The 'Erlenmeter': a low-cost, open-source turbidimeter for no-sampling phenotyping of microorganism growth.

This work presents a low-cost, open-source turbidimeter, the 'Erlenmeter', designed to monitor the growth of microorganisms in batch cultures. It is easy to build, based exclusively on inexpensive off-the-shelf electronic components and 3D-printed parts. The Erlenmeter allows measuring the optical density of cultures on standard Erlenmeyer flasks without the need to open the flasks to collect aliquots, ensuring speed, minimal use of consumables, and elimination of the risk of contamination. These features make it particularly well-suited not just for routine research assays but also for experimental teaching. Here we illustrate the use of the Erlenmeter turbidimeter to record the growth of the microalga Phaeodactylum tricornutum, of the bacterium Escherichia coli, and of the yeast Saccharomyces cerevisiae, model organisms that are widely used in research and teaching. The Erlenmeter allows a detailed characterization of the growth curves of all organisms, confirming its usefulness for studying microbial populations dynamics both for research purposes and in classroom settings.

The Role of PGPB-Microalgae interaction in Alleviating Salt Stress in Plants.

Plant development and yield are severely hampered by climate change. Plants are very prone to a variety of abiotic stressors during growth, making them susceptible to destruction which can reduce the productivity by 20-60%. These stresses generate reactive oxygen species (ROS), which damage lipids, proteins, and nucleic acids. Microalgae and plant growth-promoting bacteria (PGPB) are remarkably effective at reducing the effects of salt stress and promoting plant growth, thereby increasing agricultural yield, and helping ensure global food security. Through a variety of mechanisms, including the production of phytohormones, 1-aminocyclopropane-1-carboxylic acid deaminase, exopolysaccharide, siderophores, hydrogen cyanide, extracellular polymeric substances, volatile organic compounds, and modulation of antioxidants defense machinery under abiotic stresses promote plant growth after inoculation of PGPB and microalgae. These microorganisms also maintain ion homeostasis, offer osmotic balance, stimulate genes that respond to salt and drought, rewire the metabolism, modify the transcription of ion transporter genes, and more. To counteract the negative consequences of salinity stress, this study summarizes the effects of PGPB- microalgae along with a tentative protective mechanism during salinity stress for sustainable agriculture.

High biomass yields of Chlorella protinosa with efficient nitrogen removal from secondary effluent in a membrane photobioreactor.

Further treatment of secondary effluents before their discharge into the receiving water bodies could alleviate water eutrophication. In this study, the Chlorella proteinosa was cultured in a membrane photobioreactor to further remove nitrogen from the secondary effluents. The effect of hydraulic retention time (HRT) on microalgae biomass yields and nutrient removal was studied. The results showed that soluble algal products concentration reduced in the suspension at low HRT, thereby alleviating microalgal growth inhibition. In addition, the lower HRT reduced the nitrogen limitation for Chlorella proteinosa's growth through the phase-out of nitrogen-related functional bacteria. As a result, the productivity for Chlorella proteinosa increased from 6.12 mg/L/day at an HRT of 24 hr to 20.18 mg/L/day at an HRT of 8 hr. The highest removal rates of 19.7 mg/L/day, 23.8 mg/L/day, and 105.4 mg/L/day were achieved at an HRT of 8 hr for total nitrogen (TN), ammonia, and chemical oxygen demand (COD), respectively. However, in terms of removal rate, TN and COD were the largest when HRT is 24 hr, which were 74.5% and 82.6% respectively. The maximum removal rate of ammonia nitrogen was 99.2% when HRT was 8 hr.

Influence of chitosan properties and operating parameters on the flocculation efficiency and harvesting of microalgae (Scenedesmus sp.).

Physicochemical and structural characteristics of chitosan prepared from Deep-sea shrimp (DCs), including degree of deacetylation (DD), molecular weight (Mw), viscosity, crystallinity index (CrI) and surface morphology were compared with a commercial chitosan (CCs). The DCs had a higher DD of 81.33 ± 0.40 %, whereas the CCs had a lower DD of 74.62 ± 0.64 %. Additionally, the DCs exhibited a lower Mw of 192.47 ± 2.5 kDa and viscosity of 646.00 ± 4.00 cP compared to the CCs, which had a Mw of 202.44 ± 0.28 kDa and viscosity of 689.67 ± 5.91 cP. This study investigated the influence of chitosan properties, particularly DD and Mw on the harvesting of Scenedesmus sp. along with the chitosan dosage, pH of the culture medium, mixing speed and time. Under optimal operating conditions, the microalgae removal efficiency of the DCs reached a significantly higher level (94.71 ± 0.20 %) compared to that of CCs (88.25 ± 0.41 %). Chitosan with a higher DD and low Mw demonstrated superior flocculation efficiency. The results highlight the significance of DD and Mw of chitosan and its influence on the flocculation of microalgae, providing valuable insights for optimizing the harvesting process with the non-toxic and natural flocculent, chitosan.

Metabolomics Reveals the Impact of Overexpression of Cytosolic Fructose-1,6-Bisphosphatase on Photosynthesis and Growth in Nannochloropsis gaditana.

Nannochloropsis gaditana, a microalga known for its photosynthetic efficiency, serves as a cell factory, producing valuable biomolecules such as proteins, lipids, and pigments. These components make it an ideal candidate for biofuel production and pharmaceutical applications. In this study, we genetically engineered N. gaditana to overexpress the enzyme fructose-1,6-bisphosphatase (cyFBPase) using the Hsp promoter, aiming to enhance sugar metabolism and biomass accumulation. The modified algal strain, termed NgFBP, exhibited a 1.34-fold increase in cyFBPase activity under photoautotrophic conditions. This modification led to a doubling of biomass production and an increase in eicosapentaenoic acid (EPA) content in fatty acids to 20.78-23.08%. Additionally, the genetic alteration activated the pathways related to glycine, protoporphyrin, thioglucosides, pantothenic acid, CoA, and glycerophospholipids. This shift in carbon allocation towards chloroplast development significantly enhanced photosynthesis and growth. The outcomes of this study not only improve our understanding of photosynthesis and carbon allocation in N. gaditana but also suggest new biotechnological methods to optimize biomass yield and compound production in microalgae.

General performance, kinetic modification, and key regulating factor recognition of microalgae-based sulfonamide removal.

Sulfonamides have been widely detected in water treatment plants. Advanced wastewater treatment for sulfonamide removal based on microalgal cultivation can reduce the ecological risk after discharge, achieve carbon fixation, and simultaneously recover bioresource. However, the general removal performance, key factors and their impacts, degradation kinetics, and potential coupling technologies have not been systematically summarized. To guide the construction and enhance the efficient performance of the purification system, this study summarizes the quantified characteristics of sulfonamide removal based on more than 100 groups of data from the literature. The biodegradation potential of sulfonamides from different subclasses and their toxicity to microalgae were statistically analyzed; therefore, a preferred option for further application was proposed. The mechanisms by which the properties of both sulfonamides and microalgae affect sulfonamide removal were comprehensively summarized. Thereafter, multiple principles for choosing optimal microalgae were proposed from the perspective of engineering applications. Considering the microalgal density and growth status, a modified antibiotic removal kinetic model was proposed with significant physical meaning, thereby resulting in an optimal fit. Based on the mechanism and regulating effect of key factors on sulfonamide removal, sensitive and feasible factors (e.g., water quality regulation, other than initial algal density) and system coupling were screened to guide engineering applications. Finally, we suggested studying the long-term removal performance of antibiotics at environmentally relevant concentrations and toxicity interactions for further research.

The impact of polystyrene nanoplastics (PSNPs) on physiological and biochemical parameters of the microalgae Spirulina platensis.

Nanoplastics, as emerging pollutants, have harmful effects on living organisms and the environment, the mechanisms and extent of which remain unclear. Microalgae, as one of the most important biological groups in the food chain and sensitive environmental indicators to various pollutants, are considered a suitable option for investigating the effects of nanoplastics. In this study, the effects of polystyrene nanoplastics on the growth rate, dry weight, chlorophyll a and carotenoid levels, proline, and lipid peroxidation in the Spirulina platensis were examined. Three concentrations of 0.1, 1, and 10 mg L-1 of PSNPs were used alongside a control sample with zero concentration, with four repetitions in one-liter containers for 20 days under optimal temperature and light conditions. Various analyses, including growth rate, dry weight, proline, chlorophyll a and carotenoid levels, and lipid peroxidation, were performed. The results indicated that exposure to PSNP stress led to a significant decrease in growth rate, dry weight, and chlorophyll a and carotenoid levels compared to the control sample. Furthermore, this stress increased the levels of proline and lipid peroxidation in Spirulina platensis. Morphological analysis via microscopy supported these findings, indicating considerable environmental risks associated with PSNPs.

Performance evaluation of typical flocculants for efficient harvesting of Chlorella sorokiniana under different carbon application modes.

In this study, the growth characteristics of microalgae cultured with different carbon sources were analyzed, and the flocculation characteristics under the influence of carbon sources were evaluated using three typical flocculants. The results showed that the organic carbon sources could significantly increase the content of extracellular proteins in microalgae. Specifically, the extracellular protein concentrations of microalgae cultured with pure BG-11, ethanol, sodium acetate and glucose were 18.2 29.2, 97.3, and 34.7 mg/g, respectively. During the flocculation process, microalgae cultured with sodium acetate exhibited a weak response to the flocculant because of excessive extracellular proteins inhibited flocculation. In addition, the flocculation efficiency was also less than 50.0% cultured with sodium acetate in all pH test ranges when alum and chitosan were used as flocculants. It could be inferred that the flocculant initially happened to charge neutralization with the negatively charged proteins in the solution and then bridged the charges with the microalgae. These findings provide insights into the effects of different carbon sources on microalgal flocculation, promising organic integration of microalgae wastewater treatment and harvesting.

Enhancing environmental performance in biogas production from wastewater-grown microalgae: A life cycle assessment perspective.

The production of biogas from microalgae has gained attention due to their rapid growth, CO2 sequestration, and minimal land use. This study uses life cycle assessment to assess the environmental impacts of biogas production from wastewater-grown microalgae through anaerobic digestion within an optimized microalgae-based system. Using SimaPro® 9 software, 3 scenarios were modeled considering the ReCiPe v1.13 midpoint and endpoint methods for environmental impact assessment in different categories. In the baseline scenario (S1), a hypothetical system for biogas production was considered, consisting of a high rate algal pond (HRAP), a settling, an anaerobic digester, and a biogas upgrading unit. The second scenario (S2) included strategies to enhance biogas yield, namely co-digestion and thermal pre-treatment. The third scenario (S3), besides considering the strategies of S2, proposed the biogas upgrading in the HRAP and the digestate recovery as a biofertilizer. After normalization, human carcinogenic toxicity was the most positively affected category due to water use in the cultivation step, accounted as avoided product. However, this category was also the most negatively affected by the impacts of the digester heating energy. Anaerobic digestion was the most impactful step, constituting on average 60.37% of total impacts. Scenario S3 performed better environmentally, primarily due to the integration of biogas upgrading within the cultivation reactor and digestate use as a biofertilizer. Sensitivity analysis highlighted methane yield's importance, showing potential for an 11.28% reduction in ionizing radiation impacts with a 10% increase. Comparing S3 biogas with natural gas, the resource scarcity impact was reduced sixfold, but the human health impact was 23 times higher in S3.

Effect of microalgae and bacteria inoculation on the startup of bioreactors for paper pulp wastewater and biofuel production.

The use of microalgae and bacteria as a strategy for the startup of bioreactors for the treatment of industrial wastewater can be a sustainable and economically viable alternative. This technology model provides satisfactory results in the nitrification and denitrification process for nitrogen removal, organic matter removal, biomass growth, sedimentation, and byproducts recovery for added-value product production. The objective of this work was to evaluate the performance of microalgae and bacteria in their symbiotic process when used in the treatment of paper pulp industry wastewater. The experiment, lasting fourteen days, utilized four bioreactors with varying concentrations in mgVSS/L of microalgae to bacteria ratio (R1-100:100, R2-100:300, R3-100:500, R4-300:100) in the startup process. Regarding the sludge volumetric index (SVI), the results show that the R1 and R2 reactors developed SVI30/SVI10 biomass in the range of 85.57 ± 7.33% and 84.72 ± 8.19%, respectively. The lipid content in the biomass of reactors R1, R2, R3 e R4 was 13%, 7%, 19%, and 22%, respectively. This high oil content at the end of the batch, may be related to the nutritional stress that the species underwent during this feeding regime. In terms of chlorophyll, the bioreactor with an initial inoculation of 100:100 showed better symbiotic growth of microalgae and bacteria, allowing exponential growth of microalgae. The total chlorophyll value for this bioreactor was 801.46 ± 196.96 µg/L. Biological removal of nitrogen from wastewater from the paper pulp industry is a challenge due to the characteristics of the effluent, but the four reactors operated in a single batch obtained good nitrogen removal. Ammonia nitrogen removal performances were 91.55 ± 9.99%, 72.13 ± 19.18%, 64.04 ± 21.34%, and 86.15 ± 30.10% in R1, R2, R3, and R4, respectively.

Cultivation of yeasts on liquid digestate to remove organic pollutants and nutrients and for potential application as co-culture with microalgae.

In this study, the growth of yeast and yeast-like fungi in the liquid digestate from vegetable wastes was investigated in order to remove nutrients and organic pollutants, and for their application as co-culture members with green microalgae. The studied yeast strains were characterized for their assimilative and enzymatic profiles as well as temperature requirements. In the first experimental stage, the growth dynamics of each strain were determined, allowing to select the best yeasts for further studies. In the subsequent stage, the ability of selectants to remove organic pollutants was assessed. Different cultivation media containing respectively 1:3, 1:1, 3:1 vol ratio of liquid digestate and the basal minimal medium were used. Among all tested yeast strains, Rhodotorula mucilaginosa DSM 70825 showed the most promising results, demonstrating the highest potential for removing organic substrates and nutrients. Depending on the medium, this strain achieved 50-80% sCOD, 45-60% tVFAs, 21-45% TN, 33-52% PO43- reduction rates. Similar results were obtained for the strain Candida sp. OR687571. The high nutrient and organics removal efficiency by these yeasts could likely be linked to their ability to assimilate xylose (being the main source of carbon in the liquid digestate). In culture media containing liquid digestate, both yeast strains achieved good viability and proliferation potential. In the liquid digestate medium, R. mucilaginosa and Candida sp. showed vitality at the level of 51.5% and 45.0%, respectively. These strains seem to be a good starting material for developing effective digestate treatment strategies involving monocultures and/or consortia with other yeasts or green microalgae.

Central transcriptional regulator controls photosynthetic growth and carbon storage in response to high light.

Carbon capture and biochemical storage are some of the primary drivers of photosynthetic yield and productivity. To elucidate the mechanisms governing carbon allocation, we designed a photosynthetic light response test system for genetic and metabolic carbon assimilation tracking, using microalgae as simplified plant models. The systems biology mapping of high light-responsive photophysiology and carbon utilization dynamics between two variants of the same Picochlorum celeri species, TG1 and TG2 elucidated metabolic bottlenecks and transport rates of intermediates using instationary 13C-fluxomics. Simultaneous global gene expression dynamics showed 73% of the annotated genes responding within one hour, elucidating a singular, diel-responsive transcription factor, closely related to the CCA1/LHY clock genes in plants, with significantly altered expression in TG2. Transgenic P. celeri TG1 cells expressing the TG2 CCA1/LHY gene, showed 15% increase in growth rates and 25% increase in storage carbohydrate content, supporting a coordinating regulatory function for a single transcription factor.

Microalgae cultivation trials in a membrane bioreactor operated in heterotrophic, mixotrophic, and phototrophic modes using ammonium-rich wastewater: The study of fouling.

In this work, microalgae cultivation trials were carried out in a membrane bioreactor to investigate fouling when the cultures of Chlorellavulgaris were grown under mixotrophic, heterotrophic, and phototrophic cultivation regimes. The Chlorella cultures were cultivated in wastewater as a source of nutrients that contained a high concentration of ammonium. In mixotrophic cultivation trials, the results showed that the elevated contents of carbohydrates in the soluble microbial product and proteins in extracellular polymeric substances probably initiated membrane fouling. In this case, the highest protein content was also found in extracellular polymeric substances due to the high nitrogen removal rate. Consequently, transmembrane pressure significantly increased compared to the phototrophic and heterotrophic regimes. The data indicated that cake resistance was the main cause of fouling in all cultivations. Higher protein content in the cake layer made the membrane surface more hydrophobic, while carbohydrates had the opposite effect. Compared to a mixotrophic culture, a phototrophic culture had a larger cell size and higher hydrophobicity, leading to less membrane fouling. Based on our previous data, the highest ammonia removal rate was reached in the mixotrophic cultures; nevertheless, membrane fouling appeared to be the fundamental problem.

Physical structure of the environment contributes to the development of diversity of microalgal assemblages.

Aquatic macrophytes form a three dimensional complex structure in the littoral zones of lakes, with many physical, chemical and biological gradients and interactions. This special habitat harbours a unique microalgal assemblage called metaphyton, that differs both from the phytoplankton of the pelagial and from the benthic assemblages whose elements are tightly attached to the substrates. Since metaphytic assemblages significantly contribute to the diversity of lakes' phytoplankton, it is crucial to understand and disentangle those mechanisms that ensure their development. Therefore, we focused on the question of how a single solid physical structure contribute to maintaining metaphytic assemblages. Using a laboratory experiment we studied the floristic and functional differences of microalgal assemblages in microcosms that simulated the conditions that an open water, a complex natural macrophyte stand (Utricularia vulgaris L.), or an artificial substrate (cotton wool) provide for them. We inoculated the systems with a species rich (> 326 species) microalgal assemblage collected from a eutrophic oxbow lake, and studied the diversity, trait and functional group composition of the assemblages in a 24 day long experimental period. We found that both natural and artificial substrates ensured higher species richness than the open water environment. Functional richness in the open water environment was lower than in the aquaria containing natural macrophyte stand but higher than in which cotton wool was placed. This means that the artificial physical structure enhanced functional redundancy of the resident functional groups. Elongation measures of microalgal assemblages showed the highest variation in the microcosms that simulated the open water environment. Our results suggest that assembly of metaphytic algal communities is not a random process, instead a deterministic one driven by the niche characteristics of the complex three dimensional structure created by the stands of aquatic macrophytes.

The potential of Pavlovophyceae species as a source of valuable carotenoids and polyunsaturated fatty acids for human consumption.

Microalgae are a group of microorganisms, mostly photoautotrophs with high CO2 fixation capacity, that have gained increased attention in the last decades due to their ability to produce a wide range of valuable metabolites, such as carotenoids and polyunsaturated fatty acids, for application in food/feed, pharmaceutical, and cosmeceutical industries. Their increasing relevance has highlighted the importance of identifying and culturing new bioactive-rich microalgae species, as well as of a thorough understanding of the growth conditions to optimize the biomass production and master the biochemical composition according to the desired application. Thus, this review intends to describe the main cell processes behind the production of carotenoids and polyunsaturated fatty acids, in order to understand the possible main triggers responsible for the accumulation of those biocompounds. Their economic value and the biological relevance for human consumption are also summarized. In addition, an extensive review of the impact of culture conditions on microalgae growth performance and their biochemical composition is presented, focusing mainly on the studies involving Pavlovophyceae species. A complementary description of the biochemical composition of these microalgae is also presented, highlighting their potential applications as a promising bioresource of compounds for large-scale production and human and animal consumption.

Application of hybrid multi-criteria decision-making approach to analyze wastewater microalgae culture systems for bioenergy production.

Bioenergy generation from microalgae can significantly contribute to climate mitigation and renewable energy production. In this regard, several multi-criteria decision-making method were employed to prioritize appropriate microalgae culture system for bioenergy production. Entropy weight, Criteria Importance Through Intercriteria Correlation (CRITIC) and Technique for Order of Preference by Similarity to Ideal Solution (TOPSIS) were the employed MCDA method. Fourteen microalgae culture systems were selected as a case study, which contain teen monoculture and four dual-culture. Initially, through ans in-depth review of the literature and expert views, four categories total eight indicators were selected as the evaluation indices of the study, namely 1) Proliferation: Half growth cycle and Max growth rate,2) Biomass output: Bio-crude yield and Lipid yield, 3) Nutrient utilization: residual concentration of total Nitrogen and total Phosphorus, and, 4) Stability: coefficient of variation of Bio-crude yield and Lipid yield. The result indicated that "Pediastrum sp. & Micractinium sp." was identified as the most bioenergy potential microalgae culture system, and the evaluation results of entropy weight method and CRITIC method are similar. It is pertinent to note that 1)the entropy weight method exhibits lower sample size requirements, 2) the critic method excels when dealing with larger sample sizes, and 3) the TOPSIS method necessitates the incorporation of appropriate weighting methods to ensure credible results. In the application stage, the key indicators related to cost can be further included in the evaluation indices.

Light management by algal aggregates in living photosynthetic hydrogels.

Rapid progress in algal biotechnology has triggered a growing interest in hydrogel-encapsulated microalgal cultivation, especially for the engineering of functional photosynthetic materials and biomass production. An overlooked characteristic of gel-encapsulated cultures is the emergence of cell aggregates, which are the result of the mechanical confinement of the cells. Such aggregates have a dramatic effect on the light management of gel-encapsulated photobioreactors and hence strongly affect the photosynthetic outcome. To evaluate such an effect, we experimentally studied the optical response of hydrogels containing algal aggregates and developed optical simulations to study the resultant light intensity profiles. The simulations are validated experimentally via transmittance measurements using an integrating sphere and aggregate volume analysis with confocal microscopy. Specifically, the heterogeneous distribution of cell aggregates in a hydrogel matrix can increase light penetration while alleviating photoinhibition more effectively than in a flat biofilm. Finally, we demonstrate that light harvesting efficiency can be further enhanced with the introduction of scattering particles within the hydrogel matrix, leading to a fourfold increase in biomass growth. Our study, therefore, highlights a strategy for the design of spatially efficient photosynthetic living materials that have important implications for the engineering of future algal cultivation systems.

Inorganic salt starvation improves the polysaccharide production and CO2 fixation by Porphyridium purpureum.

The microalgae industry shows a promising future in the production of high-value products such as pigments, phycoerythrin, polyunsaturated fatty acids, and polysaccharides. It was found that polysaccharides have high biomedical value (such as antiviral, antibacterial, antitumor, antioxidative) and industrial application prospects (such as antioxidants). This study aimed to improve the polysaccharides accumulation of Porphyridium purpureum CoE1, which was effectuated by inorganic salt starvation strategy whilst supplying rich carbon dioxide. At a culturing temperature of 25 °C, the highest polysaccharide content (2.89 g/L) was achieved in 50% artificial seawater on the 12th day. This accounted for approximately 37.29% of the dry biomass, signifying a 25.3% increase in polysaccharide production compared to the culture in 100% artificial seawater. Subsequently, separation, purification and characterization of polysaccharides produced were conducted. Furthermore, the assessment of CO2 fixation capacity during the cultivation of P. purpureum CoE1 was conducted in a 10 L photobioreactor. This indicated that the strain exhibited an excellent CO2 fixation capacity of 1.66 g CO2/g biomass/d. This study proposed an efficient and feasible approach that not only increasing the yield of polysaccharides by P. purpureum CoE1, but also fixing CO2 with a high rate, which showed great potential in the microalgae industry and Bio-Energy with Carbon Capture and Storage.

Abreast insights of harnessing microalgal lipids for producing biodiesel: A review of improving and advancing the technical aspects of cultivation.

In the pursuit of alternatives for conventional diesel, sourced from non-renewable fossil fuel, biodiesel has gained attentions for its intrinsic benefits. However, the commercial production process for biodiesel is still not sufficiently competitive. This review analyses microalgal lipid, one of the important sources of biodiesel, and its cultivation techniques with recent developments in the technical aspects. In fact, the microalgal lipids are the third generation feedstock, used for biodiesel production after its benefits outweigh that of edible vegetable oils (first generation) and non-edible oils (second generation). The critical factors influencing microalgal growth and its lipid production and accumulation are also discussed. Following that is the internal enhancement for cellular lipid production through genetic engineering. Moreover, the microalgae cultivation data modelling was also rationalized, with a specific focus on growth kinetic models that allow for the prediction and optimization of lipid production. Finally, the machine learning and environmental impact analysis are as well presented as important aspects to consider in fulfilling the prime objective of commercial sustainability to produce microalgal biodiesel.

Temperature-controlled microalgal biofilm detachment and harvesting assisted by ultrasonic from 3D porous substrates grafted with thermosensitive gels.

Microalgae have been renowned as the most promising energy organism with significant potential in carbon fixation. In the large-scale cultivation of microalgae, the 3D porous substrate with higher specific surface area is favorable to microalgae adsorption and biofilm formation, whereas difficult for biofilm detachment and microalgae harvesting. To solve this contradiction, N-isopropylacrylamide, a temperature-responsive gels material, was grafted onto the inner surface of the 3D porous substrate to form temperature-controllable interface wettability. The interfacial free energy between microalgae biofilm and the substrates increased from -63.02 mJ/m2 to -31.89 mJ/m2 when temperature was lowered from 32 °C to 17 °C, weakening the adsorption capacity of cells to the surface, and making the biofilm detachment ratio increased to 50.8%. When further cooling the environmental temperature to 4 °C, the detachment capability of microalgae biofilm kept growing. 91.6% of the cells in the biofilm were harvesting from the 3D porous substrate. And the biofilm detached rate was up to 19.84 g/m2/h, realizing the temperature-controlled microalgae biofilm harvesting. But, microalgae growth results in the secretion of extracellular polymeric substances (EPS), which enhanced biofilm adhesion and made cell detachment more difficult. Thus, ultrasonic vibration was used to reinforce biofilm detachment. With the help of ultrasonic vibration, microalgae biofilm detached rate increased by 143.45% to 41.07 g/m2/h. These findings provide a solid foundation for further development of microalgae biofilm detachment and harvesting technology.