Biomass and nutrient productivities ofTetraselmis chuii under mixotrophic culture conditions with various C：N ratios

Mass microalgal culture plays an irreplaceable role in aquaculture,but microalgal productivity is restricted by traditional autotrophic culture conditions.In the present study,a Tetraselmis chuii strain belonging to the phylum Chlorophyta was isolated from south Yellow Sea.The growth rate and biomass productivity of this strain was higher under mixotrophic conditions with different carbon:nitrogen(C:N)ratios than those under autotrophic conditions.When the C:N ratio was 16,the optical density and biomass productivity were 3.7-and 5-fold higher than their corresponding values under autotrophic culture conditions,respectively.Moreover,T.chuii synthesized more polysaccharides and total lipids under mixotrophic conditions.In addition,T.chuii cultured under mixotrophic conditions synthesized more types of fatty acids than autotrophic culture conditions.At a C:N ratio of 16,the percentage of C16:0 and C18:1 reached 30.08%and 24.65%of the total fatty acid(TFA) content,respectively.These findings may provide a basis for largescale mixotrophic culture of T.chuii,as a potential bait-microalga.

Growth and Physiological Features of Cyanobacterium Anabaena sp. Strain PCC 7120 in a Glucose-Mixotrophic Culture

Mixotrophic growth is one potential mode for mass culture of microalgae and cyanobacteria particularly suitable for the production of high value bioactive compounds and fine chemicals.The typical heterocystous cyanobacterium Anabaena sp.PCC 7120 was grown in the presence of exogenous glucose in light.Glucose improved the cell growth evidently,the maximal specific growth rate under mixotrophic condition（0.38 d 1）being 1.6-fold of that of photoautotrophic growth.Mixotrophy caused a variation in cellular pigment composition,increasing the content of chlorophyll a and decreasing the contents of carotenoid and phycobiliprotein relative to chlorophyll a.Fluorescence emission from photosystem II（PSII）relative to photosystem I was enhanced in mixotrophic cells,implying an increased energy distribution in PSII.Glucokinase（EC 2.7.1.2）activity was further induced in the presence of glucose.The mixotrophic culture was scaled up in a 15 L airlift photobioreactor equipped with an inner and an outer light source.A modified Monod model incorporating the specific growth rate and the average light intensity in the reactor was developed to describe cell growth appropriately.The understanding of mixotrophic growth and relevant physiological features of Anabaena sp.PCC 7120 would be meaningful for cultivation and exploitation of this important cyanobacterial strain.

Mixotrophic Cultivation of Tetraselmis sp.-1 in Airlift Photobioreactor

Tetraselmis sp.-1 is a new microalgae strain constructed by cell fusion technique. In this paper, the mixotrophic cultivation of Tetraselmis sp.-1 in airlift photobioreactor is investigated. Firstly, the paper calculates the light attenuation in the mixotrophic medium, and sets the light attenuation model. Secondly, it uses the same dissolved oxygen coefficient (K d) of flask culture to select the aeration of bioreactor. Finally, it sets the growth kinetic model, production (chlorophyll-a and total lipid) kinetic models and substrate (glucose) consumption kinetic model of Tetraselmis sp.-1 in airlift photobioreactor.

Degeneration of photosynthetic capacity in mixotrophic plants, Chimaphila japonica and Pyrola decorata (Ericaceae)

The evolution of photosynthesis is an important feature of mixotrophic plants.Previous inferences proposed that mixotrophic taxa tend to retain most genes relating to photosynthetic functions but vary in plastid gene content.However,no sequence data are available to test this hypothesis in Ericaceae.To investigate changes in plastid genomes that may result from a transition from autotrophy to mixotrophy,the plastomes of two mixotrophic plants,Pyrola decorata and Chimaphila japonica,were sequenced at Illumina's Genome Analyzer and compared to the published plastome of the autotrophic plant Rhododendron simsii,which also belongs to Ericaceae.The greatest discrepancy between mixotrophic and autotrophic plants was that ndh genes for both P.decorata and C.japonica plastomes have nearly all become pseudogenes.P.decorata and C.japonica also retained all genes directly involved in photosynthesis under strong selection.The calculated rate of nonsynonymous nucleotide substitutions and synonymous substitutions of protein-coding genes(dN/dS) showed that substitution rates in shade plants were apparently higher than those in sunlight plants.The two mixotrophic plastomes were generally very similar to that of non-parasitic plants,although ndh genes were largely pseudogenized.Photosynthesis genes under strong selection were retained in the two mixotrophs,however,with greatly increased substitution rates.Further research is needed to gain a clearer understanding of the evolution of autotrophy and mixotrophy in Ericaceae.

Woodchip-sulfur packed biological permeable reactive barrier for mixotrophic vanadium(V)detoxification in groundwater

Groundwater vanadium(V)(V(V))contamination is ubiquitous in vanadium mining/smelting region and development of novel strategy for its remediation is of particular significance.Herein woodchip-sulfur packed biological permeable reactive barrier(bio-PRB)is established towards successful V(V)bio-detoxification.V(V)removal was accelerated under such mixotrophic condition,compared with heterotrophic and autotrophic V(V)reductions.The performance of bio-PRB was relatively steady with V(V)removal efficiency of 68.5%–98.2%under fluctuant geochemical and hydrodynamic conditions.Microbial community analysis indicated that heterotrophic Geobacter was the main reducer to convert V(V)to insoluble V(IV),by consumption of organic source attributed to woodchip hydrolysis and sulfur anabolism of autotrophs(e.g.,Sulfuricurvum and Thiobacillus).V(V)reduction and elemental sulfur oxidation were regulated by genes as omcA,omcB and mtrC and soxB,respectively.The elevated contents of cytochrome c and nicotinamide adenine dinucleotide implied that improved electron transfer facilitated V(V)reduction.This study provides a cost-effective,robust and sustainable route for V(V)-polluted aquifer remediation.

Microalgal bioremediation of food-processing industrial wastewater under mixotrophic conditions： Kinetics and scaleup approach

The Chlorella microalgae were mixotrophically cultivated in an unsterilized and unfikered raw foodprocessing industrial wastewater. Both inorganic carbon （CO2-air） and organic carbon （wastewater） were provided simultaneously for microalgae growth. The aim of the study is to find out the utilization rates of total organic carbon （TOC） and chemical oxygen demand （COD） under mixotrophic conditions for a given waste water. About 90% reduction in TOC and COD were obtained for all dilutions of wastewater. Over 60% of nitrate and 40% of phosphate were consumed by microalgae from concentrated raw wastewater. This study shows that microalgae can use both organic and inorganic sources of carbon in more or less quantity under mixotrophic conditions. The growth of microalgae in food-processing industrial wastewater with all studied dilution factors, viz. zero （raw）, 1.6 （dilution A）, and 5 （dilution B） suggests that the freshwater requirement could be reduced substantially （20%-60%）. The degradation kinetics also suggests that the microalgae cultivation on a high COD wastewater is feasible and scalable.

Isolation and characterization of facultative mixotrophic ammonia-oxidizing bacteria from constructed wetlands

Autotrophic ammonia-oxidizing bacteria （AOB） have been widely studied in constructed wetlands systems, while mixotrophic AOB have been less thoroughly examined. Heterotrophic bacteria were isolated from wastewater and rhizospheres of macrophytes of constructed wetlands, and then cultivated in a mixotrophic medium containing ammonium and acetic acid. A molecular characterization was accomplished using ITS-PCR amplification, and phylogenetic analysis based on 16S rRNA gene Sequences. Results showed the presence of 35 bacteria, among 400 initially heterotrophic isolates, that were able to remove ammonia. These 35 isolates were classified into 10 genetically different groups based on ITS pattern. Then, a collection of 10 isolates were selected because of their relatively high ammonia removal efficiencies （ARE≥ 80%） and their phylogenetic diversity. In conditions of mixotrophy, these strains were shown to be able to grow （increase of optical density OD660 during incubation with assimilation of nitrogen into cellular biomass） and to oxidize ammonia （important ammonia oxidation efficiencies, AOE between 79% and 87%）. Among these facultative mixotrophic AOB, four isolates were genetically related to Firmicutes （Bacillus and Exiguobacterium）, three isolates were affiliated to Actinobacteria （Arthrobacter） and three other isolates were associated with Proteobacteria （Pseudomonas, Ochrobactrum and Bordetella）.

Thiosulfate oxidation and autotrophy potential by marine prevalent heterotrophic bacteria of genus Marinobacter

The genus Marinobacter is very broadly distributed in global environments and is considered as aerobic heterotroph.In this study,six Marinobacter strains were identified with autotrophic thiosulfate oxidation capacity.These strains,namely Marinobacter guineae M3B^(T),Marinobacter aromaticivorans D15-8PT,Marinobacter vulgaris F01^(T),Marinobacter profundi PWS21^(T),Marinobacter denitrificans JB02H27T,and Marinobacter sp.ST-1M(with a 99.93%similarity to the 16S rDNA sequences of Marinobacter salsuginis SD-14B^(T)),were screened out of 32 Marinobacter strains by autotrophic thiosulfate oxidization medium.The population of cells grew in a chemolithotrophic medium,increasing from 105 cells/mL to 10^(7) cells/mL within 5 d.This growth was accompanied by the consumption of thiosulfate 3.59 mmol/L to 9.64 mmol/L and the accumulation of sulfate up to 0.96 mmol/L,and occasionally produced sulfur containing complex particles.Among these Marinobacter strains,it was also found their capability of oxidizing thiosulfate to sulfate in a heterotrophic medium.Notably,M.vulgaris F01^(T)and M.antarcticus ZS2-30^(T)showed highly significant production of sulfate at 9.45 mmol/L and 3.10 mmol/L.Genome annotation indicated that these Marinobacter strains possess a complete Sox cluster for thiosulfate oxidation.Further phylogenetic analysis of the soxB gene revealed that six Marinobacter strains formed a separate lineage within Gammaproteobacteria and close to obligate chemolithoautotroph Thiomicrorhabdus arctica.The results indicated that thiosulfate oxidizing and chemolithoautotrophic potential in Marinobacter genus,which may contribute to the widespread of Marinobacter in the global ocean.

Unraveling the role of formate in improving nitrogen removal via coupled partial denitrification-anammox

The addition of traditional carbon sources(e.g.,acetate)could favor heterotrophic overgrowth in partial denitrification coupled with anammox(PD–A)systems,thus hindering the performance and stability of this novel wastewater nitrogen removal technology.Therefore,it is necessary to develop an effective,environmentally friendly,and inexpensive alternative.This study demonstrated the potential of formate to enhance the performance and community stability of PD–A under mainstream conditions.In a laboratory-scale biofilm reactor,formate addition(COD/NO_(3)^(–)–N=1.75)improved nitrogen removal efficiency(from 72.1±3.5%to 81.7±2.7%),EPS content(from 106.3±8.1 to 163.0±15.5 mg/gVSS)and increased anammox bacteria growth(predominantly Candidatus Brocadia,from 29.5±0.7%to 34.5±5.4%)while maintaining stable heterotrophs dominated by methylotrophic Desulfobacillus.FISH–NanoSIMS revealed a formate uptake using Ca.Brocadia and Desulfobacillus,with Ca.Brocadia being the major contributor to partial nitrate reduction to nitrite.Desulfobacillus can synthesize diverse hydrophobic amino acids and provide key nutrients for Ca.Brocadia.To achieve comparable nitrogen removal,the cost of the formate-driven PD–A process should be 11.2%lower than that of acetate.These results greatly enrich our understanding of C1 metabolism represented by formate in anammox communities and its application in the context of coupling partial denitrification–anammox toward enhanced nitrogen removal in global wastewater treatment systems.

Advances and challenges of sulfur-driven autotrophic denitrification(SDAD)for nitrogen removal

Sulfur-driven autotrophic denitrification(SDAD),a process suited for the treatment of nitrogen and sulfur-polluted wastewater without extra supplement of organic carbon,is a promising biological nitrogen removal process.However,the SDAD process was affected by many factors such as various electron donors,organic carbon and exogenous substances(e.g.,antibiotics and heavy metal),which prevent further application.Thus,we conducted a detailed review of previous studies on such influence factors and its current application.Besides,a comparative analysis was adopted to recognize the current challenges and future needs for feasible application,so as to ultimately perfect the SDAD process and extend its application scope.

Effects of Glycerol on the Fluorescence Spectra and Chloroplast Ultrastructure of Phaeodactylum tricornutum(Bacillariophyta)

Responses of the photosynthetic activity of Phaeodactylum tricornutum （Bacillariophyta） to organic carbon glycerol were investigated. The growth rate, photosynthetic pigments, 77 K fluorescence spectra, and chloroplast ultrastructure of P. tricornutum were examined under photoautotrophic, mixotrophic, and photoheterotrophic conditions. The results showed that the specific growth rate was the fastest under mixotrophic conditions. The cell photosynthetic pigment content and values of Chl a/Chl c were reduced under mixotrophic and photoheterotrophic conditions. The value of carotenoid/Chl a was enhanced under mixotrophic conditions, but was decreased under photoheterotrophic conditions. In comparison with photoautotrophic conditions, the fluorescence emission peaks and fluorescence excitation peaks were not shifted. The relative fluorescence of photosystem （PS） Ⅰ and PS Ⅱ and the values of F685/F710 and F685/F738 were decreased. Chloroplast thylakoid pairs were less packed under mixotrophic and photoheterotrophic conditions. There was a strong correlation between degree of chloroplast thylakoid packing and the excitation energy kept in PS Ⅱ. These results suggested that the PS Ⅱ activity was reduced by glycerol under mixotrophic conditions, thereby leading to repression of the photosynthetic activity.