The green seaweed Ulva: tomorrow's "wheat of the sea" in foods, feeds, nutrition, and biomaterials.

Ulva, a genus of green macroalgae commonly known as sea lettuce, has long been recognized for its nutritional benefits for food and feed. As the demand for sustainable food and feed sources continues to grow, so does the interest in alternative, plant-based protein sources. With its abundance along coastal waters and high protein content, Ulva spp. have emerged as promising candidates. While the use of Ulva in food and feed has its challenges, the utilization of Ulva in other industries, including in biomaterials, biostimulants, and biorefineries, has been growing. This review aims to provide a comprehensive overview of the current status, challenges and opportunities associated with using Ulva in food, feed, and beyond. Drawing on the expertise of leading researchers and industry professionals, it explores the latest knowledge on Ulva's nutritional value, processing methods, and potential benefits for human nutrition, aquaculture feeds, terrestrial feeds, biomaterials, biostimulants and biorefineries. In addition, it examines the economic feasibility of incorporating Ulva into aquafeed. Through its comprehensive and insightful analysis, including a critical review of the challenges and future research needs, this review will be a valuable resource for anyone interested in sustainable aquaculture and Ulva's role in food, feed, biomaterials, biostimulants and beyond.

Temporal force governs the microbial assembly associated with Ulva fasciata (Chlorophyta) from an integrated multi-trophic aquaculture system.

Ulva spp., one of the most important providers of marine ecosystem services, has gained substantial attention lately in both ecological and applicational aspects. It is known that macroalgae and their associated microbial community form an inseparable unit whose intimate relationship can affect the wellbeing of both. Different cultivation systems, such as integrated multi-trophic aquaculture (IMTA), are assumed to impact Ulva bacterial community significantly in terms of compositional guilds. However, in such a highly dynamic environment, it is crucial to determine how the community dynamics change over time. In the current study, we characterized the microbiota associated with Ulva fasciata grown as a biofilter in an IMTA system in the Gulf of Aqaba (Eilat, Israel) over a developmental period of 5 weeks. The Ulva-associated microbial community was identified using the 16S rRNA gene amplicon sequencing technique, and ecological indices were further analyzed. The Ulva-associated microbiome revealed a swift change in composition along the temporal succession, with clusters of distinct communities for each timepoint. Proteobacteria, Bacteroidetes, Planctomycetes, and Deinococcus-Thermus, the most abundant phyla that accounted for up to 95% of all the amplicon sequence variants (ASVs) found, appeared in all weeks. Further analyses highlighted microbial biomarkers representing each timepoint and their characteristics. Finally, the presence of highly abundant species in Ulva microbiota yet underestimated in previous research (such as phyla Deinococcus-Thermus, families Saprospiraceae, Thiohalorhabdaceae, and Pirellulaceae) suggests that more attention should be paid to the temporal succession of the assembly of microbes inhabiting macroalgae in aquaculture, in general, and IMTA, in particular. Characterizing bacterial communities associated with Ulva fasciata from an IMTA system provided a better understanding of their associated microbial dynamics and revealed this macroalgae's adaptation to such a habitat.

Ecological insights into the resilience of marine plastisphere throughout a storm disturbance.

Among numerous research about marine plastisphere, the community living on the surface of plastic debris, little attention was given to the ecological mechanisms governing prokaryotes compared to eukaryotes, and even less focused on their resilience in a changing climate with more storm prevalence. Our current research recruited an integrated approach involving community succession across temporal dimension, ecological mechanisms that govern the assembly, and resilience to environmental perturbations to highlight the ecology of different kingdoms in the plastisphere. Towards this goal, we examined the succession of the prokaryotic and eukaryotic communities on artificial plastic nets in a sidestream of seawater from the Gulf of Aqaba over 35 days. A robust local storm enabled investigation of the alterations before, during, and after this disturbance, aiming at the community's potential to recover. Data from 16S and 18S rRNA sequencing and microscopic analyses decrypted the plastisphere diversity, community assembly, and stochasticity, followed by further analyses of functional and co-occurrence networks for the prokaryotic group. Prokaryotic and eukaryotic communities underwent exact opposite ecological mechanisms. While determinism driven by a robust environmental selection dictated the prokaryotic community assembly, stochasticity prevailed when this condition was relaxed. Interestingly, resilience against disturbance was observed in prokaryotes but not in eukaryotes. The decrease in compositional, functional diversity and network complexity in the prokaryotic community was reversed, presumably due to the niche specification process and high dispersal. Niche specification following perturbation was evident in some bacteria by selected functions associated with plastic degradation, stress response, and antibiotic resistance. On the contrary, eukaryotes decreased in diversity and were dominated by the commonly found Chlorophyta towards the later successional period. Novel findings on the ecology of marine plastisphere during perturbation encourage the integration of this aspect into prediction research.

Spatial Succession Underlies Microbial Contribution to Food Digestion in the Gut of an Algivorous Sea Urchin.

Dietary influence on the microbiome in algivorous sea urchins such as Tripneustes gratilla elatensis suggests a bacterial contribution to the digestion of fiber-rich seaweed. An ecological insight into the spatial arrangement in the gut bacterial community will improve our knowledge of host-microbe relations concerning the involved taxa, their metabolic repertoire, and the niches of activity. Toward this goal, we investigated the bacterial communities in the esophagus, stomach, and intestine of Ulva-fed sea urchins through 16S rRNA amplicon sequencing, followed by the prediction of their functional genes. We revealed communities with distinct features, especially those in the esophagus and intestine. The esophageal community was less diverse and was poor in food digestive or fermentation genes. In contrast, bacteria that can contribute to the digestion of the dietary Ulva were common in the stomach and intestine and consisted of genes for carbohydrate decomposition, fermentation, synthesis of short-chain fatty acids, and various ways of N and S metabolism. Bacteroidetes and Firmicutes were found as the main phyla in the gut and are presumably also necessary in food digestion. The abundant sulfate-reducing bacteria in the stomach and intestine from the genera Desulfotalea, Desulfitispora, and Defluviitalea may aid in removing the excess sulfate from the decomposition of the algal polysaccharides. Although these sea urchins were fed with Ulva, genes for the degradation of polysaccharides of other algae and plants were present in this sea urchin gut microbiome. We conclude that the succession of microbial communities along the gut obtained supports the hypothesis on bacterial contribution to food digestion. IMPORTANCE Alga grazing by the sea urchin Tripneustes gratilla elatensis is vital for nutrient recycling and constructing new reefs. This research was driven by the need to expand the knowledge of bacteria that may aid this host in alga digestion and their phylogeny, roles, and activity niches. We hypothesized alterations in the bacterial compositional structure along the gut and their association with the potential contribution to food digestion. The current spatial insight into the sea urchin's gut microbiome ecology is novel and reveals how distinct bacterial communities are when distant from each other in this organ. It points to keynote bacteria with genes that may aid the host in the digestion of the complex sulfated polysaccharides in dietary Ulva by removing the released sulfates and fermentation to provide energy. The gut bacteria's genomic arsenal may also help to gain energy from diets of other algae and plants.

Isolation of bacteria capable of growth with 2-methylisoborneol and geosmin as the sole carbon and energy sources.

Using a relatively simple enrichment technique, geosmin and 2-methylisoborneol (MIB)-biodegrading bacteria were isolated from a digestion basin in an aquaculture unit. Comparison of 16S rRNA gene sequences affiliated one of the three isolates with the Gram-positive genus Rhodococcus, while the other two isolates were found to be closely related to the Gram-negative family Comamonadaceae (Variovorax and Comamonas). Growth rates and geosmin and MIB removal rates by the isolates were determined under aerated and nonaerated conditions in mineral medium containing either of the two compounds as the sole carbon and energy source. All isolates exhibited their fastest growth under aerobic conditions, with generation times ranging from 3.1 to 5.7 h, compared to generation times of up to 19.1 h in the nonaerated flasks. Incubation of the isolates with additional carbon sources caused a significant increase in their growth rates, while removal rates of geosmin and MIB were significantly lower than those for incubation with only geosmin or MIB. By fluorescence in situ hybridization, members of the genera Rhodococcus and Comamonas were detected in geosmin- and MIB-enriched sludge from the digestion basin.

Carbohydrate-Active Enzymes of a Novel Halotolerant Alkalihalobacillus Species for Hydrolysis of Starch and Other Algal Polysaccharides.

Halotolerant bacteria capable of starch hydrolysis by their amylases will benefit various industries, specifically since the hydrolytic activity of current industrial amylases is inhibited or even absent in salt-rich or alkaline environments. Seeking novel enzymes, we analyzed the entire genome content of a marine bacterium isolated from the gut of sea urchins to compare it against other bacterial genomes. Conditions underlying α-amylase activity were examined in vitro at various salinities (0 to 4%) and temperatures (25°C to 37°C). Genomic analyses revealed the isolated bacterium as a new species of Alkalihalobacillus. Comparative analysis of the contents of carbohydrate-active enzymes revealed various α-amylases, each with its respective carbohydrate-binding module for starch hydrolysis. Functional analysis identified the hydrolysis of starch and the maltooligosaccharides maltose and dextrin into d- and UDP-glucose. The fastest growth and α-amylase production occurred at 3% salinity at a temperature of 30°C. The Alkalihalobacillus sp. consists of exclusive contents of α-amylases and other enzymes that may be valuable in the hydrolysis of the algal polysaccharides cellulose and laminarin. IMPORTANCE Toward the discovery of novel carbohydrate-active enzymes that may be useful in the hydrolysis of starch, we examined a halotolerant bacterial isolate of Alkalihalobacillus sp. regarding its genomic content and conditions underlying the production of active α-amylases. The production of α-amylases was measured in bacterial cultures at relatively high temperature (37°C) and salinity (4%). The Alkalihalobacillus sp. revealed an exclusive content of amylases and other carbohydrate-active enzymes compared to other relevant bacteria. These enzymes may be valuable for the hydrolysis of algal polysaccharides. The enzymatic cascade of the Alkalihalobacillus sp. for starch metabolism allows polysaccharide degradation into monosugars while preventing the accumulation of intermediate inhibitors of maltose or dextrin.

Fishmeal replacement by periphyton reduces the fish in fish out ratio and alimentation cost in gilthead sea bream Sparus aurata.

Aquaculture threatens natural resources by fishing down the sea to supply fishmeal. Alternative protein sources in aquafeeds can provide a solution, particularly those that are waste from other operations and thereby reduce feed production costs. Toward this goal, we examined the waste biomass of marine periphyton from biofilters of an integrated multi-trophic aquaculture (IMTA) system as a replacement for fishmeal in diets of gilthead seabream (Sparus aurata). Four isoproteic (41%) and isolipidic (16.7%) aquafeeds were formulated with increased content of periphyton and a corresponding decrease in fishmeal from 20 to 15, 10, or 0%. The growth and biochemical content of seabream fingerlings (initial body weight 10 g) were examined over 132 days. Replacing 50% of fishmeal by waste periphyton improved feed conversion ratio (1.2 vs. 1.35 in the control diet) without harming fish growth. The complete replacement of fishmeal with periphyton resulted in 15% slower growth but significantly higher protein content in the fish flesh (59 vs. 52% in the control diet). Halving fishmeal content reduced feed cost by US$ 0.13 kg-1 feed and saved 30% in the cost of conversion of feed to fish biomass (US$ 0.58 kg-1 produced fish vs. $0.83 in the control diet). Finally, the total replacement of fishmeal by waste periphyton in the diet reduced the fish in-fish out ratio to below 1 (0.5-0.9) as compared to 1.36 in the control diet. Replacing fishmeal with on-farm produced periphyton minimizes aquaculture footprint through the removal of excess nutrients in effluents and the use of waste biomass to reduce the 'fish in' content in aquafeeds and fish production costs. The present study demonstrates the great practical potential of this dual use of marine periphyton in enhancing the circular economy concept in sustainable fish production.

Mono-specific algal diets shape microbial networking in the gut of the sea urchin Tripneustes gratilla elatensis.

BACKGROUND: Algivorous sea urchins can obtain energy from a diet of a single algal species, which may result in consequent changes in their gut microbe assemblies and association networks. METHODS: To ascertain whether such changes are led by specific microbes or limited to a specific region in the gut, we compared the microbial assembly in the three major gut regions of the sea urchin Tripneustes gratilla elatensis when fed a mono-specific algal diet of either Ulva fasciata or Gracilaria conferta, or an algal-free diet. DNA extracts from 5 to 7 individuals from each diet treatment were used for Illumina MiSeq based 16S rRNA gene sequencing (V3-V4 region). Niche breadth of each microbe in the assembly was calculated for identification of core, generalist, specialist, or unique microbes. Network analyzers were used to measure the connectivity of the entire assembly and of each of the microbes within it and whether it altered with a given diet or gut region. Lastly, the predicted metabolic functions of key microbes in the gut were analyzed to evaluate their potential contribution to decomposition of dietary algal polysaccharides. RESULTS: Sea urchins fed with U. fasciata grew faster and their gut microbiome network was rich in bacterial associations (edges) and networking clusters. Bacteroidetes was the keystone microbe phylum in the gut, with core, generalist, and specialist representatives. A few microbes of this phylum were central hub nodes that maintained community connectivity, while others were driver microbes that led the rewiring of the assembly network based on diet type through changes in their associations and centrality. Niche breadth agreed with microbes' richness in genes for carbohydrate active enzymes and correlated Bacteroidetes specialists to decomposition of specific polysaccharides in the algal diets. CONCLUSIONS: The dense and well-connected microbial network in the gut of Ulva-fed sea urchins, together with animal's rapid growth, may suggest that this alga was most nutritious among the experimental diets. Our findings expand the knowledge on the gut microbial assembly in T. gratilla elatensis and strengthen the correlation between microbes' generalism or specialism in terms of occurrence in different niches and their metabolic arsenal which may aid host nutrition.

Light and Primary Production Shape Bacterial Activity and Community Composition of Aerobic Anoxygenic Phototrophic Bacteria in a Microcosm Experiment.

Phytoplankton is a key component of aquatic microbial communities, and metabolic coupling between phytoplankton and bacteria determines the fate of dissolved organic carbon (DOC). Yet, the impact of primary production on bacterial activity and community composition remains largely unknown, as, for example, in the case of aerobic anoxygenic phototrophic (AAP) bacteria that utilize both phytoplankton-derived DOC and light as energy sources. Here, we studied how reduction of primary production in a natural freshwater community affects the bacterial community composition and its activity, focusing primarily on AAP bacteria. The bacterial respiration rate was the lowest when photosynthesis was reduced by direct inhibition of photosystem II and the highest in ambient light condition with no photosynthesis inhibition, suggesting that it was limited by carbon availability. However, bacterial assimilation rates of leucine and glucose were unaffected, indicating that increased bacterial growth efficiency (e.g., due to photoheterotrophy) can help to maintain overall bacterial production when low primary production limits DOC availability. Bacterial community composition was tightly linked to light intensity, mainly due to the increased relative abundance of light-dependent AAP bacteria. This notion shows that changes in bacterial community composition are not necessarily reflected by changes in bacterial production or growth and vice versa. Moreover, we demonstrated for the first time that light can directly affect bacterial community composition, a topic which has been neglected in studies of phytoplankton-bacteria interactions.IMPORTANCE Metabolic coupling between phytoplankton and bacteria determines the fate of dissolved organic carbon in aquatic environments, and yet how changes in the rate of primary production affect the bacterial activity and community composition remains understudied. Here, we experimentally limited the rate of primary production either by lowering light intensity or by adding a photosynthesis inhibitor. The induced decrease had a greater influence on bacterial respiration than on bacterial production and growth rate, especially at an optimal light intensity. This suggests that changes in primary production drive bacterial activity, but the effect on carbon flow may be mitigated by increased bacterial growth efficiencies, especially of light-dependent AAP bacteria. Bacterial activities were independent of changes in bacterial community composition, which were driven by light availability and AAP bacteria. This direct effect of light on composition of bacterial communities has not been documented previously.

2-Methylisoborneol and geosmin uptake by organic sludge derived from a recirculating aquaculture system.

In a previous study on a recirculating fish culture system, levels of geosmin and 2-methylisoborneol were found to decrease when culture water was recirculated through the anaerobic sludge digestion treatment stage of the system. This finding led us to the present study in which the geosmin and 2-methylisoborneol removal capacity of the sludge derived from this treatment stage was examined in vitro. It was found that reduction of off-flavor compounds by the sludge was mediated by both chemical/physical sorption and biological degradation. At geosmin and 2-methylisoborneol concentrations within the range of those experienced in fish culture systems, chemical/physical sorption by the sludge was found to account for a 93% reduction in geosmin and a 79% reduction in 2-methylisoborneol from the overlying water within 48h of incubation. Combined with the biological degradation taking place in the sludge, a complete removal of these compounds from the water phase occurred within 9 days of incubation. By means of repeated washing of the geosmin and 2-methylisoborneol contaminate sludge with clean water, relatively small amounts of these compounds were released from the sludge, a possible indication for the fact that absorption, rather than adsorption, underlies the chemical/physical removal process.