Marine Protists Research Articles

Enhancing fatty acid and omega-3 production in Schizochytrium sp. using developed safe-harboring expression system

BackgroundSchizochytrium, a group of eukaryotic marine protists, is an oleaginous strain, making it a highly promising candidate for the production of lipid-derived products such as biofuels and omega-3 fatty acids. However, the insufficient advancement of genetic engineering tools has hindered further advancements. Therefore, the development and application of genetic engineering tools for lipid enhancement are crucial for industrial production.ResultsTransgene expression in Schizochytrium often encounters challenges such as instability due to positional effects. To overcome this, we developed a safe-harbor transgene expression system. Initially, the sfGFP gene was integrated randomly, and high-expressing transformants were identified using fluorescence-activated cell sorting. Notably, HRsite 2, located approximately 3.2 kb upstream of cytochrome c, demonstrated enhanced sfGFP expression and homologous recombination efficiency. We then introduced the 3-ketoacyl-ACP reductase (KR) gene at HRsite 2, resulting in improved lipid and docosahexaenoic acid (DHA) production. Transformants with KR at HRsite 2 exhibited stable growth, increased glucose utilization, and a higher lipid content compared to those with randomly integrated transgenes. Notably, these transformants showed a 25% increase in DHA content compared to the wild-type strain.ConclusionThis study successfully established a robust homologous recombination system in Schizochytrium sp. by identifying a reliable safe harbor site for gene integration. The targeted expression of the KR gene at this site not only enhanced DHA production but also maintained growth and glucose consumption rates, validating the efficacy of the safe-harbor approach. This advancement in synthetic biology and metabolic engineering paves the way for more efficient biotechnological applications in Schizochytrium sp.

How marine are Marine Stramenopiles (MAST)? A cross-system evaluation.

Marine Stramenopiles (MAST) were first described two decades ago through ribosomal RNA gene (rRNA gene) sequences from marine surveys of microbial eukaryotes. MAST comprise several independent lineages at the base of the Stramenopiles. Despite their prevalence in the ocean, the majority of MAST diversity remains uncultured. Previous studies, mainly in marine environments, have explored MAST's cell morphology, distribution, trophic strategies, and genomics using culturing-independent methods. In comparison, less is known about their presence outside marine habitats. Here, we analyze the extensive EukBank dataset to assess the extent to which MAST can be considered marine protists. Additionally, by incorporating newly available rRNA gene sequences, we update Stramenopiles phylogeny, identifying three novel MAST lineages. Our results indicate that MAST are primarily marine with notable exceptions within MAST-2 and MAST-12, where certain subclades are prevalent in freshwater and soil habitats. In the marine water column, only a few MAST species, particularly within clades -1, -3, -4 and -7, dominate and exhibit clear latitudinal distribution patterns. Overall, the massive sequencing dataset analyzed in our study confirms and partially expands the previously described diversity of MASTs groups and underscores the predominantly marine nature of most of these uncultured lineages.

Aurantiochytrium mutant strains exhibiting different colony colors altered the contents of squalene.

Aurantiochytrium sp. 18W-13a, a marine heterotrophic protist belonging to the genus thraustochytrid, is known to accumulate high levels of squalene and carotenoids. Nowadays, the mutagenesis breeding of microorganisms is still widely practiced because the induced mutations of DNA do not involve the permanent integration of heterologous DNA sequences. Therefore, in this study, we focused on the improvement of squalene yield by mutagenesis breeding using Aurantiochytrium sp. 18W-13a. To bypass the massively laborious screening, we propose to use colony colors as the first criterion to screen mutants with high squalene accumulation, since the carotenoid and squalene synthetic pathways share an intermediate. We selected pale (white)-colored mutants after carbon ion irradiation. The white mutants exhibited larger squalene yields than twice as much of the original strain. The results clearly indicate that the present screening method with colony colors promises to obtain productive strains of squalene.

Global distribution, diversity, and ecological niche of Picozoa, a widespread and enigmatic marine protist lineage

BackgroundThe backbone of the eukaryotic tree of life contains taxa only found in molecular surveys, of which we still have a limited understanding. Such is the case of Picozoa, an enigmatic lineage of heterotrophic picoeukaryotes within the supergroup Archaeplastida, which has emerged as a significant component of marine microbial planktonic communities. To enhance our understanding of the diversity, distribution, and ecology of Picozoa, we conduct a comprehensive assessment at different levels, from assemblages to taxa, employing phylogenetic analysis, species distribution modeling, and ecological niche characterization.ResultsPicozoa was among the ten most abundant eukaryotic groups, found almost exclusively in marine environments. The phylum was represented by 179 Picozoa’s OTU (pOTUs) placed in five phylogenetic clades. Picozoa community structure had a clear latitudinal pattern, with polar assemblages tending to cluster separately from non-polar ones. Based on the abundance and occupancy pattern, the pOTUs were classified into four categories: Low-abundant, Widespread, Polar, and Non-polar. We calculated the ecological niche of each of these categories. Notably, pOTUs sharing similar ecological niches were not closely related species, indicating a phylogenetic overdispersion in Picozoa communities. This could be attributed to competitive exclusion and the strong influence of the seasonal amplitude of variations in environmental factors, such as temperature, shaping physiological and ecological traits.ConclusionsOverall, this work advances our understanding of uncharted protists’ evolutionary dynamics and ecological strategies. Our results highlight the importance of understanding the species-level ecology of marine heteroflagellates like Picozoa. The observed phylogenetic overdispersion challenges the concept of phylogenetic niche conservatism in protist communities, suggesting that closely related species do not necessarily share similar ecological niches.9DjsWS8J_yUMTxizMfQ8tyVideo

Phylogenetic proximity drives temporal succession of marine giant viruses in a five-year metagenomic time-series.

Nucleocytoplasmic Large DNA Viruses (NCLDVs, also called giant viruses) are widespread in marine systems and infect a broad range of microbial eukaryotes (protists). Recent biogeographic work has provided global snapshots of NCLDV diversity and community composition across the world's oceans, yet little information exists about the guiding 'rules' underpinning their community dynamics over time. We leveraged a five-year monthly metagenomic time-series to quantify the community composition of NCLDVs off the coast of Southern California and characterize these populations' temporal dynamics. NCLDVs were dominated by Algavirales (Phycodnaviruses, 59%) and Imitervirales (Mimiviruses, 36%). We identified clusters of NCLDVs with distinct classes of seasonal and non-seasonal temporal dynamics. Overall, NCLDV population abundances were often highly dynamic with a strong seasonal signal. The Imitervirales group had highest relative abundance in the more oligotrophic late summer and fall, while Algavirales did so in winter. Generally, closely related strains had similar temporal dynamics, suggesting that evolutionary history is a key driver of the temporal niche of marine NCLDVs. However, a few closely-related strains had drastically different seasonal dynamics, suggesting that while phylogenetic proximity often indicates ecological similarity, occasionally phenology can shift rapidly, possibly due to host-switching. Finally, we identified distinct functional content and possible host interactions of two major NCLDV orders-including connections of Imitervirales with primary producers like the diatom Chaetoceros and widespread marine grazers like Paraphysomonas and Spirotrichea ciliates. Together, our results reveal key insights on season-specific effect of phylogenetically distinct giant virus communities on marine protist metabolism, biogeochemical fluxes and carbon cycling.

Rhizaria in the oligotrophic ocean exhibit clear temporal and vertical variability

Recently studies have shown that Rhizaria, a super-group of marine protists, have a large role in pelagic ecosystems. They are unique in that they construct mineral tests out of silica, calcium carbonate, or strontium sulfate. As a consequence, Rhizaria can have large impacts on the ocean’s cycling of carbon and other elements. However, less is known about Rhizaria ecology or their role in the pelagic food-web. Some taxa, like certain Radiolarians, are mixotrophic, hosting algal symbionts. While other taxa are flux-feeders or even predatory carnivores. Some prior research has suggested that Rhizaria will partition vertically in the water column, likely due to different trophic strategies. However, very few studies have investigated their populations over extended periods of time. In this study, we present data investigating Rhizaria abundance and vertical distribution from over a year of monthly cruises in the Sargasso Sea. This study represents the first quantification of Rhizaria throughout the mesopelagic zone in an oligotrophic system for an extended period of time. We use this data to investigate the hypothesis that Rhizaria taxonomic groups will partition due to trophic mode. We also investigate how their abundance varies in accordance with environmental parameters. Rhizaria abundance was quantified using an Underwater Vision Profiler (UVP5), an in-situ imaging device. Ultimately, we show that different Rhizaria taxa will have unique vertical distribution patterns. Models relating their abundance to environmental parameters have mixed results, yet particle concentration is a common predictive variable, supporting the importance of heterotrophy amongst many taxa.

High resolution 3D images of sediment cores as powerful tool for exploring foraminiferal microhabitats

Benthic foraminifera are marine protists largely used as bioindicators and proxies of paleo- environments. Epifaunal species are supposed to live at or above the sediment surface and are therefore used as proxies for bottom water conditions, while infaunal inhabit the sediment column, thus tracing porewater chemistry. Traditional analytical methods based on core slicing, however, have a low resolution that does not allow to precisely characterise the preferential microhabitat(s) of indicator species.In this study we performed microtomographic analyses on an experimental sediment core, to observe the life-position of living foraminifera of two surface-dwelling species Ammonia confertitesta and Haynesina germanica, reported both as epifaunal or shallow infaunal. The images we obtained offered for the first time the possibility to observe each individual in 3D space with a numerical resolution of 13 μm/voxel.The results revealed that the two species are never located above or at the sediment surface and have their preferential microhabitats in a sub-superficial sediment layer constrained in the 0–500 μm interval below the surface. Rapid decrease of abundances below this layer suggests that their microhabitat could be even more specific than previously thought.μCT-scan of sediment cores is also a valuable tool to obtain high-resolution information about foraminiferal ecology. The described method is useful to assess the effective microhabitat of all foraminiferal species that are usually used as proxies for paleorecords, to ensure that the information we can obtain from them is attributable to bottom water or to porewater conditions at a specific sediment depth.

Transient, context-dependent fitness costs accompanying viral resistance in isolates of the marine microalga Micromonas sp. (class Mamiellophyceae).

Marine microbes are important in biogeochemical cycling, but the nature and magnitude of their contributions are influenced by their associated viruses. In the presence of a lytic virus, cells that have evolved resistance to infection have an obvious fitness advantage over relatives that remain susceptible. However, susceptible cells remain extant in the wild, implying that the evolution of a fitness advantage in one dimension (virus resistance) must be accompanied by a fitness cost in another dimension. Identifying costs of resistance is challenging because fitness is context-dependent. We examined the context dependence of fitness costs in isolates of the picophytoplankton genus Micromonas and their co-occurring dsDNA viruses using experimental evolution. After generating 88 resistant lineages from two ancestral Micromonas strains, each challenged with one of four distinct viral strains, we found resistance led to a 46% decrease in mean growth rate under high irradiance and a 19% decrease under low. After a year in culture, the experimentally selected lines remained resistant, but fitness costs had attenuated. Our results suggest that the cost of resistance in Micromonas is dependent on environmental conditions and the duration of population adaptation, illustrating the dynamic nature of fitness costs of viral resistance among marine protists.

Assessing the performance of short 18S rDNA markers for environmental DNA metabarcoding of marine protists

AbstractMarine protists are globally distributed and sensitive to environmental conditions, which makes them a focal group when studying the effects of climate change on biodiversity and ocean health. However, they are a highly diverse group with varying evolutionary histories and morphologies and widely variable preservation potential in the fossil record. Thus, their past diversity and composition are poorly known. Paleogenetics, which relies, among other approaches, on DNA metabarcoding of sedimentary ancient DNA (sedaDNA), provides a promising avenue to explore the past history and responses of marine protists to global change. Choosing the right marker for sedaDNA studies is critical, striking a balance between marker length and taxonomic resolution. While marker guides exist for modern environmental DNA surveys, a thorough assessment of existing short markers for sedaDNA studies targeting protists is lacking. In this study, we report on a comparison of in silico PCR for eight short 18S rDNA markers, including one from the Tara Oceans initiative and a longer marker commonly used in modern marine eDNA studies. We analyze their taxonomic coverage and resolution, taxonomic overlap and uniqueness between markers, co‐amplification of non‐protist taxa, and amplicon size differences across taxonomic groups. Additionally, we provide a detailed analysis of diatoms, dinoflagellates, haptophytes, and chlorophytes. Our study is aimed at supporting project‐specific marker choices for characterizing protist composition and diversity. While we focus on marine protists, our results are applicable to other aquatic and terrestrial environments.

Surface currents shape protist community structure across the Indo-Pacific.

Biogeographic structure in marine protist communities is shaped by a combination of dispersal potential and environmental selection. High-throughput sequencing and global sampling efforts have helped better resolve the composition and functions of these communities in the world's oceans using both molecular and visual methods. However, molecular barcoding data are critically lacking across the Indo-Pacific, a region widely considered the epicenter of marine biodiversity. To fill this gap, we characterized protist communities in four sampling regions across Indonesia that represent the latitudinal, longitudinal, and human population gradients of the region: Lombok, Wakatobi, Misool, and Waigeo. We show high spatial structuring in marine protist communities across Indonesia, and biotic factors appear to play little role in driving this observed structure. Our results appear to be driven by abiotic factors linked to surface current patterns across the Indo-Pacific as a result of: (1) a choke point in circulation at the Indonesian Throughflow leading to low diatom diversity in Lombok, Wakatobi, and Misool; (2) an increase in nutrient availability at the edge of the Halmahera Eddy in Waigeo, leading to an increase in diatom diversity; and/or (3) seasonal variations in protist communities in line with shifts in velocity of the Indonesian Throughflow. Overall, our results highlight the importance of abiotic factors in shaping protist communities on broad geographic scales over biotic, top-down pressures, such as grazing from higher trophic levels.

Optimal filter materials for protist quantification via droplet digital PCR

The use of droplet digital polymerase chain reaction (ddPCR) has greatly improved the quantification of harmful protists, outperforming traditional methods like quantitative PCR. Notably, ddPCR provides enhanced consistency and reproducibility at it resists PCR inhibitors commonly found in environmental DNA samples. This study aimed to determine the most effective filter material for ddPCR protocols by assessing the reproducibility of species-specific gene copy numbers and filtration time across six filter types: cellulose acetate (CA), mixed cellulose ester (MCE), nylon (NY), polycarbonate (PC), polyethersulfone (PES), and polyvinylidene fluoride (PVDF). The study used two species of Chattonella marina complexes as a case study. Filtration rates were slower for NY, PC, and PVDF filters. Moreover, MCE, NY, PES, and PVDF yielded lower DNA amounts than other filters. Importantly, the CA filter exhibited the lowest variance (38–39%) and the highest determination coefficients (R2 = 0.92–0.96), indicating superior performance. These findings suggest that the CA filter is the most suitable for ddPCR quantification of marine protists, offering quick filtration and reliable reproducibility.

Endogenous virophages are active and mitigate giant virus infection in the marine protist Cafeteria burkhardae

Endogenous viral elements (EVEs) are common genetic passengers in various protists. Some EVEs represent viral fossils, whereas others are still active. The marine heterotrophic flagellate Cafeteria burkhardae contains several EVE types related to the virophage mavirus, a small DNA virus that parasitizes the lytic giant virus CroV. We hypothesized that endogenous virophages may act as an antiviral defense system in protists, but no protective effect of virophages in wild host populations has been shown so far. Here, we tested the activity of virophage EVEs and studied their impact on giant virus replication. We found that endogenous mavirus-like elements (EMALEs) from globally distributed Cafeteria populations produced infectious virus particles specifically in response to CroV infection. However, reactivation was stochastic, often inefficient, and poorly reproducible. Interestingly, only one of eight EMALE types responded to CroV infection, implying that other EMALEs may be linked to different giant viruses. We isolated and cloned several reactivated virophages and characterized their particles, genomes, and infection dynamics. All tested virophages inhibited the production of CroV during coinfection, thereby preventing lysis of the host cultures in a dose-dependent manner. Comparative genomics of different C. burkhardae strains revealed that inducible EMALEs are common and are not linked to specific geographic locations. We demonstrate that naturally occurring virophage EVEs reactivate upon giant virus infection, thus providing a striking example that eukaryotic EVEs can become active under specific conditions. Moreover, our results support the hypothesis that virophages can act as an adaptive antiviral defense system in protists.

Chaotic turnover of rare and abundant species in a strongly interacting model community

The composition of ecological communities varies not only between different locations but also in time. Understanding the fundamental processes that drive species toward rarity or abundance is crucial to assessing ecosystem resilience and adaptation to changing environmental conditions. In plankton communities in particular, large temporal fluctuations in species abundances have been associated with chaotic dynamics. On the other hand, microbial diversity is overwhelmingly sustained by a "rare biosphere" of species with very low abundances. We consider here the possibility that interactions within a species-rich community can relate both phenomena. We use a Lotka-Volterra model with weak immigration and strong, disordered, and mostly competitive interactions between hundreds of species to bridge single-species temporal fluctuations and abundance distribution patterns. We highlight a generic chaotic regime where a few species at a time achieve dominance but are continuously overturned by the invasion of formerly rare species. We derive a focal-species model that captures the intermittent boom-and-bust dynamics that every species undergoes. Although species cannot be treated as effectively uncorrelated in their abundances, the community's effect on a focal species can nonetheless be described by a time-correlated noise characterized by a few effective parameters that can be estimated from time series. The model predicts a nonunitary exponent of the power-law abundance decay, which varies weakly with ecological parameters, consistent with observation in marine protist communities. The chaotic turnover regime is thus poised to capture relevant ecological features of species-rich microbial communities.

Squalene production under oxygen limitation by Schizochytrium sp. S31 in different cultivation systems

The triterpene squalene is widely used in the food, cosmetics and pharmaceutical industries due to its antioxidant, antistatic and anti-carcinogenic properties. It is usually obtained from the liver of deep sea sharks, which are facing extinction. Alternative production organisms are marine protists from the family Thraustochytriaceae, which produce and store large quantities of various lipids. Squalene accumulation in thraustochytrids is complex, as it is an intermediate in sterol biosynthesis. Its conversion to squalene 2,3-epoxide is the first step in sterol synthesis and is heavily oxygen dependent. Hence, the oxygen supply during cultivation was investigated in our study. In shake flask cultivations, a reduced oxygen supply led to increased squalene and decreased sterol contents and yields. Oxygen-limited conditions were applied to bioreactor scale, where squalene accumulation and growth of Schizochytrium sp. S31 was determined in batch, fed-batch and continuous cultivation. The highest dry matter (32.03 g/L) was obtained during fed-batch cultivation, whereas batch cultivation yielded the highest biomass productivity (0.2 g/L*h−1). Squalene accumulation benefited from keeping the microorganisms in the growth phase. Therefore, the highest squalene content of 39.67 ± 1.34 mg/g was achieved by continuous cultivation (D = 0.025 h−1) and the highest squalene yield of 1131 mg/L during fed-batch cultivation. Volumetric and specific squalene productivity both reached maxima in the continuous cultivation at D = 0.025 h−1 (6.94 ± 0.27 mg/L*h−1 and 1.00 ± 0.03 mg/g*h−1, respectively). Thus, the choice of a suitable cultivation method under oxygen-limiting conditions depends heavily on the process requirements.Key points• Measurements of respiratory activity and backscatter light of thraustochytrids• Oxygen limitation increased squalene accumulation in Schizochytrium sp. S31• Comparison of different cultivation methods under oxygen-limiting conditions

Large-scale culturing of the subpolar foraminifera Globigerina bulloides reveals tolerance to a large range of environmental parameters associated to different life-strategies and an extended lifespan.

The subtropical to subpolar planktic foraminifera Globigerina bulloides is a calcifying marine protist, and one of the dominant foraminiferal species of the Nordic Seas. Previously, the relative abundance and shell geochemistry of fossil G. bulloides have been studied for palaeoceanographic reconstructions. There is however a lack of biological observations on the species and a poor understanding of its ecological tolerances, especially for high latitude genotypes. Here, we present observations from the first extensive culturing of G. bulloides under subpolar conditions, including the first low temperature (6-13°C) and variable salinity (30-38) experiments. Carbonate chemistry (pH and [CO3 2-]) was also manipulated. Experimental conditions were chosen to reflect a range of plausible past and future scenarios for the Nordic Seas. We found G. bulloides to be tolerant of environmental conditions well outside their optimal range (<10°C, salinity <33, pH <8). Observed life span was up to three months, which was attributed to a microalgal diet. Two alternative life strategies were employed, whereby individuals either experienced rapid growth and death, or a prolonged lifespan with minimal growth and death via slow decay. We posit this could help explain differences in geochemical signals recorded from different size fractions of fossil specimens used for palaeoceanographic reconstructions.

Distinctive chemotactic responses of three marine herbivore protists to DMSP and related compounds.

Marine planktonic predator-prey interactions occur in microscale seascapes, where diffusing chemicals may act either as chemotactic cues that enhance or arrest predation, or as elemental resources that are complementary to prey ingestion. The phytoplankton osmolyte dimethylsulfoniopropionate (DMSP) and its degradation products dimethylsulfide (DMS) and acrylate are pervasive compounds with high chemotactic potential, but there is a longstanding controversy over whether they act as grazing enhancers or deterrents. Here, we investigated the chemotactic responses of three herbivorous dinoflagellates to point-sourced, microscale gradients of dissolved DMSP, DMS, and acrylate. We found no evidence for acrylate being a chemotactic repellent and observed a weak attractor role of DMS. DMSP behaved as a strong chemoattractor whose potential for grazing facilitation through effects on swimming patterns and aggregation depends on the grazer's feeding mode and ability to incorporate DMSP. Our study reveals that predation models will fail to predict grazing impacts unless they incorporate chemotaxis-driven searching and finding of prey.

Global freshwater distribution of Telonemia protists.

Telonemia are one of the oldest identified marine protists that for most part of their history have been recognized as a distinct incertae sedis lineage. Today, their evolutionary proximity to the SAR supergroup (Stramenopiles, Alveolates, and Rhizaria) is firmly established. However, their ecological distribution and importance as a natural predatory flagellate, especially in freshwater food webs, still remain unclear. To unravel the distribution and diversity of the phylum Telonemia in freshwater habitats, we examined over a thousand freshwater metagenomes from all over the world. In addition, to directly quantify absolute abundances, we analyzed 407 samples from 97 lakes and reservoirs using Catalyzed Reporter Deposition-Fluorescence in situ Hybridization (CARD-FISH). We recovered Telonemia 18S rRNA gene sequences from hundreds of metagenomic samples from a wide variety of habitats, indicating a global distribution of this phylum. However, even after this extensive sampling, our phylogenetic analysis did not reveal any new major clades, suggesting current molecular surveys are near to capturing the full diversity within this group. We observed excellent concordance between CARD-FISH analyses and estimates of abundances from metagenomes. Both approaches suggest that Telonemia are largely absent from shallow lakes and prefer to inhabit the colder hypolimnion of lakes and reservoirs in the Northern Hemisphere, where they frequently bloom, reaching 10%-20% of the total heterotrophic flagellate population, making them important predatory flagellates in the freshwater food web.

Growth Performance of a Newly Isolated and Culturable Thraustochytrid Strain from Sea Squirt Colonies

The world’s oceans and seas host &gt;100 known strains of thraustochytrids, a common group of marine eukaryotic unicellular protists, residing in diverse marine habitats, with many others to be isolated and cultivated. The thraustochytrids have become of considerable industrial interest due to health benefits gained from their high percentages of valuable bioactive compounds, revealing the needs for the isolation of new potential strains. Employing a recently developed isolation methodology (use of cell culture medium), we assess initial culture conditions and growth paces of newly isolated thraustochytrid cells (thraustochytrid sp. BSH), originated from the colonial tunicate Botryllus schlosseri, residing on Helgoland Island, Germany. Cells were cultivated under static versus agitated conditions, along with two inoculation sizes (0.5 × 106 and 1 × 106 cells/dish) and in three vessel types (35 mm Petri dishes and T25 and T75 flasks; containing 3, 5 and 15 mL medium, respectively). Cultures were observed under regular microscopy, confocal microscopy and H&amp;E staining. While cells in all conditions grew fast, results revealed the superiority of agitated cultivation in T75 flasks inoculated with 0.5 × 106 cells/dish (6.41 ± 1.91-fold increase/week). Further, 18S rDNA revealed high similarities (99.5–99.8) of strain BSH to two thraustochytrid strains from Monterey, California (USA), B. schlosseri colonies, elucidating a new understanding of these animals-protists associations.

Kinetic modeling of cell growth and value-added products accumulation of Thraustochytrium striatum

A kinetic mathematical model was developed, calibrated, and validated to describe the fermentation performance of a marine protist, Thraustochytrium striatum. This model consists of equations describing substrate consumption, cell growth, and the accumulation of two important intracellular products (lipids and astaxanthin). Moreover, the substrate inhibition and the consumption of product lipids were also considered in the model. After calibration, this model can describe the fermentation performance of T. striatum with high fidelity (R2 values greater than 0.92) over a broad concentration range of individual (glucose 2.5–50 g/L, cellobiose 2.5–10 g/L, and xylose 2.5–10 g/L) and mixed sugar substrates (glucose/cellobiose/xylose, 10.0/2.5/2.5 g/L and 18.0/4.0/4.0 g/L, respectively). Besides synthetic substrates, this model was also proven to be able to simulate the utilization of natural sugar substrates (i.e., glucose, xylose, and cellobiose) generated from the enzymatic hydrolysate of corn stover. However, the presence of toxic compounds in corn stover hydrolysate might have slightly deviated the model prediction, indicating further improvement of this model is needed when it comes to the simulation of natural sugar substrates containing toxic compounds. This model is anticipated to serve as a fundamental framework to simulate and predict the fermentation performance of marine protist for upscaled applications.

Foraminifera: A Proxy for Ecology, Climate Change and Its Associated Hydro−Environment

Paleocanography, the study of ancient oceans, relies heavily on a wide range of proxies and methodologies to reconstruct past oceanic conditions. Among these tools, foraminifera, microscopic marine organisms with calcium carbonate shells, have played a pivotal role. Over the years, researchers have harnessed the insights provided by foraminifera to unlock the secrets of Earth's oceans throughout geological history. This comprehensive review explores the various ways in which foraminifera have been employed in paleocanography, shedding light on past climate, ocean circulation, ecology, and more. The application of foraminifera represents a sustainable approach with far-reaching implications for environmental, ecological, and geological research. Foraminifera, microscopic marine protists characterized by calcium carbonate shells, serve as valuable indicators of past and present environmental conditions. They are instrumental in the fields of paleoecology, stratigraphy, environmental impact studies, and ocean acidification research. This abstract highlights the sustainable aspect of using foraminifera as bioindicators to assess the health of marine ecosystems, monitor climate change, and investigate the impact of human activities. By providing insights into the past and present, foraminifera contribute to informed decision-making in conservation, resource management, and addressing critical environmental challenges. Their application stands as a testament to the enduring value of these microorganisms in understanding and safeguarding our planet's natural resources and ecosystems, exemplifying the sustainability of scientific research driven by a profound respect for the Earth's history and future well-being.