, X-, CF-, and P-forms were the leading ecobiomorphs in the studied cenoses.

Bacterial Associates Modify Growth Dynamics of the Dinoflagellate Gymnodinium catenatum

PubMed Central

Bolch, Christopher J. S.; Bejoy, Thaila A.; Green, David H.

2017-01-01

Marine phytoplankton cells grow in close association with a complex microbial associate community known to affect the growth, behavior, and physiology of the algal host. The relative scale and importance these effects compared to other major factors governing algal cell growth remain unclear. Using algal-bacteria co-culture models based on the toxic dinoflagellate Gymnodinium catenatum, we tested the hypothesis that associate bacteria exert an independent effect on host algal cell growth. Batch co-cultures of G. catenatum were grown under identical environmental conditions with simplified bacterial communities composed of one-, two-, or three-bacterial associates. Modification of the associate community membership and complexity induced up to four-fold changes in dinoflagellate growth rate, equivalent to the effect of a 5°C change in temperature or an almost six-fold change in light intensity (20–115 moles photons PAR m-2 s-1). Almost three-fold changes in both stationary phase cell concentration and death rate were also observed. Co-culture with Roseobacter sp. DG874 reduced dinoflagellate exponential growth rate and led to a more rapid death rate compared with mixed associate community controls or co-culture with either Marinobacter sp. DG879, Alcanivorax sp. DG881. In contrast, associate bacteria concentration was positively correlated with dinoflagellate cell concentration during the exponential growth phase, indicating growth was limited by supply of dinoflagellate-derived carbon. Bacterial growth increased rapidly at the onset of declining and stationary phases due to either increasing availability of algal-derived carbon induced by nutrient stress and autolysis, or at mid-log phase in Roseobacter co-cultures potentially due to the onset of bacterial-mediated cell lysis. Co-cultures with the three bacterial associates resulted in dinoflagellate and bacterial growth dynamics very similar to more complex mixed bacterial community controls, suggesting that

The Regulation of Gene Expression in Cnidarian-Algal Associations.

DTIC Science & Technology

1998-07-13

symbiotic cnidarians , Aiptasia pallida, Anthopleura eligantissima, synbiosis-specific proteins, cDNA libraries, O. SECURITY CLASSIFICATION OP REPORT...gene expression in cnidarian -algal associations Award Period: 1 July 1995-30 June 1998 Objectives: A. To identify and characterize heat shock...Exploring Symbiosis-Specific Gene Expression in Cnidarian /Algal Associations. In: Molecular Approaches to the Study of the Ocean.. Ed. K. Cooksey, Chapman

Biogelx: Cell Culture on Self-Assembling Peptide Gels.

PubMed

Harper, Mhairi M; Connolly, Michael L; Goldie, Laura; Irvine, Eleanore J; Shaw, Joshua E; Jayawarna, Vineetha; Richardson, Stephen M; Dalby, Matthew J; Lightbody, David; Ulijn, Rein V

2018-01-01

Aromatic peptide amphiphiles can form self-supporting nanostructured hydrogels with tunable mechanical properties and chemical compositions. These hydrogels are increasingly applied in two-dimensional (2D) and three-dimensional (3D) cell culture, where there is a rapidly growing need to store, grow, proliferate, and manipulate naturally derived cells within a hydrated, 3D matrix. Biogelx Limited is a biomaterials company, created to commercialize these bio-inspired hydrogels to cell biologists for a range of cell culture applications. This chapter describes methods of various characterization and cell culture techniques specifically optimized for compatibility with Biogelx products.

2016 National Algal Biofuels Technology Review Fact Sheet

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

None

2016-06-01

Algae-based biofuels and bioproducts offer great promise in contributing to the U.S. Department of Energy (DOE) Bioenergy Technologies Office’s (BETO’s) vision of a thriving and sustainable bioeconomy fueled by innovative technologies. The state of technology for producing algal biofuels continues to mature with ongoing investment by DOE and the private sector, but additional research, development, and demonstration (RD&D) is needed to achieve widespread deployment of affordable, scalable, and sustainable algal biofuels.

Recombinant protein production and insect cell culture and process

NASA Technical Reports Server (NTRS)

Spaulding, Glenn (Inventor); Prewett, Tacey (Inventor); Goodwin, Thomas (Inventor); Francis, Karen (Inventor); Andrews, Angela (Inventor); Oconnor, Kim (Inventor)

1993-01-01

A process has been developed for recombinant production of selected polypeptides using transformed insect cells cultured in a horizontally rotating culture vessel modulated to create low shear conditions. A metabolically transformed insect cell line is produced using the culture procedure regardless of genetic transformation. The recombinant polypeptide can be produced by an alternative process using the cultured insect cells as host for a virus encoding the described polypeptide such as baculovirus. The insect cells can also be a host for viral production.

Effects of fertilizers used in agricultural fields on algal blooms

NASA Astrophysics Data System (ADS)

Chakraborty, Subhendu; Tiwari, P. K.; Sasmal, S. K.; Misra, A. K.; Chattopadhyay, Joydev

2017-06-01

The increasing occurrence of algal blooms and their negative ecological impacts have led to intensified monitoring activities. This needs the proper identification of the most responsible factor/factors for the bloom formation. However, in natural systems, algal blooms result from a combination of factors and from observation it is difficult to identify the most important one. In the present paper, using a mathematical model we compare the effects of three human induced factors (fertilizer input in agricultural field, eutrophication due to other sources than fertilizers, and overfishing) on the bloom dynamics and DO level. By applying a sophisticated sensitivity analysis technique, we found that the increasing use of fertilizers in agricultural field causes more rapid algal growth and decreases DO level much faster than eutrophication from other sources and overfishing. We also look at the mechanisms how fertilizer input rate affects the algal bloom dynamics and DO level. The model can be helpful for the policy makers in determining the influential factors responsible for the bloom formation.

A review of algal research in space

NASA Astrophysics Data System (ADS)

Niederwieser, Tobias; Kociolek, Patrick; Klaus, David

2018-05-01

With the continued expansion of human presence into space, typical mission durations will routinely exceed six months and extend to distances beyond the Moon. As such, sending periodic resupply vehicles, as currently provided to the International Space Station, will likely no longer be feasible. Instead, self-sustaining life support systems that recycle human waste products will become increasingly necessary, especially for planetary bases. The idea of bioregenerative life support systems using algal photobioreactors has been discussed since the beginning of the space age. In order to evaluate how such a system could be implemented, a variety of space flight studies aimed at characterizing the potential for using algae in air revitalization, water recycling, food production, and radiation shielding applications have been conducted over the years. Also, given the recent, growing interest in algal research for regenerative fuel production, food supplements, and cosmetics, many algal strains are already well documented from related terrestrial experiments. This paper reviews past algal experiments flown in space from 1960 until today. Experimental methods and results from 51 investigations utilizing either green algae (Chlorophyta), cyanobacteria (Cyanophyta), or Euglenophyta are analyzed and categorized by a variety of parameters, including size, species and duration. The collected data are summarized in a matrix that allows easy comparison between the experiments and provides important information for future life support system requirement definition and design. Similarities between experiment results are emphasized. Common problems and shortcomings are summarized and analyzed in terms of potential solutions. Finally, key research gaps, which must be closed before developing a functional life support system, are identified.

Gravity, chromosomes, and organized development in aseptically cultured plant cells

NASA Technical Reports Server (NTRS)

Krikorian, Abraham D.

1993-01-01

The objectives of the PCR experiment are: to test the hypothesis that microgravity will in fact affect the pattern and developmental progression of embryogenically competent plant cells from one well-defined, critical stage to another; to determine the effects of microgravity in growth and differentiation of embryogenic carrot cells grown in cell culture; to determine whether microgravity or the space environment fosters an instability of the differentiated state; and to determine whether mitosis and chromosome behavior are adversely affected by microgravity. The methods employed will consist of the following: special embryogenically competent carrot cell cultures will be grown in cell culture chambers provided by NASDA; four cell culture chambers will be used to grow cells in liquid medium; two dishes (plant cell culture dishes) will be used to grow cells on a semi-solid agar support; progression to later embryonic stages will be induced in space via crew intervention and by media manipulation in the case of liquid grown cell cultures; progression to later stages in case of semi-solid cultures will not need crew intervention; embryo stages will be fixed at a specific interval (day 6) in flight only in the case of liquid-grown cultures; and some living cells and somatic embryos will be returned for continued post-flight development and 'grown-out.' These will derive from the semi-solid grown cultures.

Expansion of Endothelial Progenitor Cells in High Density Dot Culture of Rat Bone Marrow Cells

PubMed Central

Wang, Ling; Kretlow, James D.; Zhou, Guangdong; Cao, Yilin; Liu, Wei; Zhang, Wen Jie

2014-01-01

In vitro expansion of endothelial progenitor cells (EPCs) remains a challenge in stem cell research and its application. We hypothesize that high density culture is able to expand EPCs from bone marrow by mimicking cell-cell interactions of the bone marrow niche. To test the hypothesis, rat bone marrow cells were either cultured in high density (2×105 cells/cm2) by seeding total 9×105 cells into six high density dots or cultured in regular density (1.6×104 cells/cm2) with the same total number of cells. Flow cytometric analyses of the cells cultured for 15 days showed that high density cells exhibited smaller cell size and higher levels of marker expression related to EPCs when compared to regular density cultured cells. Functionally, these cells exhibited strong angiogenic potentials with better tubal formation in vitro and potent rescue of mouse ischemic limbs in vivo with their integration into neo-capillary structure. Global gene chip and ELISA analyses revealed up-regulated gene expression of adhesion molecules and enhanced protein release of pro-angiogenic growth factors in high density cultured cells. In summary, high density cell culture promotes expansion of bone marrow contained EPCs that are able to enhance tissue angiogenesis via paracrine growth factors and direct differentiation into endothelial cells. PMID:25254487

Culture of Cells from Amphibian Embryos.

ERIC Educational Resources Information Center

Stanisstreet, Martin

1983-01-01

Describes a method for in vitro culturing of cells from amphibian early embryos. Such cells can be used to demonstrate such properties of eukaryote cells as cell motility, adhesion, differentiation, and cell sorting into tissues. The technique may be extended to investigate other factors. (Author/JN)

Preliminary identification of unicellular algal genus by using combined confocal resonance Raman spectroscopy with PCA and DPLS analysis

NASA Astrophysics Data System (ADS)

He, Shixuan; Xie, Wanyi; Zhang, Ping; Fang, Shaoxi; Li, Zhe; Tang, Peng; Gao, Xia; Guo, Jinsong; Tlili, Chaker; Wang, Deqiang

2018-02-01

The analysis of algae and dominant alga plays important roles in ecological and environmental fields since it can be used to forecast water bloom and control its potential deleterious effects. Herein, we combine in vivo confocal resonance Raman spectroscopy with multivariate analysis methods to preliminary identify the three algal genera in water blooms at unicellular scale. Statistical analysis of characteristic Raman peaks demonstrates that certain shifts and different normalized intensities, resulting from composition of different carotenoids, exist in Raman spectra of three algal cells. Principal component analysis (PCA) scores and corresponding loading weights show some differences from Raman spectral characteristics which are caused by vibrations of carotenoids in unicellular algae. Then, discriminant partial least squares (DPLS) classification method is used to verify the effectiveness of algal identification with confocal resonance Raman spectroscopy. Our results show that confocal resonance Raman spectroscopy combined with PCA and DPLS could handle the preliminary identification of dominant alga for forecasting and controlling of water blooms.

Cell-specific Labeling Enzymes for Analysis of Cell–Cell Communication in Continuous Co-culture*

PubMed Central

Tape, Christopher J.; Norrie, Ida C.; Worboys, Jonathan D.; Lim, Lindsay; Lauffenburger, Douglas A.; Jørgensen, Claus

2014-01-01

We report the orthologous screening, engineering, and optimization of amino acid conversion enzymes for cell-specific proteomic labeling. Intracellular endoplasmic-reticulum-anchored Mycobacterium tuberculosis diaminopimelate decarboxylase (DDCM.tub-KDEL) confers cell-specific meso-2,6-diaminopimelate-dependent proliferation to multiple eukaryotic cell types. Optimized lysine racemase (LyrM37-KDEL) supports D-lysine specific proliferation and efficient cell-specific isotopic labeling. When ectopically expressed in discrete cell types, these enzymes confer 90% cell-specific isotopic labeling efficiency after 10 days of co-culture. Moreover, DDCM.tub-KDEL and LyrM37-KDEL facilitate equally high cell-specific labeling fidelity without daily media exchange. Consequently, the reported novel enzyme pairing can be used to study cell-specific signaling in uninterrupted, continuous co-cultures. Demonstrating the importance of increased labeling stability for addressing novel biological questions, we compare the cell-specific phosphoproteome of fibroblasts in direct co-culture with epithelial tumor cells in both interrupted (daily media exchange) and continuous (no media exchange) co-cultures. This analysis identified multiple cell-specific phosphorylation sites specifically regulated in the continuous co-culture. Given their applicability to multiple cell types, continuous co-culture labeling fidelity, and suitability for long-term cell–cell phospho-signaling experiments, we propose DDCM.tub-KDEL and LyrM37-KDEL as excellent enzymes for cell-specific labeling with amino acid precursors. PMID:24820872

Competitive interaction between Ditylum Brightwellii and Skeletonema Costatum by toxic metabolites

NASA Astrophysics Data System (ADS)

Rijstenbil, J. W.

Comparative growth experiments were carried out in order to examine the role of toxic metabolites in the competition between two marine diatom species. Ditylum brightwellii and Skeletonema costatum exhibited mutual inhibition and auto-inhibition. Charcoal filtration did not entirely remove the toxicity. Algal extracts were more toxic than algal filtrates. Cell lysis induced by osmotic-shock treatment caused auto-inhibition in a dense culture of D. brightwellii; cells of this species recovered from a low salinity treatment after addition of charcoal to a culture. In mixed cultures the growth of both species may be affected by mutual inhibition. Toxicity of media depends on the growth phase of the competitors. In dense cultures, comparable with algal blooms in eutrophic waters, exocrines may be more effective than in diluted cultures ( cf. mesotrophic waters.) Substances excreted in dense blooms of S. costatum may inhibit competing species.

Compact Cell Settlers for Perfusion Cultures of Microbial (and Mammalian) Cells.

PubMed

Freeman, Cassandra A; Samuel, Premsingh S D; Kompala, Dhinakar S

2017-07-01

As microbial secretory expression systems have become well developed for microbial yeast cells, such as Saccharomyces cerevisiae and Pichia pastoris, it is advantageous to develop high cell density continuous perfusion cultures of microbial yeast cells to retain the live and productive yeast cells inside the perfusion bioreactor while removing the dead cells and cell debris along with the secreted product protein in the harvest stream. While the previously demonstrated inclined or lamellar settlers can be used for such perfusion bioreactors for microbial cells, the size and footprint requirements of such inefficiently scaled up devices can be quite large in comparison to the bioreactor size. Faced with this constraint, we have now developed novel, patent-pending compact cell settlers that can be used more efficiently with microbial perfusion bioreactors to achieve high cell densities and bioreactor productivities. Reproducible results from numerous month-long perfusion culture experiments using these devices attached to the 5 L perfusion bioreactor demonstrate very high cell densities due to substantial sedimentation of the larger live yeast cells which are returned to the bioreactor, while the harvest stream from the top of these cell settlers is a significantly clarified liquid, containing less than 30% and more typically less than 10% of the bioreactor cell concentration. Size of cells in the harvest is smaller than that of the cells in the bioreactor. Accumulated protein collected from the harvest and rate of protein accumulation is significantly (> 6x) higher than the protein produced in repeated fed-batch cultures over the same culture duration. © 2017 American Institute of Chemical Engineers Biotechnol. Prog., 33:913-922, 2017. © 2017 American Institute of Chemical Engineers.

Development of a microfluidic perfusion 3D cell culture system

NASA Astrophysics Data System (ADS)

Park, D. H.; Jeon, H. J.; Kim, M. J.; Nguyen, X. D.; Morten, K.; Go, J. S.

2018-04-01

Recently, 3-dimensional in vitro cell cultures have gained much attention in biomedical sciences because of the closer relevance between in vitro cell cultures and in vivo environments. This paper presents a microfluidic perfusion 3D cell culture system with consistent control of long-term culture conditions to mimic an in vivo microenvironment. It consists of two sudden expansion reservoirs to trap incoming air bubbles, gradient generators to provide a linear concentration, and microchannel mixers. Specifically, the air bubbles disturb a flow in the microfluidic channel resulting in the instability of the perfusion cell culture conditions. For long-term stable operation, the sudden expansion reservoir is designed to trap air bubbles by using buoyancy before they enter the culture system. The performance of the developed microfluidic perfusion 3D cell culture system was examined experimentally and compared with analytical results. Finally, it was applied to test the cytotoxicity of cells infected with Ewing’s sarcoma. Cell death was observed for different concentrations of H2O2. For future work, the developed microfluidic perfusion 3D cell culture system can be used to examine the behavior of cells treated with various drugs and concentrations for high-throughput drug screening.

An Introductory Undergraduate Course Covering Animal Cell Culture Techniques

ERIC Educational Resources Information Center

Mozdziak, Paul E.; Petitte, James N.; Carson, Susan D.

2004-01-01

Animal cell culture is a core laboratory technique in many molecular biology, developmental biology, and biotechnology laboratories. Cell culture is a relatively old technique that has been sparingly taught at the undergraduate level. The traditional methodology for acquiring cell culture training has been through trial and error, instruction when…

Cardiac Cells Beating in Culture: A Laboratory Exercise

ERIC Educational Resources Information Center

Weaver, Debora

2007-01-01

This article describes how to establish a primary tissue culture, where cells are taken directly from an organ of a living animal. Cardiac cells are taken from chick embryos and transferred to culture dishes. These cells are not transformed and therefore have a limited life span. However, the unique characteristics of cardiac cells are maintained…

Which has more stem-cell characteristics: Müller cells or Müller cells derived from in vivo culture in neurospheres?

PubMed

Ji, Hong-Pei; Xiong, Yu; Zhang, En-Dong; Song, Wei-Tao; Gao, Zhao-Lin; Yao, Fei; Sun, Hong; Zhou, Rong-Rong; Xia, Xiao-Bo

2017-01-01

Müller cells can be acquired from in vitro culture or a neurosphere culture system. Both culture methods yield cells with progenitor-cell characteristics that can differentiate into mature nervous cells. We compared the progenitor-cell traits of Müller cells acquired from each method. Primary murine Müller cells were isolated in serum culture media and used to generate Müller cells derived from neurospheres in serum-free culture conditions. Gene expression of neural progenitor cell markers was examined by Q-PCR in the two groups. Expression of rhodopsin and the cone-rod homeobox protein CRX were assessed after induction with 1 μM all-trans retinoic acid (RA) for 7 days. After more than four passages, many cells were large, flattened, and difficult to passage. A spontaneously immortalized Müller cell line was not established. Three-passage neurospheres yielded few new spheres. Genes coding for Nestin, Sox2, Chx10, and Vimentin were downregulated in cells derived from neurospheres compared to the cells from standard culture, while Pax6 was upregulated. Müller cells from both culture methods were induced into rod photoreceptors, but expression of rhodopsin and CRX was greater in the Müller cells from the standard culture. Both culture methods yielded cells with stem-cell characteristics that can be induced into rod photoreceptor neurons by RA. Serum had no influence on the "stemness" of the cells. Cells from standard culture had greater "stemness" than cells derived from neurospheres. The standard Müller cells would seem to be the best choice for transplantation in cell replacement therapy for photoreceptor degeneration.

Small herbivores suppress algal accumulation on Agatti atoll, Indian Ocean

NASA Astrophysics Data System (ADS)

Cernohorsky, Nicole H.; McClanahan, Timothy R.; Babu, Idrees; Horsák, Michal

2015-12-01

Despite large herbivorous fish being generally accepted as the main group responsible for preventing algal accumulation on coral reefs, few studies have experimentally examined the relative importance of herbivore size on algal communities. This study used exclusion cages with two different mesh sizes (1 × 1 cm and 6 × 6 cm) to investigate the impact of different-sized herbivores on algal accumulation rates on the shallow (<2 m) back-reef of Agatti atoll, Lakshadweep. The fine-mesh cages excluded all visible herbivores, which had rapid and lasting effects on the benthic communities, and, after 127 d of deployment, there was a visible and significant increase in algae (mainly macroalgae) with algal volume being 13 times greater than in adjacent open areas. The coarse-mesh cages excluded larger fishes (>8 cm body depth) while allowing smaller fishes to access the plots. In contrast to the conclusions of most previous studies, the exclusion of large herbivores had no significant effect on the accumulation of benthic algae and the amount of algae present within the coarse-mesh cages was relatively consistent throughout the experimental period (around 50 % coverage and 1-2 mm height). The difference in algal accumulation between the fine-mesh and coarse-mesh cages appears to be related to the actions of small individuals from 12 herbivorous fish species (0.17 ind. m-2 and 7.7 g m-2) that were able to enter through the coarse mesh. Although restricted to a single habitat, these results suggest that when present in sufficient densities and diversity, small herbivorous fishes can prevent the accumulation of algal biomass on coral reefs.

A Photosynthesis Lab. Response of Algal Suspensions to a Gradient of Photosynthetically Active Radiation (PAR).

ERIC Educational Resources Information Center

Zee, Delmar Vander

1995-01-01

This photosynthesis exercise is intended for introductory college biology or botany courses. It is based on the principle that a closed suspension of algal cells may be expected to produce more dissolved oxygen with a greater photon fluence rate, but within limits of the photosynthetic capacity of the system. Describes materials and methods. (LZ)

Advanced cell culture techniques for cancer drug discovery.

PubMed

Lovitt, Carrie J; Shelper, Todd B; Avery, Vicky M

2014-05-30

Human cancer cell lines are an integral part of drug discovery practices. However, modeling the complexity of cancer utilizing these cell lines on standard plastic substrata, does not accurately represent the tumor microenvironment. Research into developing advanced tumor cell culture models in a three-dimensional (3D) architecture that more prescisely characterizes the disease state have been undertaken by a number of laboratories around the world. These 3D cell culture models are particularly beneficial for investigating mechanistic processes and drug resistance in tumor cells. In addition, a range of molecular mechanisms deconstructed by studying cancer cells in 3D models suggest that tumor cells cultured in two-dimensional monolayer conditions do not respond to cancer therapeutics/compounds in a similar manner. Recent studies have demonstrated the potential of utilizing 3D cell culture models in drug discovery programs; however, it is evident that further research is required for the development of more complex models that incorporate the majority of the cellular and physical properties of a tumor.

Advanced Cell Culture Techniques for Cancer Drug Discovery

PubMed Central

Lovitt, Carrie J.; Shelper, Todd B.; Avery, Vicky M.

2014-01-01

Human cancer cell lines are an integral part of drug discovery practices. However, modeling the complexity of cancer utilizing these cell lines on standard plastic substrata, does not accurately represent the tumor microenvironment. Research into developing advanced tumor cell culture models in a three-dimensional (3D) architecture that more prescisely characterizes the disease state have been undertaken by a number of laboratories around the world. These 3D cell culture models are particularly beneficial for investigating mechanistic processes and drug resistance in tumor cells. In addition, a range of molecular mechanisms deconstructed by studying cancer cells in 3D models suggest that tumor cells cultured in two-dimensional monolayer conditions do not respond to cancer therapeutics/compounds in a similar manner. Recent studies have demonstrated the potential of utilizing 3D cell culture models in drug discovery programs; however, it is evident that further research is required for the development of more complex models that incorporate the majority of the cellular and physical properties of a tumor. PMID:24887773

Learning about Cells as Dynamic Entities: An Inquiry-Driven Cell Culture Project

ERIC Educational Resources Information Center

Palombi, Peggy Shadduck; Jagger, Kathleen Snell

2008-01-01

Using cultured fibroblast cells, undergraduate students explore cell division and the responses of cultured cells to a variety of environmental changes. The students learn new research techniques and carry out a self-designed experiment. Through this project, students enhance their creative approach to scientific inquiry, learn time-management and…

Dynamic cell culture system: a new cell cultivation instrument for biological experiments in space

NASA Technical Reports Server (NTRS)

Gmunder, F. K.; Nordau, C. G.; Tschopp, A.; Huber, B.; Cogoli, A.

1988-01-01

The prototype of a miniaturized cell cultivation instrument for animal cell culture experiments aboard Spacelab is presented (Dynamic cell culture system: DCCS). The cell chamber is completely filled and has a working volume of 200 microliters. Medium exchange is achieved with a self-powered osmotic pump (flowrate 1 microliter h-1). The reservoir volume of culture medium is 230 microliters. The system is neither mechanically stirred nor equipped with sensors. Hamster kidney (Hak) cells growing on Cytodex 3 microcarriers were used to test the biological performance of the DCCS. Growth characteristics in the DCCS, as judged by maximal cell density, glucose consumption, lactic acid secretion and pH, were similar to those in cell culture tubes.

Algal bloom-associated disease outbreaks among users of freshwater lakes-United States, 2009 - 2010

EPA Science Inventory

‘Algal blooms’ are local abundances of phytoplankton – microscopic photosynthesizing aquatic organisms found in surface waters worldwide; blooms are variable temporally and spatially and frequently produce a visible algal scum on the water. Harmful algal blooms (HABs) are abundan...

Modifications to the algal growth inhibition test for use as a regulatory assay

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

Geis, S.W.; Fleming, K.L.; Korthals, E.T.

2000-01-01

Biological assays using aquatic invertebrates and fish do not necessarily predict protection levels for primary producers such as algae and aquatic macrophytes. State regulatory programs may not be protecting the environment from many phytotoxic compounds. Recent modifications of the US Environmental Protection Agency's algal test were evaluated for their potential use as a regulatory assay. Primary goals of this investigation were to downsize the algal assay and to evaluate various methods of automation. Disposable microplates with 2-ml sample wells were evaluated as an alternative testing chamber for the 96-h growth inhibition test with Raphidocelis subcapitata (formerly known as Selenastrum capricornutum).more » The authors compared the standardized Erlenmeyer {reg_sign} flask test to the microplate test using CuCl{sub 2}, NaCl, phenol, ZnCl{sub 2}, and a surfactant. They noted improved control performance with the microplate test, whereas median inhibitory concentration values were similar for both methods. Other procedures they addressed included the use of EDTA, filtration of samples, and the effect of colored samples on algal growth. They also evaluated growth estimates by comparing manual cell counting to more automated growth estimates using fluorescence and absorbance endpoints. The use of fluorescence and absorbance measurements demonstrated reductions in replicate variability over manual counting and may offer time-saving alternatives for laboratory analysts.« less

Advancing Commercialization of Algal Biofuel through Increased Biomass Productivity and Technical Integration

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

Anton, David

The proposed project built on the foundation of over several years years of intensive and ground-breaking R&D work at Cellana's Kona Demonstration Facility (KDF). Phycological and engineering solutions were provided to tackle key cultivation issues and technical barriers limiting algal biomass productivity identified through work conducted outdoors at industrial (1 acre) scale. The objectives of this project were to significantly improve algal biomass productivity and reduce operational cost in a seawater-based system, using results obtained from two top-performing algal strains as the baseline while technically advancing and more importantly, integrating the various unit operations involved in algal biomass production, processing,more » and refining.« less

Three-dimensional Tissue Culture Based on Magnetic Cell Levitation

PubMed Central

Souza, Glauco R.; Molina, Jennifer R.; Raphael, Robert M.; Ozawa, Michael G.; Stark, Daniel J.; Levin, Carly S.; Bronk, Lawrence F.; Ananta, Jeyarama S.; Mandelin, Jami; Georgescu, Maria-Magdalena; Bankson, James A.; Gelovani, Juri G.

2015-01-01

Cell culture is an essential tool for drug discovery, tissue engineering, and stem cell research. Conventional tissue culture produces two-dimensional (2D) cell growth with gene expression, signaling, and morphology that can differ from those in vivo and thus compromise clinical relevancy1–5. Here we report a three-dimensional (3D) culture of cells based on magnetic levitation in the presence of hydrogels containing gold and magnetic iron oxide (MIO) nanoparticles plus filamentous bacteriophage. This methodology allows for control of cell mass geometry and guided, multicellular clustering of different cell types in co-culture through spatial variance of the magnetic field. Moreover, magnetic levitation of human glioblastoma cells demonstrates similar protein expression profiles to those observed in human tumor xenografts. Taken together, these results suggest levitated 3D culture with magnetized phage-based hydrogels more closely recapitulates in vivo protein expression and allows for long-term multi-cellular studies. PMID:20228788

The expression of native and cultured human retinal pigment epithelial cells grown in different culture conditions.

PubMed

Tian, J; Ishibashi, K; Honda, S; Boylan, S A; Hjelmeland, L M; Handa, J T

2005-11-01

To determine the transcriptional proximity of retinal pigment epithelium (RPE) cells grown under different culture conditions and native RPE. ARPE-19 cells were grown under five conditions in 10% CO(2): "subconfluent" in DMEM/F12+10% FBS, "confluent" in serum and serum withdrawn, and "differentiated" for 2.5 months in serum and serum withdrawn medium. Native RPE was laser microdissected. Total RNA was extracted, reverse transcribed, and radiolabelled probes were hybridised to an array containing 5,353 genes. Arrays were evaluated by hierarchical cluster analysis and significance analysis of microarrays. 78% of genes were expressed by native RPE while 45.3--47.7% were expressed by ARPE-19 cells, depending on culture condition. While the most abundant genes were expressed by native and cultured cells, significant differences in low abundance genes were seen. Hierarchical cluster analysis showed that confluent and differentiated, serum withdrawn cultures clustered closest to native RPE, and that serum segregated cultured cells from native RPE. The number of differentially expressed genes and their function, and profile of expressed and unexpressed genes, demonstrate differences between native and cultured cells. While ARPE-19 cells have significant value for studying RPE behaviour, investigators must be aware of how culture conditions can influence the mRNA phenotype of the cell.

Evaluation of the osteogenic differentiation of gingiva-derived stem cells grown on culture plates or in stem cell spheroids: Comparison of two- and three-dimensional cultures.

PubMed

Lee, Sung-Il; Ko, Youngkyung; Park, Jun-Beom

2017-09-01

Three-dimensional cell culture systems provide a convenient in vitro model for the study of complex cell-cell and cell-matrix interactions in the absence of exogenous substrates. The current study aimed to evaluate the osteogenic differentiation potential of gingiva-derived stem cells cultured in two-dimensional or three-dimensional systems. To the best of our knowledge, the present study is the first to compare the growth of gingiva-derived stem cells in monolayer culture to a three-dimensional culture system with microwells. For three-dimensional culture, gingiva-derived stem cells were isolated and seeded into polydimethylsiloxane-based concave micromolds. Alkaline phosphatase activity and alizarin red S staining assays were then performed to evaluate osteogenesis and the degree of mineralization, respectively. Stem cell spheroids had a significantly increased level of alkaline phosphatase activity and mineralization compared with cells from the two-dimensional culture. In addition, an increase in mineralized deposits was observed with an increase in the loading cell number. The results of present study indicate that gingiva-derived stem cell spheroids exhibit an increased osteogenic potential compared with stem cells from two-dimensional culture. This highlights the potential of three-dimensional culture systems using gingiva-derived stem cells for regenerative medicine applications requiring stem cells with osteogenic potential.

Fundamentals of microfluidic cell culture in controlled microenvironments†

PubMed Central

Young, Edmond W. K.; Beebe, David J.

2010-01-01

Microfluidics has the potential to revolutionize the way we approach cell biology research. The dimensions of microfluidic channels are well suited to the physical scale of biological cells, and the many advantages of microfluidics make it an attractive platform for new techniques in biology. One of the key benefits of microfluidics for basic biology is the ability to control parameters of the cell microenvironment at relevant length and time scales. Considerable progress has been made in the design and use of novel microfluidic devices for culturing cells and for subsequent treatment and analysis. With the recent pace of scientific discovery, it is becoming increasingly important to evaluate existing tools and techniques, and to synthesize fundamental concepts that would further improve the efficiency of biological research at the microscale. This tutorial review integrates fundamental principles from cell biology and local microenvironments with cell culture techniques and concepts in microfluidics. Culturing cells in microscale environments requires knowledge of multiple disciplines including physics, biochemistry, and engineering. We discuss basic concepts related to the physical and biochemical microenvironments of the cell, physicochemical properties of that microenvironment, cell culture techniques, and practical knowledge of microfluidic device design and operation. We also discuss the most recent advances in microfluidic cell culture and their implications on the future of the field. The goal is to guide new and interested researchers to the important areas and challenges facing the scientific community as we strive toward full integration of microfluidics with biology. PMID:20179823

A novel three-dimensional cell culture method enhances antiviral drug screening in primary human cells.

PubMed

Koban, Robert; Neumann, Markus; Daugs, Aila; Bloch, Oliver; Nitsche, Andreas; Langhammer, Stefan; Ellerbrok, Heinz

2018-02-01

Gefitinib is a specific inhibitor of the epidermal growth factor receptor (EGFR) and FDA approved for treatment of non-small cell lung cancer. In a previous study we could show the in vitro efficacy of gefitinib for treatment of poxvirus infections in monolayer (2D) cultivated cell lines. Permanent cell lines and 2D cultures, however, are known to be rather unphysiological; therefore it is difficult to predict whether determined effective concentrations or the drug efficacy per se are transferable to the in vivo situation. 3D cell cultures, which meanwhile are widely distributed across all fields of research, are a promising tool for more predictive in vitro investigations of antiviral compounds. In this study the spreading of cowpox virus and the antiviral efficacy of gefitinib were analyzed in primary human keratinocytes (NHEK) grown in a novel 3D extracellular matrix-based cell culture model and compared to the respective monolayer culture. 3D-cultivated NHEK grew in a polarized and thus a more physiological manner with altered morphology and close cell-cell contact. Infected cultures showed a strongly elevated sensitivity towards gefitinib. EGFR phosphorylation, cell proliferation, and virus replication were significantly reduced in 3D cultures at gefitinib concentrations which were at least 100-fold lower than those in monolayer cultures and well below the level of cytotoxicity. Our newly established 3D cell culture model with primary human cells is an easy-to-handle alternative to conventional monolayer cell cultures and previously described more complex 3D cell culture systems. It can easily be adapted to other cell types and a broad spectrum of viruses for antiviral drug screening and many other aspects of virus research under more in vivo-like conditions. In consequence, it may contribute to a more targeted realization of necessary in vivo experiments. Copyright © 2017 Elsevier B.V. All rights reserved.

Development of suspension cell culture model to mimic circulating tumor cells

PubMed Central

Park, Ji Young; Jeong, Ae Lee; Joo, Hyun Jeong; Han, Sora; Kim, So-Hyun; Kim, Hye-Youn; Lim, Jong-Seok; Lee, Myeong-Sok; Choi, Hyung-Kyoon; Yang, Young

2018-01-01

Circulating tumor cells (CTCs) are essential for the establishment of distant metastasis. Numerous studies have characterized CTCs as metastatic precursors; however, the molecular nature of CTCs has not been completely revealed yet due to the low number of CTCs in the blood stream. As an alternative approach, we developed a long-term suspension cell culture model using human breast cancer cell lines to mimic CTCs. We found that more than 40 passaged suspension cells acquired the ability to enhance metastasis like cancer stem cells. To identify molecular changes acquired during the suspension cell culture, we analyzed metabolic and lipidomic profiles as well as transcriptome in MDA-MB-468 suspension cells. Glutamate and leucine levels increased in suspension cells, and cholesterol synthesis pathway was altered. The inhibition of glutamate metabolic pathway decreased the proliferation of suspension cells compared to that of adherent cells. In the lipidomic profile, PC species containing long chain and polyunsaturated fatty acids increased in suspension cells and these species could be authentic and specific biomarkers for highly metastatic cancers. As this CTC-mimicking suspension cell culture model may easily apply to various types of cancer, we suggest this model as a great tool to develop therapeutic targets and drugs to eradicate metastatic cancer cells. PMID:29416640

Magnetic 3D Cell Culturing

NASA Image and Video Library

2017-07-11

iss052e014201 (7/11/2017) --- NASA astronaut Peggy Whitson uses a microscope to view Magnetic 3D Biocells. This investigation uses magnetized cells and tools to make it easier to handle cells and cultures and to improve the reproducibility of experiments.

Primary Human Uterine Leiomyoma Cell Culture Quality Control: Some Properties of Myometrial Cells Cultured under Serum Deprivation Conditions in the Presence of Ovarian Steroids.

PubMed

Bonazza, Camila; Andrade, Sheila Siqueira; Sumikawa, Joana Tomomi; Batista, Fabrício Pereira; Paredes-Gamero, Edgar J; Girão, Manoel J B C; Oliva, Maria Luiza V; Castro, Rodrigo Aquino

2016-01-01

Cell culture is considered the standard media used in research to emulate the in vivo cell environment. Crucial in vivo experiments cannot be conducted in humans and depend on in vitro methodologies such as cell culture systems. However, some procedures involving the quality control of cells in culture have been gradually neglected by failing to acknowledge that primary cells and cell lines change over time in culture. Thus, we report methods based on our experience for monitoring primary cell culture of human myometrial cells derived from uterine leiomyoma. We standardized the best procedure of tissue dissociation required for the study of multiple genetic marker systems that include species-specific antigens, expression of myofibroblast or myoblast markers, growth curve, serum deprivation, starvation by cell cycle synchronization, culture on collagen coated plates, and 17 β-estradiol (E2) and progesterone (P4) effects. The results showed that primary myometrial cells from patients with uterine leiomyoma displayed myoblast phenotypes before and after in vitro cultivation, and leiomyoma cells differentiated into mature myocyte cells under the appropriate differentiation-inducing conditions (serum deprivation). These cells grew well on collagen coated plates and responded to E2 and P4, which may drive myometrial and leiomyoma cells to proliferate and adhere into a focal adhesion complex involvement in a paracrine manner. The establishment of these techniques as routine procedures will improve the understanding of the myometrial physiology and pathogenesis of myometrium-derived diseases such as leiomyoma. Mimicking the in vivo environment of fibrotic conditions can prevent false results and enhance results that are based on cell culture integrity.