Inclusion of dietary Ulva ohnoi 5% modulates Solea senegalensis immune response during Photobacterium damselae subsp. piscicida infection.

Macroalgae represent valuable sources of functional ingredients for fish diets, and the influence of supplemented aquafeeds on growth performance has been studied for some fish and seaweed species. In the present work, the potential immunomodulation exerted by U. ohnoi (5%) as dietary ingredient was investigated in Senegalese sole. After feeding with the experimental diets for 90 d, fish immune response before and after challenge with Photobacterium damselae subsp. piscicida (Phdp) was assessed. In absence of infection, systemic immune response was not modified by 5% U. ohnoi dietary inclusion for 90 d. Thus, no differences in liver and head kidney immune gene transcription or serum lysozyme, peroxidase, antiprotease and complement activities were observed based on the diet received by Senegalese sole specimens. Regarding mucosal immune parameters, no changes in gene transcription were detected in the skin and gills, whilst only tnf, cd4 and cd8 were significantly up-regulated in the intestine of fish fed with U. ohnoi, compared to the values obtained with control diet. On the contrary, when S. senegalensis specimens were challenged with Phdp, modulation of the immune response consisting in increased transcription of genes encoding complement (c1q4, c3, c9), lysozyme g (lysg), tumor necrosis factor alpha (tnfα) as well as those involved in the antioxidant response (gpx, sodmn) and iron metabolism (ferrm, hamp-1) was observed in the liver of fish fed with U. ohnoi. In parallel, decreased inflammatory cytokine and complement encoding gene transcription was displayed by the spleen of fish receiving the algal diet. Though mortality rates due to Phdp challenge were not affected by the diet received, lower pathogen loads were detected in the liver of soles receiving U. ohnoi diet. Further research to investigate the effects of higher inclusion levels of this seaweed in fish diets, feeding during short periods as wells as to assess the response against other pathogens needs to be carried out.

Partial Characterization of Protease Inhibitors of Ulva ohnoi and Their Effect on Digestive Proteases of Marine Fish.

This piece of research evaluates the presence of protease inhibitors in the macroalga Ulva ohnoi and provides an initial overview of their mode of action. The ability of Ulva protease inhibitors to inhibit digestive proteases of three marine fish species, as well as their capacity to hamper the hydrolysis of a reference protein by those fish proteases, were assessed. In addition, thermal stability and the mode of inhibition on trypsin and chymotrypsin were also studied. Dose-response inhibition curves and in vitro protein hydrolysis assays revealed a noticeable inhibition of fish enzymes when Ulva concentration increased in the assay. The thermal treatment of Ulva reduced markedly the inhibitory effect on fish digestive protease. Finally, Lineweaver-Burk plots indicated that trypsin and chymotrypsin inhibition consisted of a mixed-type inhibition mechanism in which the inhibitory effect depends on Ulva concentration. Overall, the results confirmed the presence of protease inhibitors in Ulva, though heat treatment was enough for inactivating these compounds.

Phytoremediation potential of Ulva ohnoi (Chlorophyta): Influence of temperature and salinity on the uptake efficiency and toxicity of cadmium.

Ulva ohnoi is a green macroalga with fast growth and high rates of nitrogen and phosphorus absorption. Recently, this species has been recorded in several places with record green tide formation in some of them. Using molecular tools, we herein report the first occurrence of this species in Brazil and demonstrate its potential for phytoremediation in typical environmental concentrations of Cd (0.625-15 µg L-1). Similarly, the effects of physicochemical parameters (salinity and temperature) on the toxicity and uptake efficiency of this species were evaluated. Molecular analysis of two sequences (1141 bp) obtained corroborates another 34 sequences for U. ohnoi obtained from GenBank. The addition of Cd in the medium affected photosynthetic parameters and reduced growth rate. U. ohnoi showed resistance to Cd when cultivated at 18 °C, S15 and 18-25 °C, S35, at concentrations between 0.625 and 2.5 µg. L-1 of Cd; yet, positive growth rate was maintained. Dose-dependent accumulation was observed in all combinations of factors used with a maximum value of 4.20 µg Cd per gram of dry seaweed at 15 µg. L-1 of Cd at 18 °C and S35. Maximum value of the concentration factor was 81.3 ±â€¯1.1% of Cd added at the concentration of 0.625 µg. L-1 to S15 and 18 °C. Our results demonstrate the potential of using U. ohnoi in the phytoremediation of Cd in saltwater or brackish water.

Examination of prezygotic and postzygotic isolating barriers in tropical Ulva (Ulvophyceae, Chlorophyta): evidence for ongoing speciation.

Phylogenetic clades based on DNA sequences such as the chloroplast rbcL gene and the nuclear ITS region are frequently used to delimit algal species. However, these molecular markers cannot accurately delimit boundaries among some Ulva species. Although Ulva reticulata and Ulva ohnoi occasionally bloom in tropical to warm-temperate regions and are clearly distinguishable by their reticulate or plain blade morphology, they have few or no sequence divergences in these molecular markers and form a monophyletic clade. In this study, to clarify the speciation and species delimitation in the U. reticulata-ohnoi complex clade, reproductive relationships among several sexual strains from the Philippines and Japan including offspring that originated from the type specimen of U. ohnoi were examined by culturing and hybridization in addition to the ITS-based analysis. As a result, both prezygotic and postzygotic reproductive isolation were revealed to occur between genetically perforated U. reticulata and imperforate U. ohnoi. They were also separated on the basis of sequence analysis of the ITS region. That strongly supports that the two taxa are independent biological species. Although no prezygotic barrier among the Philippine and Japanese strains of U. reticulata was observed, unexpectedly zoospores produced by hybrid sporophytes in some of their combinations mostly failed to develop, indicating partial formation of a postzygotic barrier despite a 0.2% divergence in the ITS sequence. These findings suggest speciation is still ongoing in U. reticulata.

Modelling bioremediation of contaminated effluents by Ulva ohnoi. - A predictive perspective.

Ulva spp. are tolerant to salinity variations and exhibit easy acclimation, playing an essential role in the depollution of aquatic ecosystems precisely due to their high efficiency in absorbing and accumulating nutrients. For this reason, Ulva spp. becomes an attractive solution for recovering areas that suffer the impacts of problems such as the eutrophication of anthropogenic origin. In addition to being a promising alternative for the blue bioeconomy, it can contribute to the sustainability of economic activities in coastal areas. Therefore, the present study aimed to develop and elucidate the behavior of Ulva ohnoi using predictive surface response models. The algae were grown under different concentrations of nutrient and salinity levels, as predicted by the experimental design, and it was evaluated according to the potential of the biomass to absorb the nutrients, as well as its photosynthetic performance and biochemical parameters. Our study confirmed the high efficiency and preference of Ulva ohnoi in the absorption of nitrogen dissolved in the medium in the form of NH4+ and that salinity is an essential factor in the dynamics and speed of ammonium absorption. The absorption of orthophosphate by U. ohnoi is reverted to the culture medium when subjected to long-term cultivation. This process was more intense because of low salinity, even at conditions of availability of the compound. The 3D-models of response surfaces elucidate the behavior of Ulva ohnoi, attributing a correlation between nutrient availability and salinity and the biological behavior of the species. In view of what is exposed by these models, as well as the effects of saline distribution along the Lagoon, the following regions of the lagoon are suggested: Center-North, Center and South - as potential areas for the implementation of bioremediation projects with Ulva ohnoi.

Holistic understanding of the response of grapevines to foliar application of seaweed extracts.

Viticulture is highly dependent on phytochemicals to maintain good vineyard health. However, to reduce their accumulation in the environment, green regulations are driving the development of eco-friendly strategies. In this respect, seaweeds have proven to be one of the marine resources with the highest potential as plant protective agents, representing an environmentally-friendly alternative approach for sustainable wine production. The current work follows an interdisciplinary framework to evaluate the capacity of Ulva ohnoi and Rugulopteryx okamurae seaweeds to induce defense mechanisms in grapevine plants. To our knowledge, this is the first study to evaluate Rugulopteryx okamurae as a biostimulator . This macroalgae is relevant since it is an invasive species on the Atlantic and Mediterranean coast causing incalculable economic and environmental burdens. Four extracts (UL1, UL2, RU1 and RU2 developed from Ulva and Rugulopteryx, respectively) were foliar applied to Tempranillo plants cultivated under greenhouse conditions. UL1 and RU2 stood out for their capacity to induce defense genes, such as a PR10, PAL, STS48 and GST1, mainly 24 hours after the first application. The increased expression level of these genes agreed with i) an increase in trans-piceid and trans-resveratrol content, mainly in the RU2 treated leaves, and, ii) an increase in jasmonic acid and decrease in salicylic acid. Moreover, an induction of the activity of the antioxidant enzymes was observed at the end of the experiment, with an increase in superoxide dismutase and catalase in the RU2-treated leaves in particular. Interestingly, while foliar fungal diversity was not influenced by the treatments, alga extract amendment modified fungal composition, RU2 application enriching the content of various groups known for their biocontrol activity. Overall, the results evidenced the capacity of Rugulopteryx okamurae for grapevine biostimulation, inducing the activation of several secondary metabolite pathways and promoting the abundance of beneficial microbiota involved in grapevine protection. While further studies are needed to unravel the bioactive compound(s) involved, including conducting field experiments etc., the current findings are the first steps towards the inclusion of Rugulopteryx okamurae in a circular scheme that would reduce its accumulation on the coast and benefit the viticulture sector at the same time.

Evaluation of the Inclusion of the Green Seaweed Ulva ohnoi as an Ingredient in Feeds for Gilthead Sea Bream (Sparus aurata) and European Sea Bass (Dicentrarchus labrax).

This study evaluated the use of Ulva ohnoi as an ingredient in feeds for aquaculture in three different experiments. Experiment 1 was oriented to confirm the negative effect of U. ohnoi on fish digestion. Experiment 2 assessed the effect on growth, feed efficiency, and immune status of juvenile sea bass (Dicentrarchus labrax) fed on diets including U. ohnoi, previously treated or not with carbohydrases used to partially hydrolyze indigestible polysaccharides. Experiment 3 was aimed to evaluate the potential protective effect of U. ohnoi on the oxidative status of sea bream (Sparus aurata) challenged by the consumption of a feed formulated with the oil fraction completely oxidized. Results show a negligible effect of U. ohnoi meal on protein digestion when included in feeds at levels of 10% or less. Moreover, results of growth and feed use evidenced the possibility of using up to 5% inclusion of algal meal in feeds without adverse effects on the zootechnical parameters, while the enzyme pretreatment was ineffective to improve its nutritional use. Finally, the inclusion of U. onhoi in feeds determined both an immunostimulatory effect, evidenced by an increase in skin mucus lysozyme in the two mentioned fish species, and a positive influence on the oxidative metabolism of seabream when fed on a diet including rancid oil.

Massive Ulva Green Tides Caused by Inhibition of Biomass Allocation to Sporulation.

The green seaweed Ulva spp. constitute major primary producers in marine coastal ecosystems. Some Ulva populations have declined in response to ocean warming, whereas others cause massive blooms as a floating form of large thalli mostly composed of uniform somatic cells even under high temperature conditions-a phenomenon called "green tide". Such differences in population responses can be attributed to the fate of cells between alternative courses, somatic cell division (vegetative growth), and sporic cell division (spore production). In the present review, I attempt to link natural population dynamics to the findings of physiological in vitro research. Consequently, it is elucidated that the inhibition of biomass allocation to sporulation is an important key property for Ulva to cause a huge green tide.

Evaluating bloom potential of the green-tide forming alga Ulva ohnoi under ocean acidification and warming.

The occurrence of green-tides, whose bloom potential may be increased by various human activities and biogeochemical process, results in enormous economic losses and ecosystem collapse. In this study, we investigated the ecophysiology of the subtropical green-tide forming alga, Ulva ohnoi complex (hereafter: U. ohnoi), under simulated future ocean conditions in order to predict its bloom potential using photosynthesis and growth measurements, and stable isotope analyses. Our mesocosm system included four experimental conditions that simulated the individual and combined effects of elevated CO2 and temperature, namely control (450 µatm CO2 & 20 °C), acidification (900 µatm CO2 & 20 °C), warming (450 µatm CO2 & 25 °C), and greenhouse (900 µatm CO2 & 25 °C). Photosynthetic electron transport rates (rETR) increased significantly under acidification conditions, but net photosynthesis and growth were not affected. In contrast, rETR, net photosynthesis, and growth all decreased significantly under elevated temperature conditions (i.e. both warming and greenhouse). These results represent the imbalance of energy metabolism between electron transport and O2 production that may be expected under ocean acidification conditions. This imbalance appears to be related to carbon and nitrogen assimilation by U. ohnoi. In particular, 13C and 15N discrimination data suggest U. ohnoi prefers CO2 and NH4+ over HCO3- and NO3- as sources of carbon and nitrogen, respectively, and this results in increased N content in the thallus under ocean acidification conditions. Together, our results suggest a trade-off in which the bloom potential of U. ohnoi could increase under ocean acidification due to greater N accumulation and through the saving of energy during carbon and nitrogen metabolism, but that elevated temperatures could decrease U. ohnoi's bloom potential through a decrease in photosynthesis and growth.

Mitochondrial and chloroplast genome sequences of Ulva ohnoi, a green-tide-forming macroalga in the Southern coastal regions of Japan.

Ulva is a green macroalga often causing a macroalgal bloom, 'green tide'. Ulva ohnoi is a major species composing the green tide of the southern coastal regions of Japan. Here, we sequenced the complete mitochondrial and chloroplast genomes of the authentic strain of U. ohnoi. The mitochondrial and chloroplast genomes were of 65,326 bp and 103,313 bp, respectively, and the gene content was highly conserved in the Ulva species. The phylogenetic analyses using mitochondrial or chloroplast proteins represented the same topology with high supporting values. These results show that mitochondrial and chloroplast genomes can be used as reliable phylogenetic markers.