Algal-bacterial Co-cultures Research Articles

Influence of growth medium on the species-specific interactions between algae and bacteria.

In this study, we investigated a species-specific algal-bacterial co-culture that has recently attracted worldwide scientific attention as a novel approach to enhancing algal growth rate. We report that the type of interaction between Chlorella vulgaris and bacteria of the genus Delftia is not solely determined by species specificity. Rather, it is a dynamic process of adaptation to the surrounding conditions, where one or the other microorganism dominates (temporally) depending on the growth conditions, in particular the medium. Under laboratory conditions, we found that Delftia sp. had a negative effect on C. vulgaris growth when co-cultured in a TAP medium. However, the co-culture of algae and bacteria under BG-11 and BG-11 + acetic acid resulted in an increase in algal concentration compared to algal cultures without bacteria under the same conditions. Additional chemical analysis revealed that the presence of different carbon (the main organic carbon source-acetic acid in TAP or BG-11 + acetic acid medium and inorganic carbon source-Na2CO3 in BG-11 or BG-11 + acetic acid medium) and nitrogen (NH4Cl in TAP medium and NaNO3 in BG-11 or BG-11 + acetic acid medium) species in the growth medium was one of the main factors driving the shift in interaction type.

Utilization and transformation of Chrysotila dentata-derived dissolved organic matter by phycosphere bacteria Marinobacter hydrocarbonoclasticus and Bacillus firmus.

The dissolved organic matter (DOM) released from the cocoolithophores (Chrysotila dentata) was studied in laboratory experiments after co-culturing C. dentata with bacteria. Marinobacter hydrocarbonoclasticus (CA6)-γ-Proteobacteria and Bacillus firmus (CF2) were used to investigate the utilization and processing of the DOM derived from C. dentata, utilizing fluorescence excitation-emission matrix (EEM) combined with parallel factor analysis (EEM-PARAFAC), while measuring algal abundance and photosynthetic parameters. The experimental groups consisted of axenic C. dentata groups, filter cultured with bacteria (CA6 or CF2) groups, C. dentata co-cultured with bacteria (CA6 or CF2) groups and axenic bacteria (CA6 or CF2) groups. We then evaluated the processing of DOM by determining four fluorescence indices. The number of C. dentata cells and the photosynthetic capacity of microalgae were enhanced by CA6 and CF2. The main known fluorophores, including humic-like components and protein-like components, were present in all sample. The protein-like component of algal-bacterial co-cultures was effectively utilized by CA6 and CF2. The humic-like components increased at the end of the culture time for all cultures. Meanwhile, the average fluorescence intensity of protein-like in CA6 co-culture with algae was lower than that in CF2 co-culture with algae over time. On the other hand, the average fluorescence intensity of humic-like in CA6 was higher than CF2. However, the total change in fluorescence in humic-like and protein-like of axenic CF2 cultures was lower than that of CA6. Hence, the ability of CA6 to transform microalgal-derived DOM was superior to that of CF2, and CF2's ability to consume bacterial-derived DOM was superior to that of CA6.

Algal Hydrogen Production and Exopolysaccharide Patterns in Chlorella–Bacillus Inter-Kingdom Co-Cultures

Biohydrogen production from wastewater using eukaryotic green algae can be facilitated by appropriately selected bacterial partners and cultivation conditions. Two Chlorella algal species were chosen for these experiments, based on their robust growth ability in synthetic wastewater. The applied three Bacillus bacterial partners showed active respiration and efficient biomass production in the same synthetic wastewater. Bacillus amyloliquefaciens, Bacillus mycoides, and Bacillus cereus as bacterial partners were shown to specifically promote algal biomass yield. Various inter-kingdom co-culture combinations were investigated for algal–bacterial biomass generation, for co-culture-specific exopolysaccharide patterns, and, primarily, for algal biohydrogen evolution. Chlorella sp. MACC-38 mono- and co-cultures generated significantly higher biomass compared with that of Chlorella sp. MACC-360 mono- and co-cultures, while in terms of hydrogen production, Chlorella sp. MACC-360 co-cultures clearly surpassed their Chlorella sp. MACC-38 counterparts. Imaging studies revealed tight physical interactions between the algal and bacterial partners and revealed the formation of co-culture-specific exopolysaccharides. Efficient bacterial respiration was in clear correlation with algal hydrogen production. Stable and sustainable algal hydrogen production was observed in synthetic wastewater for Chlorella sp. MACC-360 green algae in co-cultures with either Bacillus amyloliquefaciens or Bacillus cereus. The highest algal hydrogen yields (30 mL H2 L−1 d−1) were obtained when Chlorella sp. MACC-360 was co-cultured with Bacillus amyloliquefaciens. Further co-culture-specific algal biomolecules such as co-cultivation-specific exopolysaccharides increase the valorization potential of algal–bacterial co-cultures and might contribute to the feasibility of algal biohydrogen production technologies.

Interaction of Scenedesmus quadricauda and native bacteria in marine biopharmaceutical wastewater for desirable lipid production and wastewater treatment

Improving knowledge of the alga–bacterium interaction can promote the wastewater treatment. The untreated marine biopharmaceutical wastewater (containing native bacteria) was used directly for culturing microalgae. Unlike previous studies on specific bacteria in algal–bacterial co-culture systems, the effect of native bacteria in wastewater on microalgae growth was investigated in this study. The results showed that the coexistence of native bacteria greatly promoted the microalgae growth, ultimately producing biomass of 0.64 g/L and biomass productivity of 56.18 mg/L·d. Moreover, the lipid accumulation in the algae + bacteria group was 1.31 and 1.13 times higher than those of BG11 and pure algae, respectively, mainly attributed to the fact that bacteria provided a good environment for microalgae growth by using extracellular substances released from microalgae for their own growth, and providing micromolecules of organic matter and other required elements to microalgae. This study would lay the theoretical foundation for improving biopharmaceutical wastewater treatment.

Global metabolome analysis of Dunaliella tertiolecta, Phaeobacter italicus R11 Co-cultures using thermal desorption - Comprehensive two-dimensional gas chromatography - Time-of-flight mass spectrometry (TD-GC×GC-TOFMS)

Dunaliella tertiolecta is a marine microalgae that has been studied extensively as a potential carbon-neutral biofuel source (Tang et al., 2011). Microalgae oil contains high quantities of energy-rich fatty acids and lipids, but is not yet commercially viable as an alternative fuel. Carefully optimised growth conditions, and more recently, algal-bacterial co-cultures have been explored as a way of improving the yield of D. tertiolecta microalgae oils. The relationship between the host microalgae and bacterial co-cultures is currently poorly understood. Here, a complete workflow is proposed to analyse the global metabolomic profile of co-cultured D. tertiolectra and Phaeobacter italicus R11, which will enable researchers to explore the chemical nature of this relationship in more detail. To the best of the authors’ knowledge this study is one of the first of its kind, in which a pipeline for an entirely untargeted analysis of the algal metabolome is proposed using a practical sample preparation, introduction, and data analysis routine.

Microalgal–Bacterial Synergistic Interactions and Their Potential Influence in Wastewater Treatment: a Review

Microalgae, as the most promising raw material for biodiesel, can absorb and transform N, P, and organic matter in wastewater into cell components such as oil, carbohydrate, and protein. The cultivation of microalgae in wastewater provides the possibility to realize harmless treatment and resource utilization of wastewater and reduce the cost of microalgal culture. Whether in a single microalgal culture process or wastewater system, there always be a large number of bacteria interacting with microalgae, forming a complex microecological system. Although microalgal–bacterial coculture system is feasible features for the higher biomass production and better nutrient adaptability from wastewater, the precise understanding of interaction and coexistence in water bodies is still unclear. Some major challenges are to develop sustainability in the microalgal–bacterial coculture system due to the incomplete knowledge about the mechanism of communication, algal growth promotion, and effect of microalgae on the indigenous bacteria in wastewater. In the present review, nutritional interaction and signal transduction, two main interaction models of algae and bacteria, and the effect of synergism of these two models on microalgal system are summarized. Moreover, the impact of algal–bacterial coculture on algal growth and removal of pollutants in wastewater treatment process was expounded systematically to develop highly efficient consortium systems by covering the sustainability limitations. This work would provide guidance for the establishment of efficient algal–bacterial symbiosis system in the wastewater with complex microecology and nutrient environment to improve the accumulation of microalgal biomass and reduce the cost of microalgal culture.

Growth performance and nutrient removal of a Chlorella vulgaris-Rhizobium sp. co-culture during mixotrophic feed-batch cultivation in synthetic wastewater

The subarctic green algal strain Chlorella vulgaris 13-1, newly isolated from Northern Sweden, and its co-occurring bacterium Rhizobium sp. were tested for their ability to produce valuable biomass and remove nutrients from synthetic wastewater during mixotrophic feed-batch cultivation at multiple hydraulic retention times (HRTs=7, 5 and 3 days). The algal-bacterial co-culture showed better performance compared to the corresponding axenic cultures (HRT=7 days), with a biomass concentration of 0.63±0.03g/L and removal rates of 49.5±6.1% TOC, 55.7±8.04% TN and 95.6±3.6% TP at steady-state. Culture stability and a high nutrient removal capacity were recorded in the algal-bacterial co-culture even at HRTs of 5 and 3 days. Interestingly, reducing the HRT from 7 to 5 days resulted in a higher lipid content of the biomass, further reduction of the HRT to 3 days enhanced both daily biomass productivity (1.03g/L/day) and nutrient assimilation. Mixotrophic co-cultivation of C. vulgaris-Rhizobium sp. can successfully be applied for wastewater reclamation in continuous mode at HRT of 3–7 days, and thus is suitable for both summer and winter conditions in Nordic countries.

Bacterial communities protect the alga Microchloropsis salina from grazing by the rotifer Brachionus plicatilis

Open algal ponds are likely to succumb to unpredictable, devastating crashes by one or several deleterious species. Developing methodology to mitigate or prevent pond crashes will increase algal biomass production, drive down costs for algae farmers, and reduce the risk involved with algae cultivation, making it more favorable for investment by entrepreneurs and biotechnology companies. Here, we show that specific algal-bacterial co-cultures grown with the green alga Microchloropsis salina prevented grazing by the marine rotifer, Brachionus plicatilis. We obtained seven algal-bacterial co-cultures from crashed rotifer cultures, maintained them in co-culture with Microchloropsis salina, and used a microalgal survival assay to determine that algae present in each co-culture were protected from rotifer grazing and culture crash. After months of routinely diluting and maintaining these seven algal-bacterial co-cultures, we repeated the assay and found the opposite result: none of the seven bacterial communities protected the microalgae from rotifer grazing. We performed 16S rRNA gene amplicon sequencing on the protective and nonprotective co-culture samples and identified substantial differences in the makeup of the bacterial communities. Protective bacterial communities consisted primarily of Alphaproteobacteria (Rhodobacteraceae) and Gammaproteobacteria (Marinobacter, Pseudomonas, Methylophaga) while nonprotective bacterial communities were less diverse and missing many putatively crucial members. We compared the seven protective communities with the seven nonprotective communities and we correlated specific bacterial amplicon sequence variants with algal protection. With these data, our future work will aim to define and develop an engineered-microbiome that can stabilize industrial Microchloropsis salina cultures by protecting against grazer-induced pond crashes.

Bioavailability of Mineral-Bound Iron to a Snow Algal-Bacterial Coculture and Implications for Albedo-Altering Snow Algal Blooms.

Snow algae can form large-scale blooms across the snowpack surface and near-surface environments. These pigmented blooms can decrease snow albedo and increase local melt rates, and they may impact the global heat budget and water cycle. Yet, the underlying causes for the geospatial occurrence of these blooms remain unconstrained. One possible factor contributing to snow algal blooms is the presence of mineral dust as a micronutrient source. We investigated the bioavailability of iron (Fe)-bearing minerals, including forsterite (Fo90, Mg1.8Fe0.2SiO4), goethite, smectite, and pyrite as Fe sources for a Chloromonas brevispina-bacterial coculture through laboratory-based experimentation. Fo90 was capable of stimulating snow algal growth and increased the algal growth rate in otherwise Fe-depleted cocultures. Fo90-bearing systems also exhibited a decrease in the ratio of bacteria to algae compared to those of Fe-depleted conditions, suggesting a shift in microbial community structure. The C. brevispina coculture also increased the rate of Fo90 dissolution relative to that of an abiotic control. Analysis of 16S rRNA genes in the coculture identified Gammaproteobacteria, Betaproteobacteria, and Sphingobacteria, all of which are commonly found in snow and ice environments. Archaea were not detected. Collimonas and Pseudomonas, which are known to enhance mineral weathering rates, comprised two of the top eight (>1%) operational taxonomic units (OTUs). These data provide unequivocal evidence that mineral dust can support elevated snow algal growth under otherwise Fe-depleted growth conditions and that snow algal microbial communities can enhance mineral dissolution under these conditions.IMPORTANCE Fe, a key micronutrient for photosynthetic growth, is necessary to support the formation of high-density snow algal blooms. The laboratory experiments described herein allow for a systematic investigation of the interactions of snow algae, bacteria, and minerals and their ability to mobilize and uptake mineral-bound Fe. Results provide unequivocal and comprehensive evidence that mineral-bound Fe in Fe-bearing Fo90 was bioavailable to Chloromonas brevispina snow algae within an algal-bacterial coculture. This evidence includes (i) an observed increase in snow algal density and growth rate, (ii) decreased ratios of bacteria to algae in Fo90-containing cultures relative to those of cultures grown under similarly Fe-depleted conditions with no mineral-bound Fe present, and (iii) increased Fo90 dissolution rates in the presence of algal-bacterial cocultures relative to those of abiotic mineral controls. These results have important implications for the role of mineral dust in supplying micronutrients to the snow microbiome, which may help support dense snow algal blooms capable of lowering snow albedo and increasing snow melt rates on regional, and possibly global, scales.

Microbial mutualism at a distance: The role of geometry in diffusive exchanges.

The exchange of diffusive metabolites is known to control the spatial patterns formed by microbial populations, as revealed by recent studies in the laboratory. However, the matrices used, such as agarose pads, lack the structured geometry of many natural microbial habitats, including in the soil or on the surfaces of plants or animals. Here we address the important question of how such geometry may control diffusive exchanges and microbial interaction. We model mathematically mutualistic interactions within a minimal unit of structure: two growing reservoirs linked by a diffusive channel through which metabolites are exchanged. The model is applied to study a synthetic mutualism, experimentally parametrized on a model algal-bacterial co-culture. Analytical and numerical solutions of the model predict conditions for the successful establishment of remote mutualisms, and how this depends, often counterintuitively, on diffusion geometry. We connect our findings to understanding complex behavior in synthetic and naturally occurring microbial communities.

Algal-bacterial cooperation improves algal photolysis-mediated hydrogen production

In this study, bacterium Pseudomonas sp. strain D was proved to be the main partner assisting Chlamydomonas reinhardtii in improving photolysis-mediated H2 production and a good partner for promoting H2 production by the green algae Chlorella and Scenedesmus. In strain D partnered algal-bacterial co-culture, the relative O2 content in the headspace plus the dissolved oxygen in the culture medium were rapidly consumed by bacterial growth, resulting in a completely anaerobic environment that proved suitable for the activation of algal hydrogenase. Moreover, algal-bacterial cooperation was able to slow the reduction of chlorophyll, enhance starch accumulation, and maintain protein content, which are the potential factors whose control provides an opportunity for improving algal H2 production. This study systematically analyzed the main pathway responsible for H2 production by algal-bacterial cooperation and the potential mechanisms for improvement, and proposed an efficient and durable algal-bacterial cooperation system for improving photolysis-mediated H2 production by green algae.

Factors influencing algal photobiohydrogen production in algal-bacterial co-cultures

Algal-bacterial co-cultures represent an alternative way for algal biohydrogen generation. Efficient algal hydrogen production requires anaerobiosis and electrons accessible for the algal FeFe‑hydrogenases. A number of factors strongly influence the development of this optimal environment. Various algal strains were tested for hydrogen evolution with a selected bacterial partner, a fully hydrogenase deficient Escherichia coli. During the hunt for the most efficient algae strains, gas-to-liquid phase ratio, algal optical density and algal cell size were identified as crucial factors influencing algal hydrogen evolution rate, accumulated algal hydrogen yield, carbon dioxide and oxygen levels as well as acetic acid consumption in illuminated algal-bacterial cultures. The highest accumulated hydrogen yields were observed for the different algal partners under similar experimental setup. The combination of a gas-to-liquid phase ratio of 1/1 with an algae cell density of 3.96∗108 algae cellml−1 (OD750: 1) resulted in the highest accumulated algal hydrogen yields under continuous illumination of ~50μmolm−2s−1 light at 25°C irrespective of the applied algae strain. Accumulated hydrogen yield was also strongly influenced by the algal cell size, smaller cell size correlated with higher hydrogen evolution rate. The highest accumulated algal hydrogen yield (88.98±2.19mlH2l−1d−1) was obtained with Chlorella sp. MACC 360 -E. coli ΔhypF co-culture.

Cryopreservation studies of an artificial co-culture between the cobalamin-requiring green alga Lobomonas rostrata and the bacterium Mesorhizobium loti

Algal-bacterial co-cultures, rather than cultures of algae alone, are regarded as having the potential to enhance productivity and stability in industrial algal cultivation. As with other inocula in biotechnology, to avoid loss of production strains, it is important to develop preservation methods for the long-term storage of these cultures, and one of the most commonly used approaches is cryopreservation. However, whilst there are many reports of cryopreserved xenic algal cultures, little work has been reported on the intentional preservation of both algae and beneficial bacteria in xenic cultures. Instead, studies have focused on the development of methods to conserve the algal strain(s) present, or to avoid overgrowth of bacteria in xenic isolates during the post-thaw recovery phase. Here, we have established a co-cryopreservation method for the long-term storage of both partners in a unialgal-bacterial co-culture. This is an artificial model mutualism between the alga Lobomonas rostrata and the bacterium Mesorhizobium loti, which provides vitamin B12 (cobalamin) to the alga in return for photosynthate. Using a Planer Kryo 360 controlled-rate cooler, post-thaw viability (PTV) values of 72% were obtained for the co-culture, compared to 91% for the axenic alga. The cultures were successfully revived after 6 months storage in liquid nitrogen, and continued to exhibit mutualism. Furthermore, the alga could be cryopreserved with non-symbiotic bacteria, without bacterial overgrowth occurring. It was also possible to use less controllable passive freezer chambers to cryopreserve the co-cultures, although the PTV was lower. Finally, we demonstrated that an optimised cryopreservation method may be used to prevent the overgrowth potential of non-symbiotic, adventitious bacteria in both axenic and co-cultures of L. rostrata after thawing.

Direct exchange of vitamin B12 is demonstrated by modelling the growth dynamics of algal–bacterial cocultures

The growth dynamics of populations of interacting species in the aquatic environment is of great importance, both for understanding natural ecosystems and in efforts to cultivate these organisms for industrial purposes. Here we consider a simple two-species system wherein the bacterium Mesorhizobium loti supplies vitamin B12 (cobalamin) to the freshwater green alga Lobomonas rostrata, which requires this organic micronutrient for growth. In return, the bacterium receives photosynthate from the alga. Mathematical models are developed that describe minimally the interdependence between the two organisms, and that fit the experimental observations of the consortium. These models enable us to distinguish between different mechanisms of nutrient exchange between the organisms, and provide strong evidence that, rather than undergoing simple lysis and release of nutrients into the medium, M. loti regulates the levels of cobalamin it produces, resulting in a true mutualism with L. rostrata. Over half of all microalgae are dependent on an exogenous source of cobalamin for growth, and this vitamin is synthesised only by bacteria; it is very likely that similar symbiotic interactions underpin algal productivity more generally.

실제 하수조건에서 조류-세균 복합군집의 생태적 상호작용 및 영양염류 제거 특성 규명

Abstract : Algal biomass cultivated by wastewater is potentially useful resource for biodiesel production. However, little is known about algal nutrient metabolism and microbial interaction with bacteria under real municipal wastewater condition. In this work, we characterized nitrogen and phosphorus removals of municipal wastewater by a representative wastewater-growing algal popu-lation. Ankistrodesmus gracilis SAG 278-2, and analyzed its ecological interaction with wastewater bacterial communities. Com-pared to wastewater sludge itself, algal-bacterial co-culture improved nutrient removal. According to bacterial community analysis with 16S rRNA genes, a selective and dominant growth of a Unclassified Alcaligenaceae population resulted from algal growth in the algal-bacterial co-culture. The selectively stimulated bacterial population is phylogenetically close to Alcaligenes faecalis subsp.5659-H, which is known to be co-present interact with algae in aquatic environment. These findings suggest that algal growth/ metabolism may have effects on selection of a specific bacterial population in algal-bacterial co-cultures that can efficiently removenutrients from municipal wastewater.