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AbstractSelenium (Se) plays a crucial role in human health, influencing conditions such as cancer, diabetes, and neurological disorders. With global population growth and unequal nutrient distribution threatening food security, new approaches are needed to meet the nutritional needs of the world. Se is essential for immune function, metabolism, and antioxidant defense, and in regions suffering from food insecurity and malnutrition, selenium-enriched food could offer an affordable solution. Spirulina platensis, microalgae, can bioaccumulate Se from its environment, enhancing its nutritional value. This study explores how different light spectra (red, white, yellow, and blue LEDs) affect Se bioaccumulation in Spirulina when Na2SeO3 is added to the culture medium in photobioreactors. The results show that red light made the highest Se bioaccumulation (0.118 mg.L−1), followed by white, yellow, and blue light. Se addition also increased cell dry weight by 46%, 33%, 22%, and 60%, respectively, compared to photobioreactors without Se, with biomass productivity highest under red light. Furthermore, Se boosted maximum Chl α concentration, improving photosynthetic efficiency. These findings suggest that optimizing light conditions can significantly enhance the nutritional value of Spirulina, offering a potential solution to global hunger by providing a sustainable, selenium-enriched food source. [ABSTRACT FROM AUTHOR]

Boerhavia diffusa L., (punarnava) is known for its rich, secondary metabolite content and potential pharmacological properties. Boeravinone B, a flavonoid, is a significant plant secondary metabolite found in punarnava, exhibiting various pharmacological properties that translate into anticancer, antioxidant, anti-inflammatory, immunomodulatory and nephroprotective activities. However, the limited production of boeravinone B within the plant poses challenges in meeting market demands. In this study, various biotic components, including filtrates and supernatants of algae, fungi, and bacteria, are employed as elicitors to enhance the production of boeravinone B from the cell suspension cultures of punarnava. Fungal components like yeast extract, Aspergillus niger and Cordyceps militaris , bacteria like Escherichia coli and Bacillus subtilis , as well as Algae such as Valonia utricularis and Spirulina platensis , are utilised to assess their efficiency as elicitors at different day intervals. Results indicate that among all the elicitor treatments, fungal components like yeast extract, C. militaris and A. niger at 100 mg/L, 1 % and 5 % concentrations, administered 6, 6 and 2, days before harvesting, exhibit increased production of boeravinone B by 1.13, 1.14 and 2.63 folds, respectively, when compared to control cultures. Similarly, algae V. utricularis and S. platensis , at 2.5 % concentration and treated before harvesting on Day 6, demonstrate enhanced production of boeravinone B by 1.74 and 4.40 folds compared to control cultures. In addition, the efficiency of various biotic elicitors is examined by quantifying total phenolics and flavonoids in treated cell suspension cultures. These findings have the potential to enhance production strategies and meet the growing demand for this valuable compound with medicinal properties, leveraging easily accessible biotic elicitors. [Display omitted] Biotic components used for elicitation in punarnava cell suspension cultures for the production of Boeravinone B. • Biotic components as elicitors in punarnava cell cultures significantly enhanced boeravinone B production. • Fungal components (yeast extract, Aspergillus niger, Cordyceps militaris) and algae (Valonia utricularis, Spirulina platensis) showed substantial increase in metabolite production. • Optimal concentrations and timing before harvesting were identified, with Aspergillus niger showing the highest enhancement (2.63-fold) and Spirulina platensis (4.40-fold). • Total phenolics and flavonoids quantification supported the efficacy of biotic elicitors, offering promising production strategies. [ABSTRACT FROM AUTHOR]

Natural pigments are desired for use in food products due to the long-term adverse effects of synthetic pigments on consumers, industry, and health. In the search for naturally sourced pigments, researchers explored cyanobacteria and its pigments as one of the potential bioresources. Blue-green algae produce phycobiliproteins such as Phycocyanin, Phycoerythrin, and Allophycocyanin, which can serve as natural pigments. The pigments synthesized and extracted from algal sources have vast commercial value in various industries, including cosmetics, nutraceuticals, pharmaceuticals, etc., apart from their numerous health benefits. Over the course of time, scientific research has led to multiple advancements in the production and extraction strategies of pigments from Spirulina platensis , which has led to the expansion of its applications in various sectors. This review outlines recent advances in the sources and manufacturing of Phycocyanin, with an emphasis on specific extraction and purification procedures, as well as possible applications in diagnostics, foods, cosmetics, and pharmaceutical industries. [Display omitted] • Factors affecting the growth and Stability of Phycocyanin are outlined. • Bioactivities of Phycocyanin extracted from Spirulina is reviewed. • Different extraction methods used for the extraction of PC were documented with their advantages and disadvantages. • Present review also provides an insight on biosynthesis and structure of the Phycocyanin from Phycobiliproteins. [ABSTRACT FROM AUTHOR]

The aim of this study was to develop fruit-based drinks enriched with plant-based proteins [Spirulina platensis biomass (0.25%, w/v) and pea protein isolate (5.83%, w/v)] and probiotics (Lacticaseibacillus casei, Lacticaseibacillus rhamnosus, and Lactiplantibacillus plantarum or their combination) (1%, v/v). The drinks were fermented with probiotics at 37 °C for 24 h before being stored at 4 °C for 90 days. They were then evaluated for microbiological, physicochemical and sensory properties after fermentation and during storage. The probiotic viability in the fermented drinks was between 8.67 and 9.61 log CFU/mL after fermentation. During storage period, the viable probiotic cell counts in the drinks were greater than 7.36 ± 0.09 log CFU/mL, while the pH, titratable acidity, total sugar, protein and total phenolic contents of the probiotic drinks were in the range of 3.44–3.96, 0.69–1.37 g lactic acid/100 mL, 9.14–9.62 g/100 mL, 4.39%–4.70%, and 1053.00–1415 mg GAE/L, respectively. Sensory evaluation demonstrated that the drinks enriched with probiotics were equally or more favourably liked than the control sample during the storage. In general, the sample fermented with L. casei (MC) exhibited the highest probiotic, total phenolic content and antioxidant activity at the end of storage. Conversely, the sample fermented with L. rhamnosus (MR) had a high protein content at the end of storage. Consequently, the enrichment of fruit juices with probiotics and plant proteins offers a new alternative to the functional food market. [ABSTRACT FROM AUTHOR]

The development of efficient photocatalysts is crucial in addressing water pollution concerns, specifically in the removal of organic dyes from wastewater. In this context, the use of silver nanoparticles (Ag NPs) might represent a method to achieve high dye degradation efficiencies. On the other hand, the classical Ag NP production process involves several reactants and operating conditions, which make it poorly sustainable. In the present work, Ag NPs were synthesized according to a new sustainable process involving the use of natural extracts of Spirulina platensis and milder operating conditions. The material was also calcined to determine the influence of organic content on the properties of Ag NPs. The X-ray diffraction (XRD) analysis displayed the AgCl and Ag phases with a crystalline size of 11.79 nm before calcination. After calcination, only the Ag phase was present with an increased crystalline size of 24.60 nm. Fourier Transform Infrared Spectroscopy (FTIR) confirmed the capping role of the metabolites from the extract. Scanning Electron Microscopy (SEM) and Transmission Electron Microscopy (TEM) revealed the spherical or quasi-spherical morphologies with agglomeration due to the calcination. Energy-dispersive X-ray spectroscopy (EDX) and Thermogravimetric (TGA) analyses further confirmed the involvement of metabolites in the synthesis of Ag NPs. The optical changes in the products were observed in a UV-Vis analysis. The Ag NPs were tested for their photocatalytic activity against the laboratory dye brilliant blue r in visible light in various conditions. The highest degradation efficiency of 81.9%, with a kapp value of 0.00595 min−1, was observed in alkaline medium after 90 min of light irradiation. [ABSTRACT FROM AUTHOR]

In this study, phycocyanin obtained from Spirulina platensis extract was used at various concentrations as a natural blue color source in toffee‐type soft candy, and the effect of various storage temperatures (4, 20, and 40°C) on the color stability was investigated. In addition, main quality parameters, such as water activity, texture, and sensory analysis, were determined in the samples stored for 3 months at 20°C, and thus the effect of phycocyanin addition on the product appeal was studied. According to the results, the addition of phycocyanin powder did not significantly affect water activity (p >.05). As expected, L*, a*, and b* values of the samples changed with the phycocyanin, and the ΔE values of the samples stored at 4, 20, and 40°C varied between 0.14–0.44, 0.84–2.76, and 2.63–7.90, respectively, during the entire storage period. The texture analysis outputs revealed that the use of phycocyanin did not cause any change in the textural properties of the samples (p >.05). Considering the sensory analysis, all studied concentrations scored high in terms of color liking, and the use of phycocyanin in toffee‐type soft candy production resulted in a remarkable consumer appeal. Hence, the outcomes of this study show that if temperature control is performed in the production, the organoleptic properties of toffee‐type candy products can be enhanced using phycocyanin and can meet consumer requests and demands. [ABSTRACT FROM AUTHOR]

This study assessed the possible pharmacological effects of Chlorella vulgaris (Cg), Spirulina platensis (St), and silymarin (Sl) against thioacetamide (TA)-induced cardiotoxicity in rats, with a focus on their antioxidant, cardioprotective, and anti-inflammatory properties. The following is the random grouping of sixty male rats into six groups of ten animals each: the control (negative control), TA-intoxicated group (positive control; 300 mg/kg body weight (BW)), Sl + TA group (100 mg Sl/kg BW + TA), St + TA group (400 mg St/kg BW + TA), Cg + TA (400 mg Cg/kg BW + TA), and St + Cg + TA group (400 St + 400 Cg mg/kg BW + TA) were all administered for 30 days. At the start of the study, groups 2 through 6 were administered TA intraperitoneally at a dosage of 300 mg/kg BW for two consecutive days, with a 24 h gap between each dose, to induce cardiac damage. Blood samples were obtained to measure hematological parameters and perform biochemical assays, including lipid profiles and cardiac enzymes. For histopathology and immunohistochemistry determination, tissue samples were acquired. The current findings showed that TA injection caused hematological alterations and cardiac injury, as evidenced by greater serum levels of troponin I, creatine kinase-MB, and total creatine kinase (p < 0.05), as well as significantly elevated serum malondialdehyde and decreased serum total antioxidant capacity (p < 0.05) concentrations. Moreover, an increase in blood low-density lipoprotein and total cholesterol concentration (p < 0.05) was recorded in the TA group. There were alterations in the heart tissue's histological structure of the TA group compared to the control ones. These alterations were characterized by vacuolar degeneration of myocytes, loss of cross striation, coagulative necrosis, and fibrosis of interstitial tissue, which was ameliorated by the supplementation of SI, St, and Cg. The TA-intoxicated group showed weak expression of B-cell lymphoma protein 2 (p < 0.05) and strong immunoreactivity of tumor necrosis factor-α and B-cell lymphoma protein 2-associated X (p < 0.05). However, the groups receiving Sl, St, and Cg experienced the opposite. The administration of Sl, St, Cg, and St + Cg along with TA significantly improved and restored (p < 0.05) erythrogram indices, including RBCs, hemoglobin, total leukocytic count, lymphocytes, and monocyte, to the normal control values. The administration of Sl, St, and Cg alleviated the cardiotoxicity caused by TA via reducing oxidative stress, inflammatory markers, and apoptosis in heart tissue. In summary, the current findings suggest that the treatment with Sl, St, and Cg was beneficial in ameliorating and reducing the cardiotoxicity induced by TA in rats. [ABSTRACT FROM AUTHOR]

Arthospira platensis and Spirulina platensis microalgae are a rich source of pro-health metabolites (% d.m.): proteins (50.0–71.3/46.0–63.0), carbohydrates (16.0–20.0/12.0–17.0), fats (0.9–14.2/6.4–14.3), polyphenolic compounds and phenols (7.3–33.2/7.8–44.5 and 4.2/0.3 mg GAE/g), and flavonoids (1.9/0.2 QUE/g) used in pharmaceutical and cosmetic formulations. This review summarises the research on the chemical profile, therapeutic effects in dermatological problems, application of Arthrospira and Spirulina microalgae, and contraindications to their use. The pro-health properties of these microalgae were analysed based on the relevant literature from 2019 to 2024. The antiviral mechanism of microalgal activity involves the inhibition of viral replication and enhancement of immunity. The anti-acne activity is attributed to alkaloids, alkanes, phenols, alkenes, phycocyanins, phthalates, tannins, carboxylic and phthalic acids, saponins, and steroids. The antibacterial activity generally depends on the components and structure of the bacterial cell wall. Their healing effect results from the inhibition of inflammatory and apoptotic processes, reduction of pro-inflammatory cytokines, stimulation of angiogenesis, and proliferation of fibroblasts and keratinocytes. The photoprotective action is regulated by amino acids, phlorotannins, carotenoids, mycosporins, and polyphenols inhibiting the production of tyrosinase, pro-inflammatory cytokines, and free oxygen radicals in fibroblasts and the stimulation of collagen production. Microalgae are promising molecular ingredients in innovative formulations of parapharmaceuticals and cosmetics used in the prophylaxis and therapy of dermatological problems. This review shows the application of spirulina-based commercial skin-care products as well as the safety and contraindications of spirulina use. Furthermore, the main directions for future studies of the pro-health suitability of microalgae exerting multidirectional effects on human skin are presented. [ABSTRACT FROM AUTHOR]

The aim of this study was to investigate the potential of upcycling Expanded Polystyrene (EPS) waste collected from food packaging into a membrane for microalgae harvesting, in which membrane filtration often challenges fouling and pore blocking. The target species is Spirulina platensis, with Chlorella vulgaris as a comparison agent. The membrane was fabricated from used Styrofoam, which typically ends up as single-use food packaging waste. In this study, PVP was used as an additive at varying concentrations, from 2 wt.% to 8 wt.%. The experimental results indicated that despite varying PVP concentrations, all EPS waste membranes exhibited near-complete recovery of Spirulina platensis biomass extraction. Despite the similar harvesting efficiency, EPS/PVP-8 exhibited the largest flux of 970.5 LMH/Bar, which is twice the value of the pristine EPS waste membrane. All membranes were hydrophilic; however, hydrophobicity increased with PVP concentration. SEM micrographs revealed that PVP enlarged the membrane surface pores and improved connectivity within the membrane's structure, ensuring efficient flow. The EPS waste membrane offers promising insights for sustainable materials and wastewater treatment. The upcycling of EPS waste into flat sheet membranes not only addresses the problem of Styrofoam waste accumulation but also paves the way to transform waste into valuable products. [ABSTRACT FROM AUTHOR]

• Assessing the effect of prebiotic on Lactobacillus casei. • Combining Spirulina platensis and L. casei. • Using molecular studies to confirm the mixture efficacy. • The present work elucidated the magnificent role of Spirulina as an immunomodulatory agent through being a prebiotic agent to mediate the antibacterial responses of L. casei. Microalgae are diminutive unicellular magnificent creatures that exist in the microscopic form. Within this classification, two notable subtypes, namely Spirulina platensis and Chlorella vulgaris , have garnered significant attention. S. platensis is mostly known for its therapeutic benefits due to proteins, vitamins, essential amino acids, minerals, and essential fatty acids abundance. The subsequent work aimed to investigate the prebiotic effect of S. platensis enhancing Lactobacillus casei microbiome growth rate and antibacterial effect. Different concentrations of S. platensis were prepared (0, 0.5, 5, and 50 mg/mL) then co-cultured with L. casei. Live probiotic enumeration, molecular characterization and scanning electron microscopy studies were performed. Data revealed that 5 mg/mL of S. platensis significantly enhanced the probiotic live count and consequently the antibacterial activity. The antibacterial and antibiofilm effects of L. casei / S. platensis mixture showed complete inhibition to the growth of bacterial cells after 4- and 8-h incubation with Staphylococcus epidermidis 12228 and Escherichia coli 25922, respectively. Moreover, minimal biofilm eradication concentration (MBEC) value of 16 µg/mL was recorded for L. casei / S. platensis mixture [ABSTRACT FROM AUTHOR]

ackground. Some studies have shown that Spirulina has biological properties exerting beneficial effects on human health. This study aimed to evaluate the effects of Spirulina supplementation on serum glucose, lipid parameters, and oxidative stress markers in male rats with streptozotocin/nicotinamide–induced T2DM. Methods. In this experimental study, rats with diabetes were divided into six groups: healthy control, diabetic control, diabetic-metformin, diabetic-Spirulina 100 mg/dL, diabetic-Spirulina 200 mg/dL, and healthy control-Spirulina 200 mg/d. At the end of the study (28 days), the blood samples were collected, and the fasting blood glucose (FBG), triglycerides (TG), total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), malondialdehyde (MDA), and total antioxidant capacity (TAC) were determined. Results. Significant reductions (P<0.001) were found in serum level of FBG in Metformin and Spirulina 100 mg/kg compared with diabetic control group. The groups were different regarding the serum levels of HDL-C post-intervention. ANOVA analysis also showed significant differences between the Spirulina 100 mg/kg or Metformin group and diabetic control group regarding the serum level of MDA (P<0.05). Conclusion. Spirulina at a dose of 100 mg/kg may have contributed to controlling the fasting blood glucose and oxidative stress. Practical Implications . Our study results suggested that Spirulina, as a promising agent, at lower doses may have been considered as a functional food for the management of diabetes mellitus. [ABSTRACT FROM AUTHOR]

Phycocyanin was extracted from Spirulina platensis using conventional extraction (CE), direct ultrasonic-assisted extraction (direct UAE), indirect ultrasonic-assisted extraction (indirect UAE), and microwave-assisted extraction (MAE) methods at different temperatures, extraction intervals, stirring rate, and power intensities while maintaining the same algae to solvent ratio (1:15 w/v). The optimization of the extraction parameters indicated that the direct UAE yielded the highest phycocyanin concentration (29.31 ± 0.33 mg/mL) and antioxidant activity (23.6 ± 0.56 mg TE/g algae), while MAE achieved the highest purity (Rp = 0.5 ± 0.002). Based on the RP value, phycocyanin extract obtained by MAE (1:15 w/v algae to solvent ratio, 40 min, 40 °C, and 900 rpm) was selected as active compound in an alginate-based hydrogel formulation designed as potential wound dressings. Phycocyanin extracts and loaded hydrogels were characterized by FT-IR analysis. SEM analysis confirmed a porous structure for both blank and phycocyanin loaded hydrogels, while the mechanical properties remained approximately unchanged in the presence of phycocyanin. Phycocyanin release kinetics was investigated at two pH values using Zero-order, First-order, Higuchi, and Korsmeyer-Peppas kinetics models. The Higuchi model best fitted the experimental results. The R2 value at higher pH was nearly 1, indicating a superior fit compared with lower pH values. [ABSTRACT FROM AUTHOR]

Simple Summary: Microalgae such as Spirulina platensis and essential oils derived from Citrus limon have potential growth-promoting, antimicrobial, antioxidant, and immunostimulant effects for several fish species. This manuscript spotlights Spirulina platensis and Citrus limon essential oil and their roles in improving the performance and health of Nile tilapia. Our results indicate that Spirulina platensis and/or lemon essential oil can be used as natural feed additives during aquafeed formulation to improve fish welfare through nutritional management. The dietary presence of feed additives is crucial for boosting fish growth and immunity. Accordingly, this feeding trial aimed to investigate the effects of the separate and concurrent dietary supplementation of Spirulina platensis (SP) and bitter lemon (Citrus limon) peel essential oil (LEO) on the growth, immunity, antioxidant capacity, and intestinal health of Nile tilapia (Oreochromis niloticus). Four groups of male Nile tilapia were employed. The first group (control) was given the basal diet, while the second and third groups received the basal diet supplemented with LEO extract (1%) and SP (1 g/kg diet), respectively. The fourth group received the basal diet supplemented with a mix of LEO (1%) and SP at 1 g/kg. After two months of feeding, using LEO or/and SP improved the overall growth and immunological parameters, with their combination yielding the best outcomes. The supplementation of LEO or/and SP improved the Nile tilapia's growth metrics and transcriptomic levels of growth-regulating genes such as (oligo-peptide transporter 1 (Pep1), growth hormone receptors 1 (GHR1), and insulin-like growth factor (IGF1). The improved growth performance was linked to significant increases in the expression levels of mucin and fat metabolism-related genes. Moreover, fish supplemented with LEO, SP, or their combination showed enhanced non-specific immunological measures, including phagocytic and lysozyme activities and the mRNA copies of its regulating genes. Additionally, remarkable increases in the antioxidant enzyme activities and the mRNA levels of their related genes were detected. The complement (C3) gene's transcriptomic level was also significantly increased. Furthermore, the dietary supplementation of LEO, SP, or their combination improved the histological structures of the spleen, hepatopancreas, and intestine. The enhanced effects of LEO, SP, or their combination on fish immunity and growth are suggested to be due to their contents of bioactive compounds with anti-inflammatory, antioxidant, and antimicrobial properties. Thus, using the LOE and SP blends as feed additives is recommended for better growth and immunity of Nile tilapia. [ABSTRACT FROM AUTHOR]

Milk is a suitable medium for the growth of many microbes, in particular, fungi and yeasts that change the physical and chemical properties of milk and in the final dairy products. In the current study, the target was to apply natural components to control growth of Candida albicans in pasteurized milk. Chitosan (CS), chitosan nanoparticles (CSN), and Spirulina platensis (SP) and its extract (SPE) were used. The MIC (12.5 to 100 mg/ml) of the natural components against Candida albicans as a model for fungi family was determined their effect in broth and in pasteurized milk. CS and CSN were the most effective natural component inhibit the growth of C. albicans strain with concentration 25 mg/ml followed by SPE with inhibitory activity against C. albicans at concentration 100 mg/ml while, Spirulina platensis (SP) showed the least inhibitory activity with the same concentration. By application in pasteurized milk CSN showed the best candidacidal effect with inhibitory 1.2 log10cfu/ml after 15 days at refrigerated storage as adding CSN extended shelf life of pasteurized milk. In conclusion, we recommend adding chitosan nanoparticles in milk at concentration 25mg/ml to limit the fungal growth. [ABSTRACT FROM AUTHOR]

There is a lack of dietary fiber and some essential amino acids in instant noodles. Enriching this popular food with protein‐ and fiber‐rich sources is important to improve the nutritional quality of noodles. The current study is aimed at enriching instant noodles by substituting wheat flour with lentil flour (7%–35%: L7%, L21%, and L35%), soy‐protein isolate (1%–5%: S1%, S3% and S5%), egg‐white protein (1–5: E1%, E3%, and E5%) and Spirulina platensis (1%–5%: S.p1%, S.p3% and S.p5%). The physicochemical, microbial, textural, and sensorial properties of noodle samples were investigated. The spatial relationship between parameters was also evaluated based on principal component analysis (PCA) to select the suitable noodle formulation. The highest protein (17.06%) and lowest carbohydrate contents (76.44%) were for L35%. The lowest (1.69%) and highest fat contents (2.04%) were for S5% and S.p5%m, respectively (p > 0.05). Energy values varied from 394.83 (E5%) to 396.37 kcal/100 g (S.p5%). There was no significant difference between the microbial quality of noodle samples (p > 0.05). The hardness of noodles with 1%–5% soy protein, 21% or more lentil flour, and 3% or more egg‐white/Spirulina was higher than the control/unenriched group (p < 0.05). The color difference of E1 and E3% with the control sample was not obvious (ΔE∗ < 3). Although all the ingredients improved the nutritional value of the noodles, the overall acceptance of samples with 3% or more of Spirulina was lower than the acceptable limit (a score of 3). According to PCA, when the nutritional value and sensory acceptance are important, the L35% may be a better choice. E1%, E3%, S1%, S5%, and L7% noodles received almost the same sensory score as the control sample, while they had more nutritional values. A combination of animal, plant, and microalgae protein sources may provide a noodle with high nutritional value, containing a wide range of essential amino acids and bioactive compounds. More research is needed to optimize such a formulation. [ABSTRACT FROM AUTHOR]

The failure of mass production of Spirulina plateaus can be attributed to an imbalance of nutrients (C:N) and an increase in accumulated sodium ions, coupled with the traditional harvesting process. The current study aims at the recovery of stressed and red cultures of Spirulina platensis as well as enhanced phycocyanin accumulation. The stressed Spirulina platensis cultures were obtained from a local Egyptian Spirulina production farms, which were further subjected to water analyses after removing the Spirulina biomass. Optimization was performed within 300-ml water path photobioreactor. Spirulina platensis samples were incubated with Zarrouk medium comparing with those modified using ammonium bicarbonate or ammonium acetate instead of sodium bicarbonate. Continuous batching was performed every 12 days during three sequenced batches. Growth measurements (dry weight and pigments) were performed along the incubation time. It was found that carbon content of the growth medium seems to be more effective in Spirulina growth and biomass characteristics. Under different carbon sources, acetate resulted in the maximum dry weight of 1.48 g·l−1 and recovery percentage of 463.3%. Such effect was extended along the different incubation batches. Various carbon concentrations revealed that moderate concentration of carbon in the form of acetate (0.699 g·l−1) leads to the maximum growth under the same nitrogen content. A similar trend was observed with chlorophyll and phycocyanin accumulation, while carotenoids showed the opposite manner. [ABSTRACT FROM AUTHOR]

The development of serum-free media (SFM) is critical to advance cell culture techniques used in viral vaccine production and address the ethical concerns and contamination risks associated with fetal bovine serum (FBS). This study evaluated the effects of marine microalgal extracts and growth factor cocktails on the activity of Madin-Darby canine kidney (MDCK) and Vero cells. Five marine microalgal species were used: Spirulina platensis (SP), Dunaliella salina (DS), Haematococcus pluvialis (HP), Nannochloropsis salina (NS), and Tetraselmis sp. (TS). DS and SP extracts significantly increased the proliferation rate of both MDCK and Vero cells. DS had a proliferation rate of 149.56% and 195.50% in MDCK and Vero cells, respectively, compared with that in serum-free medium (SFM). Notably, DS and SP extracts significantly increased superoxide dismutase (SOD) activity, which was 118.61% in MDCK cells and 130.08% in Vero cells for DS, and 108.72% in MDCK cells and 125.63% in Vero cells for SP, indicating a reduction in intracellular oxidative stress. Marine microalgal extracts, especially DS and SP, are feasible alternatives to FBS in cell culture as they promote cell proliferation, ensure safety, and supply essential nutrients while reducing oxidative stress. [ABSTRACT FROM AUTHOR]

Background: Methotrexate (MTX), a drug utilized in cancer and rheumatoid arthritis treatment, is associated with acute and chronic neurodegenerative alterations. Spirulina platensis (SP) has several important phytochemical substances that act as free radical scavengers or natural antioxidants. The current study investigated the possible effects of the blue-green alga Spirulina platensis on cerebellar damage in male rats exposed to methotrexate. Forty (40) adult male albino rats were randomly divided into 4 groups (n = 10) and treated for one week: GI, the control group; GII was orally given 1000 mg SP/kg/daily, GIII was given a single intraperitoneal injection of MTX 75 mg/kg at the first day, and continued under the normal condition without other treatment till the end of the experiment, and GIV received both SP and MTX together with the same previous doses and duration. Neurobehavioral, histopathological, histochemical, immunohistochemical, ultrastructural, molecular, and biochemical data were recorded. Results: MTX caused severe cerebellar degeneration in 3 cortical layers, especially the Purkinje layer. The Purkinje layer displayed a disrupted monolayer arrangement with pyknotic nuclei, a significant decrease in cell number, and shrunken cells surrounded by empty spaces. The molecular and granular layers are degenerated with elevated immunoreactions and gene expression of the glial fibrillary acidic protein (GFAP), ionized calcium-binding adapter molecule 1 (Iba-1), and neurofilament light chain antibody (NFL). Moreover, MTX significantly increased malondialdehyde (MDA) and myeloperoxidase (MPO) while decreasing the levels of reduced glutathione (GSH), serotonin, superoxide dismutase (SOD), acetylcholinesterase (ACHE), norepinephrine, and dopamine. These insults were noticeably mitigated by concomitant treatment with spirulina. Conclusion: Spirulina improves neurological function by modulating the cerebellar damage elicited by MTX. This improvement may be attributed to the anti-inflammatory and antioxidant properties of spirulina. [ABSTRACT FROM AUTHOR]

Microalgae-based wastewater treatment offers an eco-friendly opportunity for simultaneous nutrient recovery and biomass generation, aligning with the circular bioeconomy concept. This approach aims to utilize the nutrients of potato industry wastewater (PIW) for algal growth while mitigating the environmental impact of this industrial byproduct. This study focused on cultivating three cyanobacterial strains, Anabaena oryzae, Nostoc muscorum, and Spirulina platensis, in PIW and synthetic media for 30 days to assess feasibility. Growth performance was monitored by measuring chlorophyll content, dry weight (DW), optical density (OD), and pH at 3-day intervals. The high-performing cyanobacterial biomass from the laboratory findings was formulated into a biofertilizer, which was then evaluated in a controlled greenhouse experiment on celery and lettuce plants. The biofertilizer replaced conventional NPK mineral fertilizers at different levels (25%, 50%, and 75%), while a control group received 100% chemical fertilizer. The results showed favourable growth of all three cyanobacteria strains and their mixture in PIW throughout the experiment. The mixed cyanobacteria followed by Spirulina platensis exhibited the highest growth rates, achieving chlorophyll contents of 3.75 and 2.30 µg·mL−1, DWs of 1.79 g·L−1 and 1.63 g·L−1, and ODs of 0.41 and 0.38, respectively, surpassing the other treatments. The formulated biofertilizers, Spi-PIW (Spirulina platensis + potato industry wastewater) and Cyano-PIW (mixed culture+ potato industry wastewater), significantly enhanced plant height, root and stem lengths, and the number of leaves per plant in celery and lettuce compared to the control group. These biofertilizer treatments also improved chlorophyll contents, as well as macro- and micronutrient levels, in the two crops. Additionally, the application of these biofertilizers improved certain sandy soil properties, i.e., pH, total organic matter, total nitrogen, phosphorus, and potassium. In conclusion, utilizing PIW as a substrate for cultivating cyanobacteria strains and producing high-quality liquid bio-organic fertilizers holds potential for reducing recommended NPK fertilizer doses by 25–50% in celery and lettuce growth, providing an environmentally friendly approach. [ABSTRACT FROM AUTHOR]

Today, the use of nutrients derived from natural bioactive compounds application in the food, pharmaceutical, and cosmetic industries is on the increase. This paper aimed to evaluate the amino acids profile (essential and non-essential) and pigments composition (chlorophyll a, carotenoids, and phycocyanin) of Arthrospira platensis (a blue-green microalga) cultivation in a flat-plate photobioreactor under various types of light-emitting diodes (red: 620–680 nm, white: 380–780 nm, yellow: 570–600nm, blue: 445–480 nm). The maximum biomass concentration (604.96 mg L−1) occurred when the red LED was applied for cultivation, and the minimum biomass concentration (279.39 mg L−1) was obtained under blue LED. The sequence of pigments and amino acids concentrations (mg L−1culture volume) was approximately in accordance with the biomass productivity. It means the red light produces the maximum concentration of pigments (chlorophyll a: 5.42, carotenoids: 2.92, phycocyanin: 67.54 mg L−1) and amino acids (essential amino acids: 110.47, nonessential amino acids: 179.10 mg L−1). Nevertheless, when these values were measured in mg per g of dry weight, the utmost contents were observed in microalgal products cultivated under blue LED. These consequences are due to the highest cell productivity and the most extended length of cells that occurred under red and blue LEDs, respectively. Highlights: Spectral light quality is an effective parameter on Spirulina platensis growth. Red light produces the highest biomass productivity. Pigment and amino acids concentrations have conformity with biomass productivity. Blue light causes the cells to be longer. Longer cells have more time to synthesize the intracellular compounds. [ABSTRACT FROM AUTHOR]

BACKGROUND: The efficient separation and purification of proteins like C‐phycocyanin (C‐PC) from Spirulina platensis are essential for their commercialization, yet they remain challenging. This study investigated three chromatographic methods for C‐PC purification: weak anion exchange chromatography (DEAE), strong anion exchange chromatography (Q Sepharose), and hydrophobic interaction chromatography (HIC). RESULTS: Weak anion exchange chromatography achieved a recovery of 36.80 mg unit (57.08%) with a purity of 3.23, outperforming Q Sepharose (yield: 23.21 mg unit means that 46.33%, purity: 2.76) and HIC (yield: 22.95 mg unit means that 17.57%, purity: 3.02). The purified C‐PC consisted of α and β subunits with molecular masses of 16 kDa and 17 kDa, respectively. Further assessment revealed its antioxidant capacity through a 2,2'‐Azino‐bis(3‐ethylbenzothiazoline‐6‐sulfonic acid) (ABTS) assay. The stability of C‐phycocyanin was tested at different pH levels and temperatures. Maximum stability was observed at pH 7, and pH 4 showed the lowest stability. Glutaraldehyde‐treated C‐PC (GC‐PC) demonstrated gradual degradation up to 50 °C, retaining 73.25% after 30 min. Notably, GC‐PC exhibited stability even at higher temperatures, with degradation rates of 57.32% at 70 °C and 50.96% at 80 °C. CONCLUSION: Weak anion exchange chromatography proved superior for C‐PC purification, offering higher yields and purity than Q Sepharose and HIC. The purified C‐PC showed promising antioxidant capacity and stability, particularly GC‐PC, which exhibited resistance to degradation, even at elevated temperatures. These findings underscore the potential of C‐PC as a valuable compound for various applications, with DEAE chromatography being an efficient method for its production and commercialization. © 2024 Society of Chemical Industry. [ABSTRACT FROM AUTHOR]

Soil salinity, which affects plant photosynthesis mechanisms, significantly limits plant productivity. Soil microorganisms, including cyanobacteria, can synthesize various exometabolites that contribute to plant growth and development in several ways. These microorganisms can increase plant tolerance to salt stress by secreting various phytoprotectants; therefore, it is highly relevant to study soil microorganisms adapted to high salinity and investigate their potential to increase plant resistance to salt stress. This study evaluated the antioxidant activity of four cyanobacterial strains: Spirulina platensis Calu-532, Nostoc sp. J-14, Trichormus variabilis K-31, and Oscillatoria brevis SH-12. Among these, Nostoc sp. J-14 presented the highest antioxidant activity. Their growth-stimulating effects under saline conditions were also assessed under laboratory conditions. These results indicate that Nostoc sp. J-14 and T. variabilis K-31 show significant promise in mitigating the harmful effects of salinity on plant size and weight. Both strains notably enhanced the growth of Oryza sativa plants under saline conditions, suggesting their potential as biostimulants to improve crop productivity in saline environments. This research underscores the importance of understanding the mechanisms by which cyanobacteria increase plant tolerance to salt stress, paving the way for sustainable agricultural practices in saline areas. [ABSTRACT FROM AUTHOR]

This interventional clinical trial aimed to assess the potential impact of Spirulina platensis supplementation on pregnant and lactating ewes living in a Moroccan endemic fluorosis area. Forty-eight ewes were divided into four equal groups: Groups I and II served as controls belonging respectively to fluorosis-free and endemic fluorosis areas, Groups III and IV received respectively 250 and 500 mg*kg−1 BW/day of S. platensis , during late pregnancy and early lactation. The results revealed that ewes reared in fluorosis-free areas exhibited significantly lower plasma fluoride and significantly higher haemoglobin levels compared to endemic fluorosis areas (P < 0.0001). However, supplementation with 500 mg*kg−1 BW*day−1 of S. platensis significantly improved these two parameters compared to Group II (P < 0.0001). Ewes in the endemic area also displayed increased oxidative stress (P < 0.05), characterized by decreased ascorbic acid levels and catalase activity, as well as elevated levels of reduced glutathione and malondialdehyde. Supplementation with 500 mg*kg−1 BW*day−1 of S. platensis enhanced the antioxidant status (P < 0.05) by increasing ascorbic acid levels and catalase activity and decreasing levels of reduced glutathione and malondialdehyde. Moreover, this dose yielded similar average daily gains compared to lambs of ewes living in fluorosis-free area. In conclusion, S. platensis may serve as a promising solution for addressing endemic fluorosis in pregnant and lactating ewes. [ABSTRACT FROM AUTHOR]

Biological nanoparticles can fight drug-resistant viruses and bacteria as therapeutic antiviral and antibacterial agents, assisting in the development of antiviral and antibacterial drugs. The blue-green algae Spirulina platensis was used in a previous study, mediated by Ag-NPs and CuO-NPs. In the present study, both kinds of nanoparticles (NPs) were investigated against Adeno virus7 (AdV-7) which has been indicated by its reduction activity of cytopathic effect (CPE). Using Vero cells, cytotoxicity was assessed and quantified using the MTT assay. Results showed a 90% reduction in CPE of Ad-7 applying Ag -NPs and CuO -NPs at 2.44 μ g/mL, with a higher reduction rate (45.7%) with Ag-NPs than that of CuO-NPs (37.1%). The results demonstrated the efficacy of green nanotechnology applications utilizing both Ag-NPs and CuO-NPs as reducing and inhibitory agents for AdV -7 replication. Moreover, Ag-NPs and CuO-NPs are found to have effective antibacterial properties, they exhibited significant inhibitory activity against certain Gram-negative and Gram-positive bacteria. When compared to the control, Ag-NPs were more effective than CuO-NPs, which had high significant MIC results against, Pseudomonas aeruginosa ATCC 9027, Escherichia coli ATCC 8739, and Salmonella typhi ATCC 12023 (31.25 μ g/ml), while Staphylococcus aureus ATCC 6598 (15.62 μ g/ml). Current results proved the efficiency of green nanotechnology application with both Ag-NPs and CuO-NPs, as reducing and inhibitory agents for the Ad-7 replication and antibacterial activities. [ABSTRACT FROM AUTHOR]

The development of nanoparticles derived from noble metals has garnered significant attention in recent years, particularly with gold and silver being identified as prominent options for production of nanoparticles. The increasing interest in silver nanoparticles synthesis is due to its multidimensional applications such as water treatment, biosensor, food packaging and preservation, anti-microbial, anti-cancer and anti-inflammatory. Based on its diverse applications, the present study aimed to synthesize silver nanoparticles (SAgNPs) using Spirulina platensis extract as a green source and evaluate their antibacterial, antioxidant, and biocompatible properties. Techniques including UV spectroscopy, scanning electron microscopy (SEM) and Fourier transform-infrared spectroscopy (FTIR) were used to characterize the synthesized SAgNPs. The antioxidant activity was determined through DPPH radical scavenging activity, while the antibacterial activity was assessed using disk diffusion method. Results from UV- spectroscopy revealed a peak absorption at the range of 400–500 nm, with a prominent peak at 423 nm, which confirms the successful biosynthesis of SAgNPs. SEM analysis indicated an average particle size ranging from 50–70 nm. Results from FTIR analysis identified different functional groups (OH-, carbonyl and amino acids) which stabilized the nanoparticles synthesis. Hemolysis assay demonstrated less than 5% hemolysis at all the concentrations studied (100–12.5 µM) that represents the biocompatible nature of SAgNPs. The synthesized SAgNPs showed potent antibacterial activity against both S. aureus and E. coli. Furthermore, the nanoparticles displayed notable antioxidant activity, as evidenced by the antioxidant assay. In conclusion, the biosynthesized SAgNPs derived from Spirulina platensis demonstrated potential antibacterial, antioxidant, and biocompatible properties. These findings highlight the potential of utilizing Spirulina platensis for the biosynthesis of AgNPs with diverse applications in biomedical and pharmaceutical fields. [ABSTRACT FROM AUTHOR]

The free-floating, filamentous cyanobacteria Spirulina platensis is highly valued and in great demand worldwide for its high-value colors and phytonutrients. These compounds find use in health foods, feed, medicines, and diagnostics. The primary objective of this work was to enhance the growth of Spirulina under both oxidative and physiological stress conditions in order to maximize the synthesis of phycocyanin and carotenoid pigments. Cultures were subjected to different concentrations of hydrogen peroxide (H2O2) and sodium chloride for varying temperatures and pH levels. Under lower concentrations of H2O2 (4mM), the maximum carotenoid content was increased, but the phycocyanin content was found to be stimulated at 10mM. Moreover, the synthesis of both pigments was shown to be highest under physiological stress circumstances (40mM and 60mM NaCl concentration) and at an optimal pH of 8-9, helped by a temperature range of 25-30°C. The experimental results demonstrate that both carotenoid and phycocyanin exhibit antioxidant properties even in the presence of oxidative stress. This suggests that the aforementioned circumstances can be applied in future investigations to extract different antioxidants from S. platensis. [ABSTRACT FROM AUTHOR]

Background. Some studies have shown that Spirulina has biological properties exerting beneficial effects on human health. This study aimed to evaluate the effects of Spirulina supplementation on serum glucose, lipid parameters, and oxidative stress markers in male rats with streptozotocin/nicotinamide–induced T2DM. Methods. In this experimental study, rats with diabetes were divided into six groups: healthy control, diabetic control, diabetic-metformin, diabetic-Spirulina 100 mg/dL, diabetic-Spirulina 200 mg/dL, and healthy control-Spirulina 200 mg/d. At the end of the study (28 days), the blood samples were collected, and the fasting blood glucose (FBG), triglycerides (TG), total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), malondialdehyde (MDA), and total antioxidant capacity (TAC) were determined. Results. Significant reductions (P

Abstract Background The Mexican lime (Citrus aurantifolia cv.), widely consumed in Iran and globally, is known for its high perishability. Edible coatings have emerged as a popular method to extend the shelf life of fruits, with xanthan gum-based coatings being particularly favored for their environmental benefits. This study aims to evaluate the effectiveness of an edible coating formulated from xanthan gum, enriched with Spirulina platensis (Sp) and pomegranate seed oil (PSO), in improving the quality and reducing the weight loss of Mexican lime fruit under conditions of 20 ± 2 °C and 50–60% relative humidity. Results Based on the results, the application of coatings was generally effective in reducing fruit weight loss, with the least weight loss observed in the xanthan gum 0.2%+ Spirulina platensis extract (1%) treatment. Additionally, the levels of total phenols and flavonoids in the treated fruits exceeded those in the control group, with xanthan gum 0.2%+ Spirulina platensis extract (1%) and xanthan gum 0.2% exhibiting the highest concentrations of these compounds. The antioxidant capacity of the fruits was also enhanced by the coatings, surpassing that of the control group, with xanthan gum 0.2%+ Spirulina platensis extract (1%) achieving the highest levels. The treatments significantly suppressed the activity of the polyphenol oxidase (PPO) enzyme, with xanthan gum 0.2% demonstrating the most potent inhibitory effect. Furthermore, the treatments resulted in increased activities of catalase (CAT) and peroxidase (POD) enzymes compared to the control. Except for xanthan gum 0.2%+ pomegranate seed oil (0.05%), all treatments maintained the fruit’s greenness (a*) more effectively than the control. Conclusions Peel browning is a major factor contributing to the decline in quality and shelf life of lime fruit. The application of 0.1% and 0.2% xanthan gum coatings, as well as a combination of 0.2% xanthan gum and Spirulina platensis extract, significantly inhibited PPO activity and enhanced the activity of CAT and POD and phenolic compound in Mexican lime fruits stored at of 20 ± 2 °C for 24 days. Consequently, these treatments comprehensively preserved lime fruit quality by significantly reducing browning, maintaining green color, and preserving internal quality parameters such as TA, thereby enhancing both visual appeal and overall fruit quality.

This study examined the protective effects of Spirulina platensis (SP) on African catfish (Clarias gariepinus) against ultraviolet radiation-A (UV-A) induced oxidative stress, immunotoxicity, and histological damage. Recognized for its nutritional value and environmental benefits, SP was evaluated as a potential bioremediation agent. The experiment involved four groups: a control group, a UV-A-exposed group, a UV-A + 100 mg/L SP group, and a UV-A + 200 mg/L SP group, with UV-A exposure for 1 h daily over 3 days. Serum markers of oxidative stress (superoxide dismutase (SOD), total antioxidant capacity (TAC), and malondialdehyde (MDA)) and immune responses (lysozyme (LYZ) and phagocytic activity (PhA)) were measured. UV-A exposure significantly decreased SOD and TAC levels and increased MDA levels. However, SP treatment countered these effects, raising SOD and TAC levels while lowering MDA levels in both SP-treated groups. Similarly, the UV-A-induced reduction in LYZ and PhA activities was reversed by SP treatment, returning to near control levels. Histological analysis showed substantial tissue damage in UV-A-exposed fish, which was mitigated in SP-treated groups, with higher SP concentrations offering greater protection. These results suggest that SP effectively reduces oxidative stress, boosts immune responses, and preserves tissue integrity in UV-A-exposed African catfish. Overall, this study highlights the potential of SP as a valuable bioremediation agent in aquaculture, promoting fish health and resilience to environmental stressors. SP emerges as a promising candidate for enhancing sustainable aquaculture practices through its protective and ameliorative properties. [ABSTRACT FROM AUTHOR]

Abstract The gut microbiota is one of the essential contributors of the pathogenesis and progress of inflammatory bowel disease (IBD). Compared with first-line drug therapy, probiotic supplementation has emerged as a viable and secure therapeutic approach for managing IBD through the regulation of both the immune system and gut microbiota. Nevertheless, the efficacy of oral probiotic supplements is hindered by their susceptibility to the gastrointestinal barrier, leading to diminished bioavailability and restricted intestinal colonization. Here, we developed a bacteria-microalgae symbiosis system (EcN-SP) for targeted intestinal delivery of probiotics and highly effective treatment of colitis. The utilization of mircroalge Spirulina platensis (SP) as a natural carrier for the probiotic Escherichia coli Nissle 1917 (EcN) demonstrated potential benefits in promoting EcN proliferation, facilitating effective intestinal delivery and colonization. The alterations in the binding affinity of EcN-SP within the gastrointestinal environment, coupled with the distinctive structural properties of the SP carrier, served to overcome gastrointestinal barriers, minimizing transgastric EcN loss and enabling sustained intestinal retention and colonization. The oral administration of EcN-SP could effectively treat IBD by reducing the expression of intestinal inflammatory factors, maintaining the intestinal barrier and regulating the balance of gut microbiota. This probiotic delivery approach is inspired by symbiotic interactions found in nature and offers advantages in terms of feasibility, safety, and efficacy, thus holding significant promise for the management of gastrointestinal disorders.

Spirulina (Arthrospira platensis) is a protein-rich source because it contains approximately 60% protein. Therefore, it has positive impacts on human health, including anti-diabetes. The potential of Spirulina platensis as an anti-diabetic agent has been investigated in-vitro and in-vivo. Its extract is capable of inhibiting amylase, α-glucosidase, and DPP IV enzymes in a range of medium activities. Furthermore, spirulina at 250 mg/Kg body weight (BW) in mice can reduce blood glucose levels with similar outcomes to the positive controls (pioglitazone and glibenclamide). Protein groups (Phycocyanin), isolated peptides, and flavonoid groups such as Pinocembrin, Acacetin, and others have been predicted to be responsible for lowering glucose levels. In the market, several commercial products have been sold. Many commercial products have been sold in the market. However, these products are food supplements because a clinical study has not yet been undertaken to confirm their potency and safety. According to this analysis, there is a significant chance to produce anti-diabetic medications from spirulina sources in the future. [ABSTRACT FROM AUTHOR]

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Heat stress (HS) is one of the most significant environmental factors that result in fluctuations and shrinkage in rabbit growth, health, and overall productivity. This study aims to investigate the effects of dietary mineral nanoparticles (selenium or zinc) and/or Spirulina platensis (SP) independently and in combination on stressed growing rabbits. A total of 180 weaned growing New Zealand White rabbits were included in this study and randomly divided into six dietary treatments. Rabbits received a basal diet (control group; CON group) or fortified with SP (1 g/kg diet), selenium nanoparticles (SeNPs, 50 mg/kg diet), zinc nanoparticles (ZnNPs, 100 mg/kg diet), and a mixture of SP and SeNPs (SPSeNPs) or SP and ZnNPs (SPZnNPs) groups for 8 weeks during summer conditions. The obtained results demonstrated a significant increase in the final body weight and weight gain (p < 0.05). Additionally, the feed conversion ratio was improved during the periods from 6 to 14 weeks in the treated rabbits compared to those in the CON group. Dietary supplements considerably improved (p < 0.05) the blood hematology (WBCs, Hb, RBCs, and Hct) and some carcass traits (liver weights and edible giblets). All dietary supplements significantly decreased serum levels of total glycerides (p < 0.0001), AST (p = 0.0113), ALT (p = 0.0013), creatinine (p = 0.0009), and uric acid (p = 0.0035) compared to the CON group. All treated groups (except ZnNPs) had lower values of total bilirubin and indirect bilirubin in a dose-dependent way when compared to the CON group. The values of IgA, IgG, and superoxide dismutase were significantly improved (p < 0.05) in all treated rabbits compared to the CON group. Compared with the CON group, the levels of T3 (p < 0.05) were significantly increased in all treated growing rabbits (except for the ZnNP group), while the serum cortisol, interferon-gamma (IFN-γ), malondialdehyde, and protein carbonyl were significantly decreased in the treated groups (p < 0.05). Dietary supplements sustained the changes in hepatic, renal, and cardiac impairments induced by HS in growing rabbits. Adding SP (1 g/kg diet) or SeNPs (50 mg/kg diet) in the diet, either individually or in combination, improved growth performance, blood picture, and immunity-antioxidant responses in stressed rabbits. Overall, these feed additives (SP, SeNPs, or their mixture) can be applied as an effective nutritional tool to reduce negative impacts of summer stress conditions, thereby maintaining the health status and improving the heat tolerance in growing rabbits. [ABSTRACT FROM AUTHOR]

Monosodium glutamate (MSG), a sodium salt derived from glutamic acid, is widely used in commercial food products to improve taste, quality, and preservation. However, its consumption may have detrimental effects on male reproductive function. Nevertheless, plant extracts, such as Lepidium meyenii (Maca), Trigonella foenum-graecum (Fenugreek), Spirulina platensis (Spirulina), and Tribulus arabica (Tribulus), may ameliorate these adverse effects. To this effect, the phytochemical properties of Lepidium meyenii, Trigonella foenum-graecum, Spirulina platensis, and Tribulus arabica were assessed, and their potential impact on MSG-induced impairment of reproductive parameters was examined. The phytochemical composition (steroids, terpenes, phenols, flavonoids) of the plants was profiled through spectrophotometry and the antioxidant activity was assessed using the 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging assay. Thirty-six male Wistar rats were divided into six groups at random: a control group receiving distilled water, and five experimental groups (MSG, Maca, Fenugreek, Spirulina, and Tribulus) receiving 900 mg/kg/day of MSG dissolved in water for 45 days. Subsequently, the animals in the experimental groups were administered 500 mg/kg/day of the respective plant extract via oral gavage for an additional 35 days, while the MSG group continued to receive water only. Following the treatment period, the animals were sacrificed, and their reproductive tract organs were collected, weighed, and subjected to further analysis. Phytochemical analysis revealed the presence of diverse bioactive elements in the plant extracts, including phenolic and flavonoid compounds. Exposure to MSG negatively impacted total and progressive sperm motility, which was ameliorated by Lepidium meyenii treatment. Sperm morphology showed no significant differences among groups. Treatment of the phytochemical agents diminished histomorphometric alternations of the testicular length, germinal epithelium height, and number of cells in seminiferous tubules, which were caused by the initial administration of MSG. Testosterone and LH levels were reduced in the MSG group but improved in extract-treated groups. The study suggests Lepidium meyenii as a potential remedy for reproductive dysfunction. However, further investigation into its mechanisms and human safety and efficacy is warranted. [ABSTRACT FROM AUTHOR]

Edwardsiellosis is a bacterial fish disease that mostly occurs in freshwater farms and is characterized by a high mortality rate. Edwardsiella tarda strain was recovered from 17 fish out of 50 Nile tilapia, which were harboring clinical signs of systemic septicemia. The level of un-ionized ammonia (NH3) in the fish farm's water was 0.11–0.15 mg/L, which was stressful for the Nile tilapia. Sequencing of the gyrB1 gene confirmed that the isolate was E. tarda JALO4, and it was submitted to NCBI under the accession number PP449014. The isolated E. tarda harbored the virulence gene edw1 AHL-synthase (quorum sensing). In addition, the isolate was sensitive to trimethoprim and sulfamethoxazole mean while it was intermediate to florfenicol. The median lethal dose (LD50) of E. tarda JALO4 was determined to be 1.7 × 105 CFU/mL in Nile tilapia. In the indoor experiment, Nile tilapia (45.05 ± 0.4 g), which received dietary Spirulina platensis (5 and 10 g/kg fish feed), showed optimum growth and feed utilization. Meanwhile, after receiving dietary S. platensis, the fish's feed conversion ratio (FCR) was significantly enhanced compared to the control, which was 1.94, 1.99, and 2.88, respectively. The expression of immune-related genes interleukin (IL)-1β and tumor necrosis factor (TNF)-α were upsurged in E. tarda-challenged fish with higher intensity in S. platensis groups. Dietary S. platensis at a dose of 10 g/kg fish feed could provide a relative protection level (RPL) of 22.2% Nile tilapia challenged against E. tarda. Nile tilapia experimentally infected E. tarda, drastically altering their behavior: higher operculum movement, low food apprehension, and abnormal swimming dietary S. platensis (10 g/kg fish feed) could rapidly restore normal status. It was concluded that Edwardsiellosis could alter Nile tilapia behavior with a high loss in fish population. Fish received dietary-S. platensis could rapidly restore normal behavior after E. tarda infection. It is recommended the incorporation of S. platensis at doses of 10 g/kg into the Nile tilapia diet to boost their immunity and counteract E. tarda infection. [ABSTRACT FROM AUTHOR]

Background: Alzheimer's Disease (AD) is the most prevalent cause of dementia globally and its frequency increases as the world's population ages. AD is one of the largest healthcare issues of the 21st century as the primary cause of dementia and neurodegenerative illnesses. An acquired loss of cognitive function across many cognitive areas is referred to as dementia. The animal studies conducted in this research article to determine whether chemical compounds cause AD may be useful in understanding the disease's mechanism and treatment options. Objectives: The current study assessed the effectiveness of C-Phycocyanin (C-PC) from Spirulina platensis and ethanolic extract of Clerodendreme inerme against scopolamine-induced Alzheimer's in mice. Since Clerodendreme inerme and C-PC are rich in active chemical constituents, they function as beneficial antioxidants. These constituents include saponins, oil, fat, phenolic compounds, tannins, alkaloids, flavonoids, glycosides, terpenoids, steroids, amino acids, proteins and carbs. Thus, an assessment of the anti-Alzheimer properties of the ethanolic extract from Clerodendreme inerme and C-Phycocyanin from Spirulina platensis was conducted. Materials and Methods: The study was designed to evaluate prophylactic effect of Spirulina platensis and Clerodendreme inerme against Scopolamine induced Alzheimer's in mice. Piracetam (200 mg/kg) was used as a standard. The extracts from the plant and leaves of Spirulina platensis and Clerodendreme inerme were screened for anti-alzheimer potential in scopolamine induced Alzheimer's in mice by administering 0.4 mg/kg b.w/day i.p. as test dose. Behavioral Assessments as well as serum levels of AchE, in vitro and Bio-chemical parameters were assessed. Results: Treated with ethanolic extracts of Clerodendreme inerme, C-PC of Spirulina platensis, it demonstrated a dose-dependent reduction in escape latency time and an increase in time spent in the target quadrant. Clerodendreme inerme and C-PC of Spirulina platensis both reduce the escape latency time and transfer latency time in EPM in a dose-dependent manner. AD rodents treated with ethanolic extracts of Clerodendreme inerme, C-PC of Spirulina platensis demonstrated a dose-dependent rise in percentage change in the Y maze test. Apart from behavioral assessments, extracts from ethanolic extract of Clerodendreme inerme, SP demonstrated a dose-dependent decrease in AchE level, since the loss of ACh resulting from AChE's hydrolytic action causes cognitive impairment. AD rodents that were pretreated with ethanolic extract of Clerodendreme inerme, SP there is an increase in catalase and lipid peroxidation. Conclusion: The study concludes that Spirulina platensis and Clerodendreme inerme extracts have a significant anti-Alzheimer activity. This is likely because the extracts contain a variety of nutrients, including saponins, oil, fat, phenolic compounds, tannins, alkaloids, flavonoids, glycosides, terpenoids, amino acids, steroids, proteins and carbs. [ABSTRACT FROM AUTHOR]

Heavy metals such as mercury (Hg) can cause disturbances in the metabolic and phycoremediation activities of microalgae. γ-aminobutyric acid or GABA is a non-protein amino acid and has a signaling role that can regulate the homeostasis of reactive oxygen species (ROS) under environmental stress. In this research, the effect of exogenous GABA on the physiological responses of blue-green algae (cyanobacterium) Spirulina platensis under mercury stress was investigated. HgCl2 in 4 and 8 μM, and GABA in 2 and 4 mM, individually and as a combined treatment, were applied to the microalgal cell suspensions for 48 hours. Then, the indices; dry weight, chlorophyll a, carotenoid, phycobilins, phenol, carbohydrate, protein, malondialdehyde, and polyphenol oxidase and peroxidase activities were evaluated. HgCl2 treatment (individually) decreased dry weight, chlorophyll a, carotenoid, phycoerythrin, phenol, carbohydrate and protein compared to those in control and increased the level of lipid peroxidation and the activities of polyphenol oxidase and peroxidase enzymes. Single treatment of GABA increased dry weight, phycobilin, phenol, malondialdehyde, protein and enzyme activity. But the combined treatments (GABA + HgCl2) improved most of the physiological indices. HgCl2 treatment in one or both concentrations caused a negative effect on the physiological indicators of Spirulina platensis and decreased metabolites such as phenol and proteins, but GABA treatment in most cases reduced the negative effects of mercury and improved the indices. According to the literature, Hg toxicity can be caused by disrupting the biosynthesis of photosynthetic pigments and the activity of enzymes and the occurrence of oxidative stress. While, exogenous GABA can improve the metabolic and physiological cell condition as a signaling molecule through the occurrence of some intracellular events. [ABSTRACT FROM AUTHOR]

Spirulina platensis with bioactive compounds such as Phycocyanin, β-carotene, Genistein and α-glucan, have been shown to have anti-inflammatory properties. Our present study investigation utilizes an in silico methodology to examine the molecular interactions between Spirulina platensis chemicals compound with TNFa and p21. Utilizing computational techniques molecular docking, our goal is to clarify the possible pathways by which Spirulina platensis chemicals could affect these important regulators and postpone cellular senescence. Our research may shed important light on the creation of Spirulina platensis-based therapies for ageing and age-related illnesses. Our results imply that Spirulina platensis may contribute to overall cellular health and the mitigation of cellular senescence. Phycocyanin has the most negative ΔG value is -15.0 kcal/mol. Genistein has the lowest Ki value, namely 7.299 μM. The ΔG and Ki values of Genistein were lower than Quercetin. The potential chemical interactions between substances generated from Spirulina platensis and senescence pathways, including those involving TNFa and p21, are highly intriguing for the development of innovative therapeutic approaches targeted at ameliorating cellular senescence dysfunction associated with aging. [ABSTRACT FROM AUTHOR]

Membrane photobioreactors (MPBRs) have gained significant attention due to their ability to support microalgae activities such as cultivation, harvesting, and production of beneficial products. Despite various efforts to mitigate membrane fouling, a fundamental issue in membrane processes, in these systems, a cost-effective and less energy-consuming method is still needed. This study examines the impact of the cross-sectional area of the riser and the baffle material on membrane fouling in an internal loop airlift MPBR. The use of hydrophilic polyester-polypropylene (PES–PP) baffles proves to be more effective than plexiglass baffles. Specifically, in configurations with d = 0.7 cm and d = 1.4 cm, RC/RT decreased by approximately 20% and 13%, respectively, compared to plexiglass baffles. As for the values of RP/RT at a distance of d = 0.7, nearly a 5% increase was observed, and at a distance of d = 1.4, an increase of approximately 11% was observed. This is due to the development of the cake layer on the matrix structure of the PES–PP baffles instead of the membrane itself. The most optimal outcomes were reached while working with PES–PP at a distance of 0.7 cm, as it prolonged the membrane fouling time to 46 h. [ABSTRACT FROM AUTHOR]

Background: The production of secondary metabolites in different strains of microalgae varies and is likely dependent on environmental conditions. Consequently, the production of bioactive substances as secondary metabolites occurs in microalgae to aid their survival in adverse environmental conditions such as salinity stress. Objectives: The purpose of this study is to investigate the antibacterial activity of the methanolic extract of Spirulina platensis algae cultivated under different salinity stresses against Yersinia rukeri, Escherichia coli, Salmonella sp., and Vibrio cholerae. Methods: The broth microdilution method was used to evaluate the growth inhibitory activity of the extracts against Yersinia ruckeri, Salmonella sp., Escherichia coli, and Vibrio cholerae bacteria. The minimum inhibitory concentration (MIC) and the minimum bactericidal concentration (MBC) were determined by this method, and the diameter of the inhibition zone was assessed through the well diffusion method. Results: In the treatment with algae extract grown under 3.5 ppt salinity stress, the concentrations of 50 and 25 mg/mL extract showed the largest inhibition zone diameter against Salmonella sp. compared to other bacteria. Under 7 ppt salinity stress, all studied concentrations of algal extract exhibited a higher inhibition zone diameter against V. cholerae compared to other bacteria. The results of comparing different concentrations of algae extract between the two salinity stresses of 3.5 and 7 ppt for each bacterium showed a significant difference (P < 0.05). As salinity increased, the diameter of the inhibition zone also increased for all bacteria. Conclusions: Our results showed an increase in the antibacterial activity of the methanolic extract of Spirulina platensis with higher salinity stress levels. Therefore, cultivating Spirulina algae in salt water can be a cost-effective and suitable method to produce more secondary metabolites for use in the pharmaceutical industry. [ABSTRACT FROM AUTHOR]

This study was conducted at El-Sabahia Research Station (latitude 31°12′ N, longitude 29°58′ E) in Alexandria, Egypt to evaluate the effect of Spirulina platensis algae extract on the growth, yield, and juice quality of four sugarcane genotypes during the 2020/2021 and 2021/2022 seasons. The sugarcane genotypes tested (G.T. 54-9, G. 2003-47, G. 84-47, and G. 2004-27) were treated with four concentrations (0, 0.1, 0.2, and 0.3%) of spirulina algae extract (SE) during their development as plant cane and first ratoon crops. At harvest, the growth, physiological, and juice quality characteristics were documented, while relative chlorophyll content was measured 210 days after sowing. Spraying canes with 0.2% of SE was the most effective treatment in enhancing all of the evaluated characteristics compared to those left without SE. Cane yield was more closely correlated with stalk weight (r = 0.88), followed by leaf area index (r = 0.82), relative chlorophyll content (r = 0.82), stalk length (r = 0.76), and number of tillers (r = 0.73), while recoverable sugar yield was closely correlated (p < 0.01) with sugar content % (r = 0.76). [ABSTRACT FROM AUTHOR]

Microalgae can enhance the nutritional value of conventional foods and hence to positively affect human health, due to their original chemical composition. The present study aimed to produce functional Karish cheese fortified with different types and ratios of microalgae powder (Chlorella vulgaris and Spirulina platensis) and evaluate the role of fortification with those algae on the quality of the product. Karish cheese was incorporated with C. vulgaris and S. platensis powder, separately at three levels 0.5, 1.0 and 1.5%. The evaluation of chemical composition, salt content, color parameters, antioxidant activity, total phenolic and total carotenoids content, texture profile and organoleptic properties of Karish cheese fortified with microalgae were determined. Addition of microalgae to Karish cheese showed an increasing (P<0.05) in total solid content, protein, ash, acidity and enhanced the texture profile in the final product compared with control sample. Also, the results indicated highest content of antioxidant activity, total flavonoid and total phenolic among all treatments compared with control. Organoleptic preferred Karish cheese samples enriched with microalgae especially with 0.5 and 1.0 % level consequently. Based on these results microalgae powder enriched cheese may have a great benefit for the industry to improve the texture, shapes and the functional nutraceutical properties of the products. [ABSTRACT FROM AUTHOR]

In the present study, fortification of industrial noodles with lentil flour (7-35 %), soy protein isolate (SPI), albumin and Spirulina platensis microalgae were (at 1 to 5 %) and their physicochemical, rheological and sensory properties were evaluated. Treatments containing 1-5 % of S.platensis had higher ash, moisture, fat and protein content compared to treatments containing albumin, lentil flour, soy isolate content and control samples. Increasing the levels ofS.platensis and lentil flour individually resulted in a decrease in the peroxide value. The lowest L*, a* and b* value were related to the treatments containing S.platensis, while the treatments containing lentil flour had the highest a* and b* and the highest a* was related to the treatments containing albumin. In all treatments, with increasing the level of all additives, the firmness increased and treatments containing SPI showed the highest firmness. However, the highest adhesion, gumminess and chewability were related to the treatments containing lentil flour and the highest gumminess and chewability were related to albumin. Different levels of albumin had the highest color scores compared to other treatments and control samples. The highest score of taste, texture and overall acceptance belonged to the treatments containing different levels (2 - 5%) of albumin. As a conclusion, it can be said that different levels of albumin had the most significant effect on the quality properties of industrial noodle samples, and in contrast, treatments containing spirulina did not show acceptable results in the quality tests evaluated. [ABSTRACT FROM AUTHOR]

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The booming aquaculture industry has created a strong demand for fishmeal and increased environmental pressures. Spirulina, as a potential alternative to fishmeal, has been shown to have growth-promoting and animal health-enhancing properties. In this study, 600 large spiny loaches, divided into five experimental groups, F0, F1, F2, F3, and F4, were reared for 10 weeks using Spirulina platensis powder (SPP) as a substitute for 0%, 5%, 10%, 15%, and 20% of fishmeal, respectively. The results of intestinal physiological indexes showed that superoxide dismutase was lower than F0 in all treatment groups, and the activity of F3 was significantly lower than F0 (p < 0.05). The activity of malondialdehyde was significantly higher than that of F0 in all groups except F3 (p < 0.05). The addition of SPP also led to a decrease in the activity of acid phosphatase in the intestine, which was significantly lower in all treatment groups compared to the F0 group (p < 0.05). The results of serum physiology showed that the activity of superoxide dismutase in serum gradually increased with the increase in the percentage of SPP addition, and the F3 group produced a significant difference from the F0 group (p < 0.05). The transcriptomics results showed that DEGs in the low percentage substitution group (<15%) were mostly enriched in metabolism-related pathways, such as bile secretion; DEGs in the high percentage substitution group (>15%) were mostly enriched in inflammation-related pathways, such as complement p and coagulation cascades. Metabolomics confirmed that nicotinate and nicotinamide metabolism and glycerophospholipid metabolism were the two pathways that were significantly enriched in the treatment groups of fishmeal replacement by SPP. The present study demonstrated that a low percentage (<15%) of fishmeal replacement by SPP in feed mobilized MA digestive metabolism, whereas a high percentage (>15%) of replacement induced intestinal stress. Considering the health and farm efficiency aspects, the proportion of SPP in feed formulation for MA should be less than 15%. [ABSTRACT FROM AUTHOR]

The whole-cell inorganic-biohybrid systems show special functions and wide potential in biomedical application owing to the exceptional interactions between microbes and inorganic materials. However, the hybrid systems are still in stage of proof of concept. Here, we report a whole-cell inorganic-biohybrid system composed of Spirulina platensis and gold nanoclusters (SP-Au), which can enhance the cancer radiotherapy through multiple pathways, including cascade photocatalysis. Such systems can first produce oxygen under light irradiation, then convert some of the oxygen to superoxide anion (•O2−), and further oxidize the glutathione (GSH) in tumor cells. With the combination of hypoxic regulation, •O2− production, GSH oxidation, and the radiotherapy sensitization of gold nanoclusters, the final radiation is effectively enhanced, which show the best antitumor efficacy than other groups in both 4T1 and A549 tumor models. Moreover, in vivo distribution experiments show that the SP-Au can accumulate in the tumor and be rapidly metabolized through biodegradation, further indicating its application potential as a new multiway enhanced radiotherapy sensitizer. [ABSTRACT FROM AUTHOR]

The gut microbiota is one of the essential contributors of the pathogenesis and progress of inflammatory bowel disease (IBD). Compared with first-line drug therapy, probiotic supplementation has emerged as a viable and secure therapeutic approach for managing IBD through the regulation of both the immune system and gut microbiota. Nevertheless, the efficacy of oral probiotic supplements is hindered by their susceptibility to the gastrointestinal barrier, leading to diminished bioavailability and restricted intestinal colonization. Here, we developed a bacteria-microalgae symbiosis system (EcN-SP) for targeted intestinal delivery of probiotics and highly effective treatment of colitis. The utilization of mircroalge Spirulina platensis (SP) as a natural carrier for the probiotic Escherichia coli Nissle 1917 (EcN) demonstrated potential benefits in promoting EcN proliferation, facilitating effective intestinal delivery and colonization. The alterations in the binding affinity of EcN-SP within the gastrointestinal environment, coupled with the distinctive structural properties of the SP carrier, served to overcome gastrointestinal barriers, minimizing transgastric EcN loss and enabling sustained intestinal retention and colonization. The oral administration of EcN-SP could effectively treat IBD by reducing the expression of intestinal inflammatory factors, maintaining the intestinal barrier and regulating the balance of gut microbiota. This probiotic delivery approach is inspired by symbiotic interactions found in nature and offers advantages in terms of feasibility, safety, and efficacy, thus holding significant promise for the management of gastrointestinal disorders. [ABSTRACT FROM AUTHOR]

The study was conducted to study whether feeding Spirulina platensis has beneficial effect on enhancing the fatty acid profile and storage stability of the loin muscle of pigs. A total of 24 male finishing pigs (Large White Yorkshire X Desi) were housed individually and distributed randomly into four equal groups with six animals in each group. Four isonitrogenous experimental diets (T1 to T4) were formulated by including dry Spirulina platensis at 0, 0.5, 1.0 and 1.5 %, respectively and fed during finishing phase (50 to 75 kg) live weight. Dietary treatments had no significant (P>0.05) effect on the total lipid and total cholesterol content of the loin muscle of pigs. The level of polyunsaturated fatty acids (PUFA) was higher (P<0.05) in Spirulina supplemented groups (T2, T3, and T4) compared to T1 leading to a higher (P<0.05) ratio of PUFA to saturated fatty acid (SFA) with increasing levels of Spirulina platensis inclusion in the diet. Whereas, the sum of SFA and monounsaturated fatty acids (MUFAs) was significantly (P<0.05) decreased in Spirulina supplemented groups compared to T1. The Ratio of n-6 to n-3 of muscle was decreased significantly (P<0.05) with increasing level of Spirulina and the values were 21.5, 20.2, 20.1, and 19.8 in T1 to T4, respectively. The other fatty acids namely gamma-linolenic acid, alpha-linolenic acid and eicosapentanoic acid were not statistically significant (P>0.05) among the treatment groups. Supplementation of Spirulina significantly (P<0.05) decreased the pH and 2-TBARS value of loin muscle during the refrigerated (4±1°C) storage at 0, 3, 6, and 9 days. Further, inclusion of Spirulina showed significant (P<0.05) improvement in the sensory attributes (flavour, tenderness, and juiciness) of the loin muscle of pigs. The results indicated that inclusion of dried Spirulina platensis powder up to 1.5 % in finishing pig diets was beneficial to achieve PUFA enriched pork and overall acceptability of the pork during refrigerated storage for a period of 9 days. [ABSTRACT FROM AUTHOR]

Spirulina platensis microalga has become recognized as a promising source of highly nutritious food components to feed the growing global population. Because of its high protein content, abundance of essential amino acids, and excellent digestion, it is employed in human nutrition and there is growing interest in analyzing bioactive compound present in Spirulina platensis microalga. In this work, a portable near-infrared (NIR) spectrometer was used for the monitoring of physicochemical properties (extraction yield (EY), total polyphenols concentration (TPC), total proteins concentration (TP), antioxidant activity measured by (i) the DPPH method (DPPH) and (ii) FRAP method (FRAP)) of Spirulina platensis aqueous extracts. The ultrasound-assisted aqueous extraction (ultrasonic bath with an ultrasound frequency of 35 kHz) of bioactive molecules from Spirulina platensis was performed throughout 30 independent experiments. NIR spectra were recorded in the wavelength range of 900–1700 nm. Raw NIR spectra were subjected to the physicochemical properties applying Principal Component Analysis (PCA), partial least square (PLS), and artificial neural network (ANN) modeling. Results show that ANN models developed for the prediction of TPC and DPPH can be utilized for extraction process control (RER > 10), while the other three models can be employed for screening (RER > 4). Generally, the obtained results indicate significant potential for using portable NIR spectroscopy for the analysis of Spirulina platensis aqueous extracts. [ABSTRACT FROM AUTHOR]

Spirulina platensis (SP) has gained popularity over the last few years, owing to its remarkable nutritional properties and high potential across various industrial sectors. In this study, we analyzed the volatile profile of eight SP samples from the same strain subjected to different drying (oven-drying, air-drying, and spray-drying) and storing conditions ("freshly prepared" and after 12 months of storage) using HS-SPME-GC-MS. Principal component analysis (PCA) was used as a multivariate technique to discern similarities and differences among the samples. The main aim was to assess the impact of the drying technique on the aroma profile and storage life of SP samples. Air-drying leads to the less pronounced formation of by-products related to heat treatment, such as Maillard and Strecker degradation compounds, but promotes oxidative and fermentative phenomena, with the formation of organic acids and esters, especially during storage. Thermal treatment, essential for limiting degradation and fermentation during storage and extending shelf life, alters the aroma profile through the formation of volatile compounds, such as Strecker aldehydes and linear aldehydes, from amino acid and lipid degradation. High temperatures in spray-drying favor the formation of pyrazines. The findings underscore the trade-offs inherent in choosing an appropriate drying method, thereby informing decision-making processes in industrial settings aimed at optimizing both product quality and efficiency. [ABSTRACT FROM AUTHOR]