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A total of 200 of Crassostrea virginica populations of average weight range from 41.19 ± 3.42 to 47.53 ± 1.06 g were studied to determine the effect of the feed diets of Caulerpa lentillifera and sucrose as growth enhancement. Growth rates increased that range from 56.99 ± 3.16 to 61.56 ± 2.87 g for 90 days period using an artificial water tank system. Previous studies conducted that C. lentillifera contained high protein and were the most abundant component. This seaweed also contained high amounts of minerals and balanced amino acid and notably very rich in iodine. phosphorus, calcium, magnesium and copper that will contribute to oysters growth. Oysters are known to have the ability to uptake dissolved organic matter as nutrients. In the present study, the effects of culture water supplemented with sucrose were tested on oysters. Results revealed that this organic matter promotes growth to the oysters. Sugars will be metabolized into pyruvate through the glycolysis pathway and will result in the supply of energy. Therefore, supplementation of sugar to oysters may have contributed as an energy source together with the lipid and protein content from the algae diet.

Fungal infection mostly caused by marine oomycetes had hindered crustacean production thus searching for natural and safe treatment is currently needed. Thus, this study was conducted to investigate the antimycotic effect of different seaweed extract against marine oomycetes (Lagenidium spp. and Haliphthoros spp) . Two seaweeds species (Eucheuma cottonii and Caulerpa lentillifera) were extracted using ethanol, methanol and water. Each extracts was tested on four fungi strains of marine oomycetes species for minimum inhibitory concentration (MIC) and fungicidal activities. C. lentillifera ethanol extract showed the highest antifungal effect where it can inhibit three from four fungal strains. Meanwhile, E. cottonii ethanol extract has lowest MIC (500 ppm) and inhibit L. thermophilum IPMB 1401 and H. sabahensis IPMB 1402 hyphal growths. Antimycotic effect on zoospores production shows reduction in production after 12 h immersion for three marine oomycetes species. Seaweed extracts toxicity on Artemia sp. showed approximately 5% mortality at 12 h immersion. It is suggested that 12 h immersion of seaweed extract is a suitable treatment for marine oomycetes in aquaculture. This study does not only show potential alternative control method for crab larvae health management, it may also contribute to the sustainable development and food security of aquaculture industry.

Seaweed, Caulerpa sp., is one of the natural materials that contain many kinds of bioactive compounds depending on species and habitat. The aim of this study was to know the effect of sun drying method on the phytochemical content of C. lentillifera from Kei Islands waters, Southeast Moluccas. The study consisted of several steps,C. lentillifera sample collection using sampling purposive method, direct sun drying and indirect wind drying methods, methanol extraction, and phytochemical test using Harborne method. Crude extract of C. lentillifera in indirect wind drying method was higher than that in direct sun drying. Alkaloid, terpenoid and steroid were found in C. lentillifera from Kei Islands on both drying methods. Saponin was only found in indirect wind drying method. Phenolic compounds were not found in C. lentillifera samples from both drying methods

SummaryThe bacterial communities of the sea grape Caulerpa lentillifera were studied during a disease outbreak in Vietnam. The Rhodobacteraceae and Rhodovulum dominated the composition of healthy C. lentillifera. Clear differences between healthy and diseased cases were observed at order, genus and Operational Taxonomic Unit (OTU) level. Bacterial diversity was lower in healthy C. lentillifera, probably because of antimicrobial compounds from the macroalgae and/or from Clostridium, Cutibacterium or Micrococcus bacteria. The likely beneficial role of Bradyrhizobium, Paracoccus and Brevundimonas strains on nutrient cycling and phytohormone production was discussed. The white coloration of diseased C. lentillifera may not only be associated with pathogens but also with an oxidative response. Aquibacter, Winogradskyella and other OTUs of the family Flavobacteriaceae were hypothesized as detrimental bacteria, this family comprises some well-known seaweed pathogens. Moreover, Thalassobius OTU 2935 and 1635 may represent detrimental Rhodobacteraceae. Phycisphaera together with other Planctomycetes and Woeseia were probably saprophytes of C. lentillifera. This study offers pioneering insights on the co-occurrence of C. lentillifera-attached bacteria, potential detrimental or beneficial microbes, and a baseline for understanding the C. lentillifera holobiont. Further metagenomic and biotechnological approaches are needed to confirm functions of some microbes on this macroalgae to enhance food security in the tropics.

Caulerpa lentillifera (sea grape) has been widely used in pharmaceutical industry and health-care products in Thailand. In this study, we attempted to evaluate the toxicity and antioxidant capacity of sea grape extracts in five fractions (ethanol- CLET, hexane- CLHE, ethyl acetate- CLEA, butanol-CLBU, and aqueous-CLAQ). The extracts were evaluated for cytotoxicity by MTT and LDH assays on four cell lines, fibroblast (L929), macrophages (RAW 264.7), hepatocytes (FL83B), and keratinocytes (HaCaT). Genotoxicity was tested by comet assay and micronucleus assay on human lymphoblast cells (TK6). The antioxidant capacity was measured by DPPH and ABTS scavenging assays. Our results demonstrated low cytotoxicity and genotoxicity of CLET, CLBU and CLAQ. When tested by DPPH and ABTS assays, CLET, CLEA, and CLHE showed high antioxidant activity. In conclusion, CLET, CLBU, and CLAQ demonstrated no toxic effects, and CLET, CLEA, and CLHE exhibited high antioxidant capacity. Therefore, our results indicated that CLET, CLEA, and CLHE could be consumed safely at doses lower than 500 and 200 μg/ml for CLHE and CLEA, respectively. Keywords: Anti-oxidation, Caulerpa lentillifera, Cytotoxicity, Genotoxicity

This study aims to determine the physicochemical quality of seagrape (Caulerpa lentillifera) as a freshness label for products cultivated in different seasons. The applied post-harvest storage experiments compared between, within and without seawater that led to oxidative stress conditions. Water content, malondialdehyde (MDA) compound, total phenolic content (TPC), and chlorophyll content were observed at 0, 3, 6, and 9 days of storage. The storage without seawater showed sharper quality reductions by reaching 20–40% of water loss, 70–90% of MDA production, 15–25% of TPC reduction, and 40–60% of total chlorophyll degradation. The storage within seawater showed lower quality reductions due to the specific growth rates still reaching 5–10%. This study found that the greater the physicochemical quality, the slower the decomposition rates of the stored seagrape during storage. Therefore, the seagrapes’ obvious discoloration occurred earlier in winter, followed by summer and spring. Kinetics of chlorophyll degradation on seagrape in different seasons meet different order-reactions during storage. Furthermore, alternating current electric field (ACEF) treatment with 125 kV/m of intensity for 60 min can lower the spring seagrapes’ physicochemical quality by reaching 10–30% of inhibition, resulting in the shelf-life extension for up to 12 days of post-harvest storage.

The integrated aquaculture-seaweed system has been identified as a bio-mitigation strategy to overcome environmental damage, improve the efficiency of nutrient use, maintain good water quality, and ensure the system’s sustainability. This study was conducted to determine the appropriate density of sea grape (Caulerpa lentillifera) in polyculture with whiteleg shrimp (Litopenaeus vannamei) in the same culture tank. Five treatments were randomly designed in triplicate tanks where shrimp was monocultured (without sea grape) as a control treatment and four polyculture treatments with different seaweed density levels (0.5, 1, 1.5, and 2 kg m−3) for 56 days. The results showed that polyculture of shrimp and sea grape significantly reduced the concentrations of total ammonia nitrogen (TAN), nitrite (NO2−), nitrate (NO3−), and phosphate (PO43−) in the rearing tanks and significantly improved (p &lt, 0.05) the growth rate (6.67–6.76% day−1), survival (73.3–78.5%), and production of shrimp (3.44–3.87 kg m−3) compared to monoculture (6.24% day−1, 54.8%, and 2.02 kg m−3, respectively). Applying shrimp and sea grape polyculture at a density of 1 kg m−3 provided a relatively better shrimp performance and feed conversion ratio than other seaweed densities, although not significantly different among polyculture treatments. The findings suggested that sea grape could be used at densities of 0.5–2 kg m−3 in polyculture with whiteleg shrimp, of which 1 kg m−3 resulted in higher production and feed efficiency.

The removal of steroid hormones from the mariculture system using seaweeds (Caulerpa lentillifera, Ulva pertusa, Gracilaria lemaneiformis, and Codium fragile) was investigated. The results illustrated that both 17β-estradiol (E2) and 17α-ethinylestradiol (EE2) could be removed by the seaweeds at different levels, and the Caulerpa lentillifera was the most efficient one. More than 90% of E2 or EE2 at concentration of 10 µg/L was removed by Caulerpa lentillifera within 12 h. Processes including initial quick biosorption, the following slow accumulation, and biodegradation might explain the removal mechanisms of E2/EE2 by Caulerpa lentillifera. E2/EE2 removal was positively related to the nutrient level and the initial concentration of steroid hormone. A significant linear relationship for E2 and EE2 existed between the initial pollutant concentration and the average removal rate. The highest removal kinetic constant (k) value was obtained at 30°C as 0.34 /h for E2 and at 20°C as 0.28 /h for EE2, demonstrating the promising application potential of Caulerpa lentillifera in the water purification of the industrialized mariculture system with relatively high water temperature. Simultaneous and efficient removal of E2 and EE2 by Caulerpa lentillifera was still achieved after 3 cycles in the pilot-scale experiment. The steroid hormones and nutrients in mariculture wastewater could also be simultaneously removed using Caulerpa lentillifera. These findings demonstrated that Caulerpa lentillifera was the promising seaweed for the removal of steroid hormones in mariculture systems.

Herawati VE, Pinandoyo, Ariyati RW, Rismaningsih N, Windarto S, Prayitno SB, Darmanto YS, Radjasa OK. 2021. Effects of Caulerpa lentillifera added into culture media on the growth and nutritional values of Phronima pacifica, a natural fish-feed crustacean Biodiversitas 22: 424-431. Phronima pacifica is a microcrustacean commonly used as natural feed in fish farming as it has high nutrient contents and is suitable based on larval fish mouth gap size and mass cultivation capacity. However, P. pacifica is not produced optimally and its supply is inadequate, so that P. pacifica culturing techniques need to be improved. The culture of P. pacifica is mostly done using sea grape seaweed (Caulerpa lentillifera). The aim of this study was to investigate the optimal stocking density of C. lentillifera added into media culture on the growth and nutritional values of P. pacifica. We applied a complete random design experiment with four treatments of stocking density of C. lentillifera (i.e., 0 g/m2 (Control), 20 g/m2, 40 g/m2, and 60 g/m2) with 3 replicates for each treatment. The following parameters were analyzed, namely specific growth rate (SGR) of C. lentillifera, and the growth, biomass, growth rate, proximate analysis, and amino and fatty acid concentrations of P. pacifica. This study found that the addition of C. lentillifera into culture media significantly enhanced the growth of P. pacifica. The highest effect on the growth was observed in the treatment of 60 g/m2 with C. lentillifera SGR value of 4.76 ± 0.12%/day, and P. pacifica growth rate, population density and biomass of 4.41 ± 0.12 ind/day, 53.81 ± 8.79 ind/L, and 1.14 ± 0.14 g, respectively. Similarly, the highest protein and fat contained in P. pacifica was obtained in the treatment of 60 g/m2 with 60.23% protein and 10.24% fat. Furthermore, the highest fatty acid profile was C20: 5n-3 of 13.23% ± 0.08% and amino acid profile of 45.23% ± 0.01%. The application of the optimal stocking density of C. lentillifera in culture media could increase the growth and nutritional quality of P. pacifica which in turn could enhance aquaculture productivity.

Science & Technology Asia, 26, 2, 128-137

Seaweed was commonly consumed across the worldwide. In Malaysia, Caulerpa lentillifera or known as latok is increasingly popular with as an additional diet by local peoples. The different dietary qualities, minerals and nutrients are of concern to the food industry. This also has potential in pharmaceutical, nutraceutical, bioremediation and agriculture sectors. Despite its complex habitat adaptations, majority of Caulerpa lentillifera in the market has been harvested from the wild, contributing to natural habitat disruptions. Therefore, the purpose of this research was to investigate selected parameters such as salinity, light, and fertilizer of Caulerpa lentillifera under cultivation in Malaysia. The Caulerpa lentillifera showed better growth under salinity above 25 psu and 30 psu, which also provide sufficient light for a period of 10 hours. The optimum growth rate of Caulerpa lentillifera under cultivation conditions can be obtained by adding fertilizer such as conway and von stock media e.g 17.55 % SGR and 13.50 % SGR respectively. This study indicates parameters, including salinity, light and fertilizer that contribute to the growth of Caulerpa lentillifera culture.

Caulerpa lentillifera (sea grapes) is widely consumed in South-East Asia as a low-energy food with high contents of vitamins and minerals. This study investigated dried sea grapes containing 16.6% insoluble fibre commercially produced in Vietnam as an intervention. We hypothesised that insoluble fibre is the primary metabolite that will reverse diet-induced metabolic syndrome. Male Wistar rats (n = 48) were randomly allocated to four groups in a 16 week protocol. Two groups were fed either corn starch (C) or high-carbohydrate, high-fat (H) diets for the full 16 weeks. The other two groups received C and H diets for eight weeks and then received C. lentillifera added to these diets for the final eight weeks (CCL and HCL, respectively). High-carbohydrate, high-fat diet-fed rats developed obesity, hypertension, dyslipidaemia, fatty liver disease and increased left ventricular collagen deposition. C. lentillifera supplementation in HCL rats decreased body weight, systolic blood pressure, plasma concentrations of total cholesterol and non-esterified fatty acids, inflammatory cells in heart and liver, and visceral adiposity. The Firmicutes to Bacteroidetes ratio decreased in the gut microbiota of HCL rats. Therefore, C. lentillifera attenuated cardiovascular and metabolic symptoms of metabolic syndrome in rats, possibly by preventing infiltration of inflammatory cells together with modulating gut microbiota.

Lead is a multi-systemic toxicant when it accumulates and hazardous to health. The aims of this study were to determine the effect of Caulerpa lentillifera aqueous extract on lead accumulation in internal organs and liver functions of lead-intoxicated rats. Twenty four Sprague Dawley rats were randomly assigned to four groups (n=6). Group I was healthy rats treated saline (2 ml/kg of bwt) daily for 28 days. Group II was healthy rats treated with C. lentillifera aqueous extract (500 mg/kg of bwt) daily for 28 days. Group III was intoxicated with lead acetate (20 mg/kg of bwt) daily for 4 days, and then treated with saline (2 ml/kg of bwt) for another 24 days. Group IV was intoxicated with lead acetate (20 mg/kg of bwt) daily for 4 days and then treated with C. lentillifera aqueous extract (500 mg/kg of bwt) for another 24 days. Physical signs of illness were monitored on a daily basis. Lead accumulation in internal organs was measured by atomic absorption spectrophotometer and liver function markers were determined using assay kits. Lead intoxication significantly (p

Three species of Malaysian edible seaweed (Eucheuma denticulatum, Sargassum polycystum and Caulerpa lentillifera) were analyzed for their carotenoid composition using a combination of high-performance thin layer chromatography (HPTLC) and ultra-high-performance liquid chromatography-electrospray ionization-tandem mass spectrometry (UHPLC-ESI-MS/MS), while the antioxidant capacities were determined by 2,2-diphenyl-1-picrylhydrazyl (DPPH) and oxygen radical absorbance capacity (ORAC) assays. The HPTLC analysis exhibited a distinct carotenoid pattern among the three seaweed groups. The UHPLC-ESI-MS/MS analysis showed fucoxanthin as the major carotenoid present in S. polycystum while lutein and zeaxanthin in E. denticulatum. For C. lentillifera, β-carotene and canthaxanthin were the major carotenoids. Some of the carotenoids, such as rubixanthin, dinoxanthin, diatoxanthin and antheraxanthin, were also tentatively detected in E. denticulatum and S. polycystum. For antioxidant activity, S. polycystum (20 %) and E. denticulatum (1128 μmol TE/g) showed the highest activity in the DPPH and ORAC assays, respectively. The findings suggest the three edible varieties of seaweeds may provide a good dietary source with a potential to reduce antioxidative stress.

Background: Liver injury is a common reason for drugs to be withdrawn from the market. Treatment options for common liver disease are limited, and therapy with modern medicines may lack effectiveness. Caulerpa lentillifera may have strong antioxidant systems that protect the plant from oxidative damage caused by the environment.Objectives: The main objective of this study was to evaluate the hepatoprotective effect of the methanolic extract of C. lentillifera against acetaminophen-induced liver toxicity in juvenile zebrafish (Danio rerio). Methods: Juvenile zebrafish (aged 1–3 months) were exposed to 10 mikromolar and 25 mikromolar acetaminophen (N-acetyl-p-aminophenol; APAP) to induce liver damage. C. lentillifera methanolic extracts (0.01 mg/L, 0.02 mg/L and 0.03 mg/L), were concomitantly added to individual tanks containing 0.01 M or 0.025 M APAP. The positive control group was treated with N-acetylcysteine/NAC (0.01 M) and silymarin (0.01 mg/L, 0.02 mg/L and 0.03 mg/L). Hematoxylin and Eosin (H&E) staining revealed the extent of liver injury through the presence of hepatic necrosis, vacuolization, leukocyte infiltration, and ballooning. The antioxidant mechanism of hepatoprotective activity was assessed by a DPPH free radical scavenging assay.Result: C. lentillifera extracts reduced the mortality of juvenile zebrafish when simultaneously exposed to APAP. Upon histopathological examination of the liver tissue of juvenile zebrafish, the group treated with the 0.01 M APAP together with the highest concentration of C. lentillifera extract showed minimal liver injury compared to the groups exposed 0.025 M APAP. However, the DPPH free radical scavenging assay performed using 24–36 mg/mL C. lentillifera extracts showed a minimal effect on the free radical scavenging activity.Conclusion: The histopathological analysis of the liver showed that C. lentillifera extract prevented the progression of liver damage caused by APAP. The results of DPPH free radical scavenging assay indicated that the hepatoprotective activity of C. lentillifera extract might have other antioxidant mechanisms aside from free radical scavenging. In order to effectively assess the improvement in the survival rate of juvenile zebrafish, longer exposure in the treatments is recommendedKeywords: C. lentillifera; juvenile zebrafish; hepatoprotective; drug-induced liver injury (DILI)

Estrada JL, Bautista NS, Dionisio-Sese ML. 2020. Morphological variation of two common sea grapes (Caulerpa lentillifera and Caulerpa racemosa) from selected regions in the Philippines. Biodiversitas 21: 1823-1832. Seagrapes, locally known in the Philippines as “lato” or “ar-arusip”, are economically important macroalgae belonging to the edible species of the genus Caulerpa. This study characterized and compared distinct populations of sea grapes from selected regions in the Philippines and described the influence of physicochemical parameters of seawater on their morphology. Morphometric, cluster and principal component analyses showed that morphological plasticity exists in sea grapes species (Caulerpa lentillifera and Caulerpa racemosa) found in different sites in the Philippines. These are evident in morphometric parameters namely, assimilator height, space between assimilators, ramulus diameter and number of rhizoids on stolon wherein significant differences were found. This evident morphological plasticity was analyzed in relation to physicochemical parameters of the seawater. Assimilator height of C. racemosa is significantly associated and highly influenced by water depth, salinity, temperature and dissolved oxygen whereas for C. lentillifera depth and salinity are the significant influencing factors. Highest assimilator height of sea grapes was found in Coron and Culion in Palawan due to higher salinity and temperature while in Mactan, Cebu, it is primarily attributed to lower depth and higher salinity values.

Herein, water-extracted Caulerpa lentillifera polysaccharides were structurally characterized and their bile acid-binding capacities were investigated. WCLP-25, WCLP-40, WCLP-55, WCLP-70, and WCLP-85 were obtained by graded ethanol precipitation with ethanol concentrations of 25%, 40%, 55%, 70%, and 85%. The total carbohydrate, protein, uronic acid and sulfate contents as well as the monosaccharide composition, molecular weight, and rheological properties were determined. Their infrared spectra, thermogravimetric curves, and scanning electron microscopy (SEM) images were acquired. The hypolipidaemic effects of the WCLPs were assessed with in vitro simulated bile acid-binding capacity experiments. The WCLPs are high-molecular-weight sulfated heteropolysaccharides, and the ethanol concentration significantly influenced the physicochemical properties of the extract. The bile acid-binding capacities of WCLP-55 and WCLP-70 were significantly higher than those of the other tested WCLP samples, which may be due to their higher neutral sugar, uronic acid and sulfate contents or due to their higher viscosities and a larger sheet structure based on SEM. This study will broaden the sources of raw materials for functional foods and provide a reference for the scientific use of C. lentillifera.

Within the siphonous green algal order Bryopsidales, the size and gene arrangement of chloroplast genomes has been examined extensively, while mitochondrial genomes have been mostly overlooked. The recently published mitochondrial genome ofCaulerpa lentilliferais large with expanded noncoding DNA, but it remains unclear if this is characteristic of the entire order. Our study aims to evaluate the evolutionary forces shaping organelle genome dynamics in the Bryopsidales based on theC. lentilliferaandOstreobium quekettiimitochondrial genomes. In this study, the mitochondrial genome ofO. quekettiiwas characterised using a combination of long and short read sequencing, and bioinformatic tools for annotation and sequence analyses. We compared the mitochondrial and chloroplast genomes ofO. quekettiiandC. lentilliferato examine hypotheses related to genome evolution. TheO. quekettiimitochondrial genome is the largest green algal mitochondrial genome sequenced (241,739 bp), considerably larger than its chloroplast genome. As with the mtDNA ofC. lentillifera, most of this excess size is from the expansion of intergenic DNA and proliferation of introns. Inflated mitochondrial genomes in the Bryopsidales suggest effective population size, recombination and/or mutation rate, influenced by nuclear-encoded proteins, differ between the genomes of mitochondria and chloroplasts, reducing the strength of selection to influence evolution of their mitochondrial genomes.

Caulerpa is an unusual algal genus from Caulerpaceae (Chlorophyta, Bryopsidales). Species from this family produce a wide range of metabolites suitable for biotechnology applications. Among these, sulfated polysaccharides (SPs) are often highly desirable for pharmaceutical and nutraceutical applications. Here, we provide a classification of sulfotransferases from Caulerpa, these important enzymes catalyze the nodal step for the biosynthesis of SPs. For this, we performed phylogenetic, genomic, expression analyses and prediction of the protein structure on sulfotransferases from Caulerpa. Sequences, domains and structures of sulfotransferases generally shared common characteristics with other plants and algae. However, we found an extensive duplication of sulfotransferase gene family, which is unique among the green algae. Expression analysis revealed specific transcript abundance in the pinnae and rachis of the alga. The unique genomic features could be utilized for the production of complex SPs, which require multiple and specific sulfation reactions. The expansion of this gene family in Caulerpaceae would have resulted in a number of proteins characterizing the unique SPs found in these algae. We provide a putative biosynthetic pathway of SPs, indicating the unique characteristics of this pathway in Caulerpa species. These data may help in the future selection of Caulerpa species for both commercial applications and genetic studies to improve the synthesis of valuable products from Caulerpa.

Caulerpa lentillifera is an edible and functional seaweed due to its high nutritional compositions and its biological activities. In this study, C . lentillifera was evaluated for its proximate compositions (moisture, ash, protein, lipid and fiber contents) and its biological activities (antimicrobial, anti-oxidant, and toxicity). Moisture content, crude lipid, crude protein, and crude fiber were determined using oven method, soxhlet extraction, semi-micro Kjeldhal, and hydrolysis, respectively. Fresh C . lentillifera of Natuna Island, Indonesia, showed its higher level content of ash, crude lipid, and crude fiber compared to that of fresh C . lentillifera of Penghu, Taiwan. For its biological activity assays, the extracts were prepared from fresh and dry C . lentillifera (FC and DC). Both of the extracts showed the broad spectrum of weak antimicrobial using well-diffusion agar tests and antioxidant activities using a modified linoleic acid emulsion system. The toxicity for both extracts was determined using brine shrimp lethality test. DC extract showed its very low toxicity level and there was no toxicity for FC. Hemolytic activity was determined using red blood assay. Both extracts showed their low hemolytic activities (about 5-13%) for the concentration of 100 and 150 μg/mL, but the activity increased sharply (about 96%) on the concentration of 200 μg/mL. It was concluded that C . lentillifera has a potency as a functional food due to containing secondary metabolites with various biological activities.

In Japan, fresh cultivated green alga, Caulerpa lentillifera (Caulerpaceae, Bryopsidales), are packed into polystyrene containers without seawater. They can stay fresh for 1 week inside the container under room temperature, suggesting a high tolerance to desiccation. This tolerance was evaluated in the present study by employing methods based on pulse amplitude modulation (PAM) fluorometry. The chronological change of the photochemical efficiency of photosystem II (PSII) of C. lentillifera in response to desiccation under conditions experienced during the commercial shipping was examined. Desiccation experiments were carried out for 2, 4, 8, and 12 days at 20 °C. Maximum quantum yields (Fv/Fm) were measured at the initial state, after each desiccation period, and after 10-min and 3, 6, and 24-h re-immersion in seawater. Fv/Fm of samples after a 2-day desiccation remained relatively high (0.78 ± 0.04 standard deviation (SD)), while those after 4- and 8-day desiccation periods decreased and yet were considered active with values greater than 0.40 following rehydration. Samples under the 12-day desiccation had the greatest decline in Fv/Fm, (0.10 ± 0.10), along with 72% critical water loss. Failure of the seaweed to recover from desiccation stress further indicates deactivation of the photosystem, which is perhaps associated with multiple cellular alterations that result at the end in a dysfunctional alga. The packaging system that makes use of polystyrene containers for thermal insulation and an absorbent sheet for moisture control may have provided an optimal environment for the seaweed to extend its “freshness” up to 1 week by maintaining photochemical efficiency.

In this study, four purified fractions (CLGP1, CLGP2, CLGP3 and CLGP4) were prepared from green seaweed Caulerpa lentillifera. They were identified to be a novel kind of xylogalactomanans, differed in molecular weight, monosaccharide composition, and the content of uronic acids and sulfate groups, leading to various ζ-potential, ultrastructure and immunostimulatory activity. Especially, CLGP4 was quite different from the others, as it was found to be a homogeneous heteropolysaccharide composed of Xyl, Man and Gal in a percentage ratio of 1.00:2.15:2.40 with 3877.8kDa. Moreover, CLGP4 contained minor uronic acids (2.37%±0.94%) and the highest sulfate content (21.26%±1.22%). These differences in structural features had an effect on the ζ-potential and ultrastructure of CLGP4, showing rod-, rubble- and ellipsoid-shaped particles with largest negatively charge. In vitro immunostimulatory activity evaluation revealed that all the four fractions significantly stimulated macrophages, but CLGP4 showed more potent immunostimulatory activity due to its stronger function on promoting proliferation of macrophages, enhancing phagocytosis, NO production and acid phosphatase activity in macrophages. Therefore, CLGP4 could be explored as a natural immunomodulator. These results would help a fully exploition of Caulerpa lentillifera polysaccharides recognized as health-improving ingredients in functional foods.

Caulerpa lentillifera (Bryopsidophyceae, Chlorophyta) is an edible seaweed attracting great attention for its expansion of farming scale and increasing consumption in these years. In the present study, a sulfated polysaccharide (CLSP-2) was isolated and separated from C. lentillifera, and its chemical structure was elucidated by a series of chemical and spectroscopic methods. Among these methods, mild acid hydrolysis and photocatalytic degradation were applied to release mono- and oligo-saccharide fragments which were further identified by HPLC-MSn analysis, affording the information of the sugar sequences and the sulfate substitution in CLSP-2. Results indicated that the backbone of CLSP-2 was constructed of →6)-β-Manp-(1→ with sulfated branches at C2, which were comprised of prevalent →3)-β-Galp4S-(1→, →3)-β-Galp2,4S-(1→, and minor Xyl. In addition, the virus neutralization assay revealed that CLSP-2 could effectively protect HeLa cells against SARS-CoV-2 infection with an IC50 of 48.48 μg/mL. Hence, the present study suggests CLSP-2 as a promising agent against SARS-CoV-2.

Background: Caulerpa lentillifera (CL) is a green seaweed, and its edible part represents added value as a functional ingredient. CL was dried and extracted for the determination of its active compounds and the evaluation of its biological activities. The major constituents of CL extract (CLE), including tannic acid, catechin, rutin, and isoquercetin, exhibited beneficial effects, such as antioxidant activity, anti-diabetic activity, immunomodulatory effects, and anti-cancer activities in in vitro and in vivo models. Whether CLE has an anti-inflammatory effect and immune response remains unclear. Methods: This study examined the effect of CLE on the inflammatory status and immune response of lipopolysaccharide (LPS)-stimulated RAW 264.7 cells and the mechanisms involved therein. RAW264.7 cells were treated with different concentrations of CLE (0.1–1000 µg/mL) with or without LPS (1 µg/mL) for 24 h. Expression and production of the inflammatory cytokines, enzymes, and mediators were evaluated. Results: CLE suppressed expression and production of the pro-inflammatory cytokines IL-6 and TNF-α. Moreover, CLE inhibited expression and secretion of the inflammatory enzyme COX-2 and the mediators PGE2 and NO. CLE also reduced DNA damage. Furthermore, CLE stimulated the immune response by modulating the cell cycle regulators p27, p53, cyclin D2, and cyclin E2. Conclusions: CLE inhibits inflammatory responses in LPS-activated macrophages by downregulating inflammatory cytokines and mediators. Furthermore, CLE has an immunomodulatory effect by modulating cell cycle regulators.

Farming the shrimp, fish and another commodity could produce large quantities of waste. Aquaculture waste can be formed as feces, residual feed and dead organism which are accumulated in a cultivation area. Generally, the waste is discharged directly into the water without filtration. Thus, one of the problems that are often faced by farmer is the low quality of pure water due to the high content of nutrients. Moreover, suspended solid pollutant will be formed that lead to eutrophication, oxygen depletion, and precipitation. The aim of this research is to analyze the solid waste pond tiger shrimp as fertilizer for the growth of Caulerpa lentiilifera . Here, the Random Design complete (RAL) and three times in Deuteronomy are used in the experiment. The parameters of one control and three treatment doses are used 0, 2, 4, and 6 g.L -1 respectively. The results showed that solid waste shrimp ponds can be used as fertilizer to meet the needs of Caulerpa lentillifera for growth and the results show the highest value is found in the dose of 6 g.L -1 with NO 3 - (4.58 ppm), NH 4 + (3.34 ppm), PO 4 3- (2.03 ppm) and the value of the rate of growth and the PH are obtained (3.64 g.day -1 ) and (6,4-8), respectively. Keywords: Caulerpa lentillifera, growth rate, Nitrification, solid waste, Tiger Shrimp.

This research is mainly focused on the wastewater treatment using biosorption techniques. The biosorption kinetics of binary sorption of heavy metals by green macro alga Caulerpa lentillifera were studied where such alga was used in the shrimp farm as the nitrogen controller but it grow rapidly so the farmer get rid of its excess amount as waste. The studies of sorption capacity and sorption kinetics of Lead (Pb 2+ ) and Copper (Cu 2+ ) ions in single and binary components were conducted using batch experiments. The new binary sorption kinetic model was developed to predict the experimental results and the calculated outputs from the model were consistent with experimental data.

Background Caulerpa lentillifera is one of the most important economic green macroalgae in the world. Increasing demand for consumption has led to the commercial cultivation of C. lentillifera in Japan and Vietnam in recent decades. Concomitant with the increase of C. lentillifera cultivation is a rise in disease. We hypothesise that epiphytes or other microorganisms outbreak at the C. lentillifera farm may be an important factor contributing to disease in C. lentillifera. The main aims are obtaining differences in the microbial community structure and diversity between healthy and diseased C. lentillifera and key epiphytes and other microorganisms affecting the differences through the results of high-throughput sequencing and bioinformatics analysis in the present study. Results A total of 14,050, 2479, and 941 operational taxonomic units (OTUs) were obtained from all samples using 16S rDNA, 18S rDNA, and internal transcribed spacer (ITS) high-throughput sequencing, respectively. 16S rDNA sequencing and 18S rDNA sequencing showed that microbial community diversity was higher in diseased C. lentillifera than in healthy C. lentillifera. Both PCoA results and UPGMA results indicated that the healthy and diseased algae samples have characteristically different microbial communities. The predominant prokaryotic phyla were Proteobacteria, Planctomycetes, Bacteroidetes, Cyanobacteria, Acidobacteria, Acidobacteria and Parcubacteria in all sequences. Chlorophyta was the most abundant eukaryotic phylum followed by Bacillariophyta based on 18S rDNA sequencing. Ascomycota was the dominant fungal phylum detected in healthy C. lentillifera based on ITS sequencing, whereas fungi was rare in diseased C. lentillifera, suggesting that Ascomycota was probably fungal endosymbiont in healthy C. lentillifera. There was a significantly higher abundance of Bacteroidetes, Cyanobacteria, Bacillariophyta, Ulvales and Tetraselmis in diseased C. lentillifera than in healthy C. lentillifera. Disease outbreaks significantly change carbohydrate metabolism, environmental information processing and genetic information processing of prokaryotic communities in C. lentillifera through predicted functional analyses using the Tax4Fun tool. Conclusions Bacteroidetes, Cyanobacteria, Bacillariophyta, Ulvales and Tetraselmis outbreak at the C. lentillifera farm sites was an important factor contributing to disease in C. lentillifera.

The potential of Caulerpa lentillifera, Gracilaria coronopifolia and Chaetomorpha linum, as biomass feedstock was investigated in this study. It was concluded that seaweed is more suitable for bio-based products synthesis, i.e., bioplastic and bio-lubricants, instead of biofuels due to its relatively low calorific value (~12 MJ/kg). Since seaweed has high moisture content (~80%), hydrothermal liquefaction is recommended, and its efficiency can be further enhanced through microwave technology. Besides, it is found that the thermal degradation of seaweed was best described with the reaction order of 1. The kinetic results also indicated that seaweed consists of lower activation energy (Chaetomorpha linum, as it has no conflict with other interests. Lastly, acetic-acid pre-treatment was suggested to be an optional process in order to increase the algal conversion efficiency as it can reduce up to 25% of ash content.

The process of extracting polysaccharides from the green algae Caulerpa lentillifera was studied by single factor experiments and response surface methodology. Additionally, the immunostimulatory activity of Caulerpa lentillifera polysaccharides (CLP) on RAW264.7 mouse macrophage was evaluated by in vitro cell experiments. The results showed that the optimal extraction conditions consisted of ultrasonification for 30 min, extraction time of 9 h, extraction temperature of 100 °C, and a ratio of water to raw material of 40:1. RAW264.7 macrophage exhibited enhanced phagocytosis with no toxic effects after treatment with CLP. In addition, CLP effectively increased the synthesis and secretion of cytokines (IL-6, TNF-α, IL-1β, and NO), whereby the secretion levels of IL-6, TNF-α, and IL-1β were 1,840.32 ± 21.03 pg/mL (50 μg/mL), 197.17 ± 3.13 ng/mL (50 μg/mL), and 1,178.35 ± 78.82 pg/mL (25 μg/mL), respectively. The polysaccharides contained in Caulerpa lentillifera have potential value for further development due to their immunological activity.

Aquaculture is one of the fastest growing food producing industries in the world. Aquaculture effluent contains high concentrations of inorganic nutrients. Reduction of these inorganic nutrients in aquaculture effluent is crucial for fulfilling the effluent standards or reuse of aquaculture effluent. This study investigated the effective use of green macroalga Caulerpa lentillifera as a bioremediatory species for nutrient removal from aquaculture effluent by conducting an on-station experiment and measurements. The effluent of a fish culture unit was circulated through a macroalgal culture unit every four days for a total of 60 days, allowing 15 circulations. Concentrations of inorganic nutrients (NO2&minus, N, NO3&minus, N, NH3-N, and PO43&minus, ) were measured in the integrated system consisting of a fish unit, settling unit, macroalgal unit and extra tank for water circulation in triplicates. Multiple linear regression analysis revealed that the application of the bioremediation system led to a significant reduction in nutrient concentrations within one day, and slightly further in the following two days. On average over the 15 circulations, the first one day of application decreased the concentrations of NO2&minus, by 0.247 mg/L, 81.6 mg/L, 0.682 mg/L, and 0.352 mg/L, respectively. Furthermore, the C:N ratio of macroalgae decreased during the 60-day application period, providing evidence of the nutrient uptake by macroalgae. Based on the European Union (EU) standard and quality criteria of France and the Joint FAO/WHO Expert Committee (JFWEC), the macroalgae grown in the integrated system were at the safe level for human consumption in terms of contents of Cd, Pb, and As. The results of our study imply that recirculating aquaculture systems utilizing C. lentillifera for biofiltration have the potential for effective treatment of aquaculture effluent integrating fish and macroalgae production.

Determining the optimum levels of macroalgal density and salinity for removing aquaculture effluent has gained increasing research interest in recent years because of the growing concerns over environmental sustainability. Here, we determined the effects of macroalgal density and salinity on the uptake of NO2&minus, NO3&minus, NH3, and PO43&minus, by Caulerpa lentillifera from the effluent of Poecilia latipinna using spectrophotometry. Laboratory experiments were conducted to measure nutrient uptake at five different macroalgal density levels (10, 20, 30, 40, and 50 g/L) and three salinity levels (20, 30, and 40 ppt) with and without aeration. Quadratic regression analysis revealed significant nonlinear and linear effects of macroalgal density on the uptake of NO2&minus, where the maximum uptake was predicted to occur at the macroalgal densities of 31.6, 33.3, 50.0, and 20.0 g/L, respectively. Likewise, the effects of salinity on the uptake of NO2&minus, were significant and nonlinear where the maximum uptake was predicted to occur at the salinity levels of 29.1, 30.7, 29.5, and 29.5 ppt, respectively. The result of the effects of aeration was mixed but somewhat indicated a positive effect on the nutrient uptake within the 24 h period. Our results could help aquaculturists to minimize the excessive nutrients by C. lentillifera from aquaculture effluent while achieving long-term sustainable aquaculture production.

Ultrasonic-assisted extraction based on Response Surface Methodology was applied to isolate a polysaccharide (CLP) from Caulerpa lentillifera obtained in the South China Sea and the highest yield of the crude polysaccharide was 11.8%. CLP consisted of xylose, galactose, glucose and glucuronic acid in a molar ratio of 2.00: 1.00: 0.26: 0.04, with a molecular weight of 115 kDa. The content of uronic acids and sulfate groups in CLP was 14.10% and 27.40%, respectively. CLP presented good radical scavenging activities against O2��-, DPPH radical, while the scavenging activities against OH�� and ABTS radical were not so satisfying.

Nowadays, a circular economy is one of the main strategies for developing a sustainable economy. The objective of this work was to increase the value of Caulerpa lentillifera waste and use it efficiently as a resource for lipid extraction. A crude lipid yield of approximately 27.69% of the dry weight was obtained under the following optimized conditions that consisted of a three-stage extraction with 15 min/stage using a ratio of 1:10 (w/v) at room temperature. The crude extracts contained a total fatty acids (TFA) content of approximately 58.60% of the weight of the extracts, which consisted of 10 types of fatty acids. The proportion of palmitic acid (16:0) as the main component was above 70% of the TFA content. Moreover, it also contained chlorophylls a and b and carotenoids at approximately 1.77, 0.91, and 0.70 µg/mg of the extract, respectively. The total phenolic and flavonoids content, including 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) and 1,1′- diphenyl-2-picrahydrazyl (ABTS and DPPH, respectively) scavenging activities, showed that the lipid extracts had high potential for antioxidant activity. Moreover, the extracts also demonstrated α-glucosidase inhibitory activity with an IC50 value of 8.97 mg/mL. Thus, this extract could be used as an alternative product in a circular economy for preventing diabetes disease.

This study presents the establishment of an integrated multi-trophic aquaculture (IMTA) system in the tropical open waters of southern Cebu, Philippines using a combination of locally available species, namely donkey’s ear abalone (Haliotis asinina) as fed species and seaweeds (Gracilaria heteroclada and Eucheuma denticulatum) as inorganic extractive species. The culture of Caulerpa lentillifera as a biofilter did not work in the open sea cultivation system using baskets. Monthly measurements of shell length, width and body weight of the cultured abalones, together with in situ measurements of physicochemical parameters to assess any changes in water quality, mainly nitrate, nitrite, ammonia and phosphate, were conducted over a year period from February 2013 to January 2014 in three designated stations (Abalone, Seaweed and Control Stations) at three different depths (surface, middle and bottom).Cage culture of abalone side by side with seaweeds in the open sea did not result in any significant water quality disturbance in the area—at least not in the current volume of caged abalones being used. Of the four inorganic compounds monitored in the field, nitrate and ammonia in the Seaweed Station were shown to have relatively lower year-round average values when compared with the Abalone Station, although in the case of nitrate, it was higher in the Control Station compared with the abalone and Seaweed Stations. Although this difference was not significant, it shows the red seaweeds, G. heteroclada and E. denticulatum, to be functioning as a natural filter for these two nutrients. In contrast, nitrite, and phosphate concentrations were not reduced indicating that the seaweeds were not effective biofilter for these two nutrients.The two-month old hatchery-bred donkey’s ear abalones can grow to a size of 53.8 × 28.2 mm (L × W) and body weight of 37.8 g after a period of 12 months. Any expansion of the farm into a much larger commercial-scale farm will have to be complimented with seaweeds stocked around it if only to mitigate possible build-up of excess inorganic wastes—to serve as both a natural filter and as a source of natural feed. The potential use of an organic extractive species has to be further studied under farm condition to achieve its full potential as an IMTA species. Keywords: Donkey, ’s ear abalone, Extractive species, Tropical IMTA, Water quality

Intensive fish aquaculture activities release large amount of nitrogen (NH4-N and NO3-N) to the surrounding waters and lead to water eutrophication. However, integrating a single species of seaweed with the fed fish usually cannot remove both forms of nitrogen efficiently. Polyculture of different seaweeds will be an effective way to reduce the nitrogen load of surrounding waters. To compare the feasibility of using Caulerpa lentillifera and Gracilaria lichenoides as biofilters to treat marine fish aquaculture effluents, nine NH4-N/NO3-N ratios were set up and the nitrogen uptake and growth rate of the two seaweeds were measured. The main results showed that: (1) In the presence of multiple N sources, C. lentillifera selectively takes up NO3-N prior to NH4-N and the NO3-N uptake rate was 7.43–50.43 μmol g−1 (dw) h−1 which was significantly higher than that of G. lichenoides. Gracilaria lichenoides selectively takes up NH4-N prior to NO3-N and the NH4-N uptake rate was 10.27–14.14 μmol g−1 (dw) h−1 which was significantly higher than that of C. lentillifera. (2) The NH4-N/NO3-N ratio had effects on the growth of the two seaweeds: 1/1 and 1/5 of NH4-N/NO3-N were favorable for the growth of C. lentillifera; 1/4 and 1/0.5 of NH4-N/NO3-N were favorable for the growth of G. lichenoides. The results preliminarily indicate that integrated culture of C. lentillifera and G. lichenoides was sufficient for bioremediation of marine fish aquaculture effluents.