Identification of reproductive sex-biased gene expression in Asparagopsis taxiformis (lineage 6) gametophytes.

The sub-tropical red seaweed Asparagopsis taxiformis is of significant interest due to its ability to store halogenated compounds, including bromoform, which can mitigate methane production in ruminants. Significant scale-up of aquaculture production of this seaweed is required; however, relatively little is known about the molecular mechanisms that control fundamental physiological processes, including the regulatory factors that determine sexual dimorphism in gametophytes. In this study, we used comparative RNA-sequencing analysis between different morphological parts of mature male and female A. taxiformis (lineage 6) gametophytes that resulted in greater number of sex-biased gene expression in tips (containing the reproductive structures for both sexes), compared with the somatic main axis and rhizomes. Further comparative RNA-seq against immature tips was used to identify 62 reproductive sex-biased genes (59 male-biased, 3 female-biased). Of the reproductive male-biased genes, 46% had an unknown function, while others were predicted to be regulatory factors and enzymes involved in signaling. We found that bromoform content obtained from female samples (8.5 ± 1.0 mg·g-1 dry weight) was ~10% higher on average than that of male samples (6.5 ± 1.0 mg·g-1 dry weight), although no significant difference was observed (p > 0.05). There was also no significant difference in the marine bromoform biosynthesis locus gene expression. In summary, our comparative RNA-sequencing analysis provides a first insight into the potential molecular factors relevant to gametogenesis and sexual differentiation in A. taxiformis, with potential benefits for identification of sex-specific markers.

Symposium review: Effective nutritional strategies to mitigate enteric methane in dairy cattle.

Intensive research in the past decade has resulted in a better understanding of factors driving enteric methane (CH4) emissions in ruminants. Meta-analyses of large databases, developed through the GLOBAL NETWORK project, have identified successful strategies for mitigation of CH4 emissions. Methane inhibitors, alternative electron sinks, vegetable oils and oilseeds, and tanniferous forages are among the recommended strategies for mitigating CH4 emissions from dairy and beef cattle and small ruminants. These strategies were also effective in decreasing CH4 emissions yield and intensity. However, a higher inclusion rate of oils may negatively affect feed intake, rumen function, and animal performance, specifically milk components in dairy cows. In the case of nitrates (electron sinks), concerns with animal health may be impeding their adoption in practice, and potential emission trade-offs have to be considered. Tannins and tanniferous forages may have a negative effect on nutrient digestibility, and more research is needed to confirm their effects on overall animal performance in long-term experiments with high-producing animals. A meta-analysis of studies with dairy cows fed the CH4 inhibitor 3-nitrooxypropanol (3-NOP) at the Pennsylvania State University showed (1) a consistent 28 to 32% decrease in daily CH4 emissions or emissions yield and intensity; (2) no effect on dry matter intake, milk production, body weight, or body weight change, and a slight increase in milk fat concentration and yield (0.19 percentage units and 90 g/d, respectively); 3-NOP also appears to increase milk urea nitrogen concentration; (3) an exponential decrease in the mitigation effect of the inhibitor with increasing its dose (from 40 to 200 mg/kg of feed dry matter, corresponding to 3-NOP intake of 1 to 4.8 g/cow per day); and (4) a potential decrease in the efficacy of 3-NOP over time, which needs to be further investigated in long-term, full-lactation or multiple-lactation studies. The red macroalga Asparagopsis taxiformis has a strong CH4 mitigation effect, but studies are needed to determine its feasibility, long-term efficacy, and effects on animal production and health. We concluded that widespread adoption of mitigation strategies with proven effectiveness by the livestock industries will depend on cost, government policies and incentives, and willingness of consumers to pay a higher price for animal products with decreased carbon footprint.

Asparagopsis Genus: What We Really Know About Its Biological Activities and Chemical Composition.

Although the genus Asparagopsis includes only two taxonomically accepted species, the published literature is unanimous about the invasive nature of this genus in different regions of the globe, and about the availability of large amounts of biomass for which it is important to find a commercial application. This review shows that extracts from Asparagospsis species have already been evaluated for antioxidant, antibacterial, antifungal, antiviral, antifouling, cytotoxic, antimethanogenic and enzyme-inhibitory activity. However, the tables presented herein show, with few exceptions, that the activity level displayed is generally low when compared with positive controls. Studies involving pure compounds being identified in Asparagopsis species are rare. The chemical compositions of most of the evaluated extracts are unknown. At best, the families of the compounds present are suggested. This review also shows that the volatile halogenated compounds, fatty acids and sterols that are biosynthesized by the Asparagopsis species are relatively well known. Many other non-volatile metabolites (halogen compounds, flavonoids, other phenolic compounds) seem to be produced by these species, but their chemical structures and properties haven'been investigated. This shows how much remains to be investigated regarding the secondary-metabolite composition of these species, suggesting further studies following more targeted methodologies.

Effects of the macroalga Asparagopsis taxiformis and oregano leaves on methane emission, rumen fermentation, and lactational performance of dairy cows.

Asparagopsis taxiformis (AT) is a source of multiple halogenated compounds and, in a limited number of studies, has been shown to decrease enteric CH4 emission in vitro and in vivo. Similarly, oregano has been suggested as a potential CH4 mitigating agent. This study consisted of 2 in vitro and 2 in vivo experiments. Experiment (Exp.) 1 was aimed at establishing the effect of AT on CH4 emission in vitro. Two experiments (Exp. 2 and 3) with lactating dairy cows were conducted to determine the antimethanogenic effect of AT and oregano (Exp. 3) in vivo. Another experiment (Exp. 4) was designed to investigate stability of bromoform (CHBr3) in AT over time. In Exp. 3, 20 Holstein cows were used in a replicated 4 × 4 Latin square design with four 28-d periods. Treatments were basal diet (control) or basal diet supplemented with (dry matter basis) 0.25% AT (LowAT), 0.50% AT (HighAT), or 1.77% oregano (Origanum vulgare L.) leaves. Enteric gas emissions were measured using the GreenFeed system (C-Lock Inc., Rapid City, SD), and rumen samples were collected for fermentation analysis using the ororuminal technique. In Exp.1 (in vitro), relative to the control, AT (at 1% dry matter basis, inclusion rate) decreased CH4 yield by 98%. In Exp. 3, HighAT decreased average daily CH4 emission and CH4 yield by 65% and 55%, respectively, in experimental periods 1 and 2, but had no effect in periods 3 and 4. The differential response to AT among experimental periods was likely a result of a decrease in CHBr3 concentration in AT over time, as observed in Exp. 4 (up to 84% decrease in 4 mo of storage). In Exp. 3, H2 emission was increased by AT and, as expected, the proportion of acetate in the total volatile fatty acids in the rumen was decreased and those of propionate and butyrate were increased by HighAT compared with the control. Compared with the control, HighAT decreased dry matter intake, milk yield, and energy-corrected milk yield in Exp. 3. Milk composition was not affected by treatment, except lactose percentage and yield were decreased by HighAT. Concentrations of iodine and bromide in milk were increased by HighAT compared with the control. Milk CHBr3 concentration and its organoleptic characteristics were not different between control and HighAT. Oregano had no effect on CH4 emission or lactational performance of the cows in Exp. 3. Overall, AT included at 0.50% in the ration of dairy cows can have a large mitigation effect on enteric CH4 emission, but dry matter intake and milk production may also decrease. There was a marked decrease in the CH4 mitigation potential of AT in the second half of Exp. 3, likely resulting from CHBr3 decay over time.

Toxic assessment, growth disrupting and neurotoxic effects of red seaweeds' botanicals against the dengue vector mosquito Aedes aegypti L.

Application of synthetic pesticides over decades to control insects, pests, and disease vectors has resulted in negative impacts on environment and health. The current study assessed the toxicological effects of 12 botanicals obtained from 4 different red seaweeds against the dengue vector mosquito Aedes aegypti L. (Diptera: Culicidae). Four species of red seaweeds, namely Laurencia karachiana, Gracilaria foliifera, Jania rubens, Asparagopsis taxiformis, were collected from Karachi coast and extracted with hexane, dichloromethane and methanol. The efficiency of these extracts was determined by using a dose-response bioassay method against 4th instar larvae of Ae. aegypti. Separate investigations on the toxicity and IGI effects were done. Comparative studies showed that the hexane extracts induced more toxic effects. Based on the LC50 values, obtained after 24 h of treatments, hexane extract of J. rubens (HJ) exhibited toxic effects with LC50 32 µg/mL, (equivalent to GHS category 3), followed by G. foliifera (HG) (LC50 76.8 µg/mL). HJ also showed prominent neurotoxic effects within 1-6 h. Comparatively, higher morphological abnormalities and growth inhibiting (IGI) effects were obtained in the dichloromethane and methanol extracts treated larvae, after 48-96 h, resulting in the formation of immature life forms such as larvi-pupae and pupi-adult. Presumptive growth inhibiting effects were also noted. These included formation of albino and black pupae, deformities in the internal structure of the treated larvae and the chitin synthesis related effects such as 'inhibiting effect on adult emergence'. Finding revealed that red seaweeds, harvested from the Arabian Sea, have potentials to affect Ae. aegypti survival and thus can be utilized as green pesticides.

Effects of local Polynesian plants and algae on growth and expression of two immune-related genes in orbicular batfish (Platax orbicularis).

The emerging orbicular batfish (Platax orbicularis) aquaculture is the most important fish aquaculture industry in French Polynesia. However, bacterial infections are causing severe mortality episodes. Therefore, there is an urgent need to find an effective management solution. Besides the supplying difficulty and high costs of veterinary drugs in French Polynesia, batfish aquaculture takes place close to the coral reef, where use of synthetic persistent drugs should be restricted. Medicinal plants and bioactive algae are emerging as a cheaper and more sustainable alternative to chemical drugs. We have studied the effect of local Polynesian plants and the local opportunistic algae Asparagopsis taxiformis on batfish when orally administered. Weight gain and expression of two immune-related genes (lysozyme g - Lys G and transforming growth factor beta - TGF-ß1) were studied to analyze immunostimulant activity of plants on P. orbicularis. Results showed that several plants increased Lys G and TGF-ß1 expression on orbicular batfish after 2 and 3 weeks of oral administration. A. taxiformis was the plant displaying the most promising results, promoting a weight gain of 24% after 3 weeks of oral administration and significantly increasing the relative amount of both Lys G and TGF-ß1 transcripts in kidney and spleen of P. orbicularis.

Evaluation of Two Species of Macroalgae from Azores Sea as Potential Reducers of Ruminal Methane Production: In Vitro Ruminal Assay.

The utilisation of seaweeds as feed supplements has been investigated for their potential to mitigate enteric methane emissions from ruminants. Enteric methane emissions are the primary source of direct greenhouse gas emissions in livestock and significantly contribute to anthropogenic methane emissions worldwide. The aim of the present study is to evaluate the nutritional role and the in vitro effect on cumulative gas and methane production of Asparagopsis taxiformis (native species) and Asparagopsis armata (invasive species), two species of red algae from the Azorean Sea, as well as the ability to reduce biogas production when incubated with single pasture (Lolium perenne and Trifollium repens) as substrate. Four levels of concentrations marine algae were used (1.25%, 2.25%, 5%, and 10% DM) and added to the substrate to evaluate ruminal fermentation using the in vitro gas production technique. The total amount of gas and methane produced by the treatment incubation was recorded during 72 h of incubation. The results indicate that both algae species under investigation contain relatively high levels of protein (22.69% and 24.23%, respectively, for Asparagopsis taxiformis and Asparagopsis armata) and significant amounts of minerals, namely magnesium (1.15% DM), sodium (8.6% DM), and iron (2851 ppm). Concerning in vitro ruminal fermentation, it was observed that A. taxiformis can reduce enteric methane production by approximately 86%, during the first 24 h when 5% is added. In the same period and at the same concentration, A. armata reduced methane production by 34%. Thus, it can be concluded that Asparagopsis species from the Azorean Sea have high potential as a protein and mineral supplement, in addition to enabling a reduction in methane production from rumen fermentation.

Macroalgal microbiomes unveil a valuable genetic resource for halogen metabolism.

BACKGROUND: Macroalgae, especially reds (Rhodophyta Division) and browns (Phaeophyta Division), are known for producing various halogenated compounds. Yet, the reasons underlying their production and the fate of these metabolites remain largely unknown. Some theories suggest their potential antimicrobial activity and involvement in interactions between macroalgae and prokaryotes. However, detailed investigations are currently missing on how the genetic information of prokaryotic communities associated with macroalgae may influence the fate of organohalogenated molecules. RESULTS: To address this challenge, we created a specialized dataset containing 161 enzymes, each with a complete enzyme commission number, known to be involved in halogen metabolism. This dataset served as a reference to annotate the corresponding genes encoded in both the metagenomic contigs and 98 metagenome-assembled genomes (MAGs) obtained from the microbiome of 2 red (Sphaerococcus coronopifolius and Asparagopsis taxiformis) and 1 brown (Halopteris scoparia) macroalgae. We detected many dehalogenation-related genes, particularly those with hydrolytic functions, suggesting their potential involvement in the degradation of a wide spectrum of halocarbons and haloaromatic molecules, including anthropogenic compounds. We uncovered an array of degradative gene functions within MAGs, spanning various bacterial orders such as Rhodobacterales, Rhizobiales, Caulobacterales, Geminicoccales, Sphingomonadales, Granulosicoccales, Microtrichales, and Pseudomonadales. Less abundant than degradative functions, we also uncovered genes associated with the biosynthesis of halogenated antimicrobial compounds and metabolites. CONCLUSION: The functional data provided here contribute to understanding the still largely unexplored role of unknown prokaryotes. These findings support the hypothesis that macroalgae function as holobionts, where the metabolism of halogenated compounds might play a role in symbiogenesis and act as a possible defense mechanism against environmental chemical stressors. Furthermore, bacterial groups, previously never connected with organohalogen metabolism, e.g., Caulobacterales, Geminicoccales, Granulosicoccales, and Microtrichales, functionally characterized through MAGs reconstruction, revealed a biotechnologically relevant gene content, useful in synthetic biology, and bioprospecting applications. Video Abstract.

The Mediterranean red alga Asparagopsis taxiformis has antifungal activity against Aspergillus species.

The red algae Asparagopsis taxiformis collected from the Straits of Messina (Italy) were screened for antifungal activity against Aspergillus species. EUCAST methodology was applied and extracts showed antifungal activity against A. fumigatus, A. terreus and A. flavus. The lowest minimum inhibitory concentrations observed were <0.15 mg ml(-1) and the highest were >5 mg ml(-1) for Aspergillus spp. tested. Agar diffusion assays confirmed antifungal activity of A. taxiformis extracts in Aspergillus species.

A Proteomic Analysis for the Red Seaweed Asparagopsis taxiformis.

The red seaweed Asparagopsis taxiformis is a promising ruminant feed additive with anti-methanogenic properties that could contribute to global climate change solutions. Genomics has provided a strong foundation for in-depth molecular investigations, including proteomics. Here, we investigated the proteome of A. taxiformis (Lineage 6) in both sporophyte and gametophyte stages, using soluble and insoluble extraction methods. We identified 741 unique non-redundant proteins using a genome-derived database and 2007 using a transcriptome-derived database, which included numerous proteins predicted to be of fungal origin. We further investigated the genome-derived proteins to focus on seaweed-specific proteins. Ontology analysis indicated a relatively large proportion of ion-binding proteins (i.e., iron, zinc, manganese, potassium and copper), which may play a role in seaweed heavy metal tolerance. In addition, we identified 58 stress-related proteins (e.g., heat shock and vanadium-dependent haloperoxidases) and 44 photosynthesis-related proteins (e.g., phycobilisomes, photosystem I, photosystem II and ATPase), which were in general more abundantly identified from female gametophytes. Forty proteins were predicted to be secreted, including ten rhodophyte collagen-alpha-like proteins (RCAPs), which displayed overall high gene expression levels. These findings provide a comprehensive overview of expressed proteins in A. taxiformis, highlighting the potential for targeted protein extraction and functional characterisation for future biodiscovery.

Complex sulfated galactans from hot water extracts of red seaweed Asparagopsis taxiformis comprise carrageenan and agaran structures.

Hot water extraction from the red seaweed Asparagopsis taxiformis yielded three extracts which showed sulfated galactans with a D:L-galactose ratio non consistent with carrageenan or agaran backbones. The major extract was fractionated by cetrimide precipitation and redissolution with increasing sodium chloride concentrations due to their low solubility. Seven fractions were obtained, and studied by methylation analysis, desulfation-methylation, and NMR spectroscopy of the partially hydrolyzed and the native samples. Fractions with the highest yield were those obtained at high concentrations of NaCl. They comprised both agaran and crageenan structures in considerable amounts. The main agaran structures were ß-D-galactose 4-sulfate and ß-D-galactose 2-sulfate units linked to α-L-galactose 2,3-disulfate residues, and ß-D-galactose linked to α-L-galactose 3-sulfate or 6-sulfate, or substituted with single stubs of ß-D-xylose on C3, while the carrageenan structures comprised ß-D-galactose (2-sulfate) linked to α-D-galactose 3-sulfate or 2,3-disulfate, and ß-D-galactose 2,4-disulfate linked to α-D-galactose 2,3-disulfate. Between the less sulfated fractions, the one obtained by solubilization in 0.5 M NaCl was mainly constituted by agarans, which included 3,6-anhydro-α-L-galactose units. Anticoagulant activity was assayed by general coagulation tests (aPTT and TT), showing a moderate action compared with heparin. This is the first detailed study of the sulfated galactans from the order Bonnemaisoniales.

Micro- and Macro-Algae Combination as a Novel Alternative Ruminant Feed with Methane-Mitigation Potential.

This study was conducted to provide alternative high-quality feed and to reduce methane production using a mixture of the minimum effective levels of Euglena gracilis, EG, and Asparagopsis taxiformis, AT. This study was performed as a 24 h in vitro batch culture. Chemical analysis demonstrated that EG is a highly nutritive material with 26.1% protein and 17.7% fat. The results showed that the supplementation of AT as a feed additive at 1 and 2.5% of the diet reduced methane production by 21 and 80%, respectively, while the inclusion of EG in the diet at 10 and 25% through partially replacing the concentrate mixture reduced methane production by 4 and 11%, respectively, with no adverse effects on fermentation parameters. The mixtures of AT 1% with both EG 10% and EG 25% had a greater reductive potential than the individual supplementation of these algae in decreasing methane yield by 29.9% and 40.0%, respectively, without adverse impacts on ruminal fermentation characteristics. These results revealed that the new feed formulation had a synergistic effect in reducing methane emissions. Thus, this approach could provide a new strategy for a sustainable animal production industry.

Optimization of ultrasound-assisted extraction of phenolics from Asparagopsis taxiformis with deep eutectic solvent and their characterization by ultra-high-performance liquid chromatography-mass spectrometry.

Asparagopsis taxiformis is a significant source of phenolics. Owing to the incessant demand of green extraction procedures for phenolics from A. taxiformis, ultrasound-assisted extraction (UAE) using deep eutectic solvents (DESs) was optimized. Among the tested DESs, betaine-levulinic acid afforded the highest total phenolic content (TPC). Moreover, the optimal extraction conditions elucidated from single-factor and response surface methodologies comprised a 52.41°C ultrasonic temperature, 46.48% water content of DES, and 26.99 ml/g liquid-to-solid ratio. The corresponding TPC (56.27 mg GAE/100 g DW) and antioxidant ability fitted the predicted values. UAE afforded superior TPC and antioxidant abilities with DESs than with traditional solvents. Using UHPLC-MS, seven phenolic acids, 18 flavonoids, and two bromophenols were identified and quantified. DES-UAE afforded the highest phenolic compound number (26) and sum of contents. These results disclose the high extraction efficiency of DES-UAE for A. taxiformis phenolics and provide a basis for the higher-value application of this species.

Effects of red macroalgae Asparagopsis taxiformis supplementation on the shelf life of fresh whole muscle beef.

This study was conducted to evaluate the effect of red macroalgae Asparagopsis taxiformis supplementation for cattle on the shelf life of fresh beef steaks (longissimus dorsi). Three treatment groups (seven steers per treatment) included: 1) Control diet, 2) Control diet + 0.25% of macroalgae inclusion (low dose, LD), and 3) Control + 0.5% of macroalgae inclusion (high dose, HD). After the animals were harvested, the strip loins from all animals were collected and aged for 14 days at the meat lab of the University of California, Davis. Then the strip loins were cut into steaks, packaged, and placed on a retail display case for 6 days. During a retail display, instrumental color (L*, a*, and b*) of lean muscle and external fat surfaces were measured every 12 h. Bacterial counts for total aerobic mesophilic bacteria (AMB), aerobic psychrotrophic bacteria (APB), and lactic acid bacteria (LAB) were assessed on days 0, 3, and 6 of retail display. The thiobarbituric acid reactive substances (TBARS) analysis was conducted to measure the lipid oxidation and the pH was also assessed on days 0, 3, and 6. No interactive effect between treatments and time on the shelf life of steaks was observed. The HD red macroalgae supplement decreased (P < 0.05) the lightness (L*) of the surface muscle of the steaks, while the lightness of the external fat was not affected (P < 0.05) by treatments throughout the retail display. The external fat yellowness of the steaks was lower (P < 0.05) in LD and HD treatment groups compared with the control group. An increase (P < 0.05) in counts of AMB, APB, and LAB was observed on the steaks from the steers in the HD treatment group while steaks in Control and LD group had similar bacterial numbers throughout the retail display. The results indicated that the shelf life of steaks from cattle in LD group remained the same as that of the Control group, but the HD of A. taxiformis caused a darker color of steaks with higher microbial counts, which may lead to a shortened shelf life due to undesirable appearance and faster microbial spoilage.

The invasive Asparagopsis taxiformis hosts a low diverse and less trophic structured molluscan assemblage compared with the native Ericaria brachycarpa.

Invasive seaweeds threaten biodiversity and socio-economics values of worldwide marine ecosystems. Understanding to what extent invasive seaweeds can modify local biodiversity is one of the main priorities in conservation ecology. We compared the molluscan assemblage of the invasive Asparagopsis taxiformis with that of the native Ericaria brachycarpa and explore if variation in the molluscan assemblage diversity was related to the substrate attributes (biomass, and thallus, canopy, and interstitial volumes) of the algae. Results showed that A. taxiformis harboured lower diversity and trophic structure of the molluscan assemblage compared to E. brachycarpa. Biomass was the variable that better explained the variation of abundance and number of species as well as the multivariate structure of the molluscan assemblage. Overall, our results suggest that a complete habitat shift from native to invasive species can potentially trigger bottom-up effects in rocky shores habitats, reducing the biodiversity and the services provided by the invaded habitat.

The Mediterranean red alga Asparagopsis: a source of compounds against Leishmania.

Crude extracts and column fractions from the red algae Asparagopsis taxiformis and A. armata from the Strait of Messina (Italy) were screened for the production of antimicrobial compounds. Extracts from both species revealed remarkable antiprotozoal activity against Leishmania, revealing such algae as a great source of natural antiprotozoal products.

Effect of the macroalgae Asparagopsis taxiformis on methane production and rumen microbiome assemblage.

BACKGROUND: Recent studies using batch-fermentation suggest that the red macroalgae Asparagopsis taxiformis has the potential to reduce methane (CH4) production from beef cattle by up to ~ 99% when added to Rhodes grass hay; a common feed in the Australian beef industry. These experiments have shown significant reductions in CH4 without compromising other fermentation parameters (i.e. volatile fatty acid production) with A. taxiformis organic matter (OM) inclusion rates of up to 5%. In the study presented here, A. taxiformis was evaluated for its ability to reduce methane production from dairy cattle fed a mixed ration widely utilized in California, the largest milk producing state in the US. RESULTS: Fermentation in a semi-continuous in-vitro rumen system suggests that A. taxiformis can reduce methane production from enteric fermentation in dairy cattle by 95% when added at a 5% OM inclusion rate without any obvious negative impacts on volatile fatty acid production. High-throughput 16S ribosomal RNA (rRNA) gene amplicon sequencing showed that seaweed amendment effects rumen microbiome consistent with the Anna Karenina hypothesis, with increased ß-diversity, over time scales of approximately 3 days. The relative abundance of methanogens in the fermentation vessels amended with A. taxiformis decreased significantly compared to control vessels, but this reduction in methanogen abundance was only significant when averaged over the course of the experiment. Alternatively, significant reductions of CH4 in the A. taxiformis amended vessels was measured in the early stages of the experiment. This suggests that A. taxiformis has an immediate effect on the metabolic functionality of rumen methanogens whereas its impact on microbiome assemblage, specifically methanogen abundance, is delayed. CONCLUSIONS: The methane reducing effect of A. taxiformis during rumen fermentation makes this macroalgae a promising candidate as a biotic methane mitigation strategy for dairy cattle. But its effect in-vivo (i.e. in dairy cattle) remains to be investigated in animal trials. Furthermore, to obtain a holistic understanding of the biochemistry responsible for the significant reduction of methane, gene expression profiles of the rumen microbiome and the host animal are warranted.

Impoverished mobile epifaunal assemblages associated with the invasive macroalga Asparagopsis taxiformis in the Mediterranean Sea.

There is an increasing concern about the ecosystem consequences of altering macroalgal assemblages. Many macrophytes are foundation species in coastal habitats, supporting much of the biodiversity of these ecosystems by providing essential resources such as food and habitat. The addition of invasive species strongly contributes to habitat modification, but the bottom-up impacts of non-native macroalgae on higher trophic levels remains difficult to predict. The main aim of this study was to evaluate the effects of the invasive macroalga Asparagopsis taxiformis on biodiversity by comparing the mobile macrofauna inhabiting this species to the dominant native species Halopteris scoparia. This is the first comprehensive study of the possible effects of this widespread invasive species on higher trophic levels. A hierarchical sampling design with two different spatial scales was conducted to explore the consistency of the patterns observed. Fifty-nine species belonging to superorder Peracarida were found, accounting 90% of all organisms. A. taxiformis hosted an impoverished epifaunal assemblage in comparison to that associated with the native seaweed, showing significantly lower values of diversity, abundance and number of epifaunal species across study locations. The structure of the associated macrofauna (both in terms of species composition, variability among samples and relative abundance of the species) was also different. Our results highlighted the strong influence of A. taxiformis in the resident community, with differences among the two macroalgae in all the parameters considered. Finally, our results also reflect a biotic homogenization of the epifaunal assemblages associated to A. taxiformis, a scarcely explored consequence of invasive processes in marine environments. Future studies exploring the cascading effects of the observed changes in the epifaunal assemblages would be necessary in order to estimate system responses to macroalgal invasions.

Antioxidant and Antiproliferative Activity of Asparagopsis taxiformis.

BACKGROUND: Asparagopsis taxiformis (Rhodophyta) is a species of red algae belonging to the family Bonnemaisoniaceae. The objective of the present study was to evaluate antioxidant and antiproliferative activity of four fractions (petroleum ether, chloroform, ethyl acetate, and methanol) of A. taxiformis. MATERIALS AND METHODS: The red seaweed, A. taxiformis was collected from Mandapam Coastal Region, Gulf of Mannar, Tamil Nadu. Epiphytes present in algal extracts were cleaned and washed with seawater and fresh water. In vitro antioxidant activity was determined by hydrogen peroxide scavenging, ferric reducing antioxidant power, superoxide radical, metal-chelating activity, and phosphomolybdenum reduction assay. Further, the cytotoxic activity was evaluated using brine shrimp lethality assay. This method is rapid, reliable, inexpensive, and convenient as compared to other cytotoxicity assays. Gallic acid, ethylenediaminetetraacetic acid, ascorbic acid, and quercetin were used as reference antioxidant compounds. RESULTS: Reducing power of chloroform extract increased with increasing concentration of the extract. The radical scavenging activity of extracts was in the following order: ascorbic acid > methanol > chloroform > petroleum ether > ethyl acetate. Highest metal-chelating activity was observed in petroleum ether fractions (63%). Reduction of Mo (VI) to Mo (V) increased in methanol extract (27%) at 100 µg/ml. Moreover, all fractions had an inhibitory effect on the formation of hydroxyl radicals. Results showed that ethyl acetate, methanol, and petroleum ether fractions exhibited potent cytotoxic activity with median lethal concentration values of 84.33, 104.4, and 104.4 µg/ml, respectively. CONCLUSION: Thus, the results showed that red algae possess strong antioxidant and cytotoxic activity that suggests their possible use in the development of pharmaceutical drugs. SUMMARY: Various fractions of red algae Asparagopsis taxiformis was evaluated for in vitro antioxidant and antiproliferative studies. All results indicate potential use of red algae for drug development. Abbreviations Used: Mo: Molybdenum, AlCl3.H2O: Aluminum chloride, NaNO2: Sodium nitrite, NaOH: Sodium hydroxide, H2O2: Hydrogen peroxide, NADH: Nicotinamide adenine dinucleotide, NBT: Nitroblue tetrazolium chloride, PMS: Phenyl methanesulfonate, FeCl2: Ferrous chloride.

Antioxidant properties of the red alga Asparagopsis taxiformis collected on the North West Algerian coast.

The aim of the present study was to evaluate the total phenolic and lipid content, fatty acids profiles and in vitro antioxidant activities of aqueous and solvent extracts of the red seaweed Asparagopsis taxiformis, through six different investigations. The present study demonstrated that phenol contents (mg gallic acid/g dry weight) were highest in the aqueous and methanolic extracts, followed by the ethanolic, hydroethanolic and hydromethanolic extracts. The lowest phenol contents were identified in the three remaining extracts: Butanolic, petroleum ether and acetone extracts. Furthermore, the total lipid content of the algae powder amounted to 2.85% of dry weight. The fatty acid methyl ester profiles analysed by gas-liquid chromatography represented indicated that fatty acids comprised 91.0±0.3% of total algae lipids. The saturated to unsaturated fatty acid contents amounted to 23.2±0.1 and 67.9±0.4% respectively. C13:0 (tridecanoate), C15:0 (pentadecanoate) and C17:0 (heptadecanoate) represented 47.4% of the total saturated fatty acids. Notably, the two most abundant unsaturated fatty acids, C15:1 (pentadecenoate) and C18:2 (octadecadienoate) represented 13.4 and 11.4% respectively, of the total unsaturated fatty acid content. Furthermore, the results of the antioxidant screening performed at 1.0 mg/ml, revealed that aqueous and methanolic extracts exhibited higher inhibition against superoxide and nitric oxide radicals and excellent radical scavenging activity [with half maximal inhibitory concentration (IC50) values 5.1 and 15.0 µg/ml, respectively], demonstrating improved antioxidant behavior when compared with standard ascorbic acid (which has an IC50 value of 3.7 µg/ml). Scavenging activity of the aqueous and methanolic extracts exhibited a strong peroxidation inhibition against linoleic acid emulsion system at a concentration of 300 µg/ml in comparison to the butylated hydroxyltoluene. Although all the studied extracts exhibited ferric reducing power, the aqueous and methanolic extracts had greater hydrogen donating ability. By contrast, hydromethanolic, ethanolic, hydroethanolic, butanolic, acetone and petroleum ether extracts exhibited weak antioxidant behavior. The antioxidant activity of potent seaweed species identified in the current study means that as well as being used as a functional food, they may be developed as novel pharmaceutical compounds and may be used as anti-ageing agents.