RESUMO
The intricate composition of microalgal pigments plays a crucial role in various biological processes, from photosynthesis to biomarker identification. Traditional pigment analysis methods involve complex extraction techniques, posing challenges in maintaining analyte integrity. In this study, we employ Electron Transfer Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry (ET-MALDI-MS) to compare the pigmentary profiles of Chlorella vulgaris intact cells, chloroplasts, and solvent extracts. We aim to obtain comprehensive extracts rich in polar and non-polar compounds using ultrasound-assisted and supercritical fluid extraction methods. Additionally, intact chloroplasts are isolated using a lysis buffer and sucrose density gradient centrifugation. Our ET-MALDI-MS analysis reveals distinct compositional differences, highlighting the impact of extraction protocols on microalgal pigment identification. We observe prominent signals corresponding to radical cations of key pigments, including chlorophylls and carotenoids, which are crucial for C. vulgaris identification. Furthermore, ET-MALDI-MS facilitates the identification of specific lipids within chloroplast membranes and other organelles. This study underscores the rapid and precise nature of ET-MALDI-MS in microalgal biomarker analysis, providing valuable insights into phytoplankton dynamics, trophic levels, and environmental processes. C. vulgaris emerges as a promising model for studying pigment composition and membrane lipid diversity, enhancing our understanding of microalgal ecosystems.
Assuntos
Chlorella vulgaris , Cloroplastos , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Chlorella vulgaris/química , Chlorella vulgaris/metabolismo , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/métodos , Cloroplastos/química , Cloroplastos/metabolismo , Carotenoides/análise , Carotenoides/química , Clorofila/análise , Clorofila/química , Pigmentos Biológicos/análise , Pigmentos Biológicos/químicaRESUMO
The present study evaluated the hematological, antiparasitic and growth responses in tambaqui (Colossoma macropomum) fed with diets supplemented with the microalgae Arthrospira platensis and Chlorella vulgaris (0%; 10% A. platensis; 10% C. vulgaris; and 5% A. platensis+5% C. vulgaris). Tambaqui (n=60, 62.57 ± 8.76 g) were fed for 20 days with experimental diets. Blood samples collection was done to determine hematological parameters, and gills were removed to identify and count monogenetic parasites. Supplementation with A. platensis 10% reduced red blood cells count, in consequence mean corpuscular volume and mean hemoglobin concentration increased. Total leukocyte, monocyte, eosinophil, and basophil counts reduced with the use of A. platensis. Higher monocytes, eosinophil, and basophil numbers in tambaqui fed with diet supplemented with 10% C. vulgaris were observed and may have been due to the presence of immunostimulants in this microalga composition. Reduction on total cholesterol in tambaqui that received both microalgae (A. platensis 5%+C. vulgaris 5%) may indicate that combined supplementation presented greater benefits to the health for C. macropomum than separately. Both microalgae were efficient against monogenetic parasites of tambaqui. Thus, the dietary use of the microalgae A. platensis and C. vulgaris provided immunostimulant and antiparasitic efficacy in C. macropomum.
Assuntos
Chlorella vulgaris , Spirulina , Chlorella vulgaris/química , Animais , Suplementos Nutricionais , Caraciformes , Microalgas/químicaRESUMO
Background: Microbial Fuel Cell (MFC) technology is used in various applications such as wastewater treatment with the production of electrical energy. The objective of this study was to estimate the biodepuration of oils and fats, the elimination of blue dye brl and bioelectro-characterization in MFCs with Chlorella vulgaris and bacterial community. Results: The operation of MFCs at 32 d showed an increase in bioelectrogenic activity (from 23.17 to 327.67 mW/m2 ) and in the potential (from 200 to 954 mV), with biodepuration of fats and oils (95%) in the microalgal cathode, and a removal of the chemical oxygen demand COD (anode, 71%, cathode, 78.6%) and the blue dye brl (73%) at the anode, here biofilms were formed by the bacterial community consisting of Actinobacteria and Deltaproteobacteria. Conclusions: These findings suggest that MFCs with C. vulgaris and bacterial community have a simultaneous efficiency in the production of bioelectricity and bioremediation processes, becoming an important source of bioenergy in the future.
Assuntos
Bactérias/metabolismo , Fontes de Energia Bioelétrica/microbiologia , Purificação da Água/métodos , Chlorella vulgaris/metabolismo , Bactérias/química , Biofilmes , Chlorella vulgaris/química , Eletricidade , Eletrodos , Microalgas , Eletroforese em Gel de Gradiente Desnaturante , Águas ResiduáriasRESUMO
BACKGROUND: From ancient times, marine algae have emerged as alternative medicine and foods, contains the rich source of natural products like proteins, vitamins, and secondary metabolites, especially Chlorella vulgaris (C. vulgaris) contains numerous anti-inflammatory, antioxidants and wound healing substances. Type 2 diabetes mellitus is closely associated with adipogenesis and their factors. Hence, we aimed to investigate the chemical constituents and adipogenic modulatory properties of C. vulgaris in 3T3-L1 pre-adipocytes. RESULTS: We analysed chemical constituents in ethanolic extract of C. vulgaris (EECV) by LC-MS. Results revealed that the EECV contains few triterpenoids and saponin compounds. Further, the effect of EECV on lipid accumulation along with genes and proteins expressions which are associated with adipogenesis and lipogenesis were evaluated using oil red O staining, qPCR and western blot techniques. The data indicated that that EECV treatment increased differentiation and lipid accumulation in 3T3-L1 cells, which indicates positive regulation of adipogenic and lipogenic activity. These increases were associated with up-regulation of PPAR-γ2, C/EBP-α, adiponectin, FAS, and leptin mRNA and protein expressions. Also, EECV treatments increased the concentration of glycerol releases as compared with control cells. Troglitazone is a PPAR-γ agonist that stimulates the PPAR-γ2, adiponectin, and GLUT-4 expressions. Similarly, EECV treatments significantly upregulated PPAR-γ2, adiponectin, GLUT-4 expressions and glucose utilization. Further, EECV treatment decreased AMPK-α expression as compared with control and metformin treated cells. CONCLUSION: The present research findings confirmed that the EECV effectively modulates the lipid accumulation and differentiation in 3T3-L1 cells through AMPK-α mediated signalling pathway.
Assuntos
Células 3T3-L1/efeitos dos fármacos , Chlorella vulgaris/química , Extratos Vegetais/farmacologia , Alga Marinha/química , Células 3T3-L1/fisiologia , Proteínas Quinases Ativadas por AMP/análise , Proteínas Quinases Ativadas por AMP/efeitos dos fármacos , Proteínas Quinases Ativadas por AMP/metabolismo , Adipócitos/citologia , Adipócitos/efeitos dos fármacos , Adipócitos/metabolismo , Adiponectina/análise , Adiponectina/metabolismo , Animais , Diferenciação Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Diabetes Mellitus Tipo 2/metabolismo , Regulação para Baixo , Expressão Gênica , Glucose/metabolismo , Transportador de Glucose Tipo 4/análise , Transportador de Glucose Tipo 4/efeitos dos fármacos , Transportador de Glucose Tipo 4/metabolismo , Camundongos , PPAR gama/análise , PPAR gama/efeitos dos fármacos , PPAR gama/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Fatores de Tempo , Regulação para CimaRESUMO
In the present investigation, the application of NMR spectroscopic techniques was extensively used with an objective to explore the biodiesel potential of biomass cultivated on a lab scale using strains of Chlorella vulgaris and Scenedesmus ecornis. The effect of variation in the composition of culturing medium on the neutral and polar lipids productivity, and fatty acid profile of solvent extracts of microalgae biomass was studied. Determination of unsaturated fatty acid composition (C18:N = 1-3, ω3 C20:5, ω3 C22:6), polyunsaturated fatty esters (PUFEs), saturated fatty acids (SFAs), unsaturated fatty acids (UFAs), free fatty acids (FFAs), and iodine value were achieved from a single (1)H NMR spectral analysis. The results were validated by (13)C NMR and GC-MS analyses. It was demonstrated that newly developed methods based on (1)H and (13)C NMR techniques are direct, rapid, and convenient for monitoring the microalgae cultivation process for enhancement of lipid productivity and their quality aspects in the solvent extracts of microalgal biomasses without any sample treatment and prior separation compared to other methods. The fatty acid composition of algae extracts was found to be similar to vegetable and fish oils, mostly rich in C16:0, C18:N (N = 0 to 3), and n-3 omega polyunsaturated fatty acids (PUFAs). The lipid content, particularly neutral lipids, as well as most of the quality parameters were found to be medium specific by both the strains. The newly developed methods based on NMR and ultrasonic procedure developed for efficient extraction of neutral lipids are cost economic and can be an effective aid for rapid screening of algae strains for modulation of lipid productivity with desired biodiesel quality and value-added products including fatty acid profile.
Assuntos
Chlorella vulgaris/química , Chlorella vulgaris/metabolismo , Lipídeos/análise , Espectroscopia de Ressonância Magnética/métodos , Scenedesmus/química , Scenedesmus/metabolismo , Biocombustíveis , Biomassa , Chlorella vulgaris/crescimento & desenvolvimento , Meios de Cultura/química , Cromatografia Gasosa-Espectrometria de Massas , Scenedesmus/crescimento & desenvolvimento , Sonicação/métodosRESUMO
BACKGROUND: From ancient times, marine algae have emerged as alternative medicine and foods, contains the rich source of natural products like proteins, vitamins, and secondary metabolites, especially Chlorella vulgaris (C. vulgaris) contains numerous anti-inflammatory, antioxidants and wound healing substances. Type 2 diabetes mellitus is closely associated with adipogenesis and their factors. Hence, we aimed to investigate the chemical constituents and adipo-genic modulatory properties of C. vulgaris in 3T3-L1 pre-adipocytes. RESULTS: We analysed chemical constituents in ethanolic extract of C. vulgaris (EECV) by LC-MS. Results revealed that the EECV contains few triterpenoids and saponin compounds. Further, the effect of EECV on lipid accumulation along with genes and proteins expressions which are associated with adipogenesis and lipogenesis were evaluated using oil red O staining, qPCR and western blot techniques. The data indicated that that EECV treatment increased differentiation and lipid accumulation in 3T3-L1 cells, which indicates positive regulation of adipogenic and lipogenic activity. These increases were associated with up-regulation of PPAR-γ2, C/EBP-α, adiponectin, FAS, and leptin mRNA and protein expressions. Also, EECV treatments increased the concentration of glycerol releases as compared with control cells. Troglitazone is a PPAR-γ agonist that stimulates the PPAR-y2, adiponectin, and GLUT-4 expressions. Similarly, EECV treatments significantly upregulated PPAR-γ, adiponectin, GLUT-4 expressions and glucose utilization. Further, EECV treatment decreased AMPK-α expression as compared with control and metformin treated cells. CONCLUSION: The present research findings confirmed that the EECV effectively modulates the lipid accumulation and differentiation in 3T3-L1 cells through AMPK-α mediated signalling pathway.
Assuntos
Animais , Camundongos , Alga Marinha/química , Extratos Vegetais/farmacologia , Células 3T3-L1/efeitos dos fármacos , Chlorella vulgaris/química , Fatores de Tempo , Regulação para Baixo , Expressão Gênica , Diferenciação Celular/efeitos dos fármacos , Regulação para Cima , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Adipócitos/citologia , Adipócitos/efeitos dos fármacos , Adipócitos/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Células 3T3-L1/fisiologia , PPAR gama/análise , PPAR gama/efeitos dos fármacos , PPAR gama/metabolismo , Diabetes Mellitus Tipo 2/metabolismo , Adiponectina/análise , Adiponectina/metabolismo , Transportador de Glucose Tipo 4/análise , Transportador de Glucose Tipo 4/efeitos dos fármacos , Transportador de Glucose Tipo 4/metabolismo , Proteínas Quinases Ativadas por AMP/análise , Proteínas Quinases Ativadas por AMP/efeitos dos fármacos , Proteínas Quinases Ativadas por AMP/metabolismo , Glucose/metabolismoRESUMO
Objectives: To describe acute toxicity, antibacterial activity and phytochemical assessment of Chlorella vulgaris and Spirulina platensis powders. Material and Methods: FeCl3 test, Wagner test, Keller Killiani test, frothing test, alkaline solution and dilute acid; concentrated sulphuric acid were used for phytochemical analysis. Antibacterial screening of the extracts was conducted using the disc gel diffusion method in E. coli, S. aureus and B. cereus clinical strains. In order to evaluate acute toxicity and its effects on haematological and biochemical parameters; 15 albino rats were grouped into five groups: I (powder of aqueous extract of Chlorella vulgaris), II (powder of methanol extract of Chlorella vulgaris), III (powder of aqueous extract of Spirulina platensis), IV (powder of methanol extract of Spirulina platensis) and V (control). The dosage was 25g/day/rat. After six days, haematological and biochemical parameters and gross pathologic changes were evaluated. Results: Alkaloids and flavonoids were detected from the methanol extracts of both Chlorella vulgaris and Spirulina platensis (Arthrospira). Only cardiac glycosides and steroids were detected from Spirulina's extracts. Chlorella vulgaris extracts significantly inhibited B. cereus. Rats fed with Chlorella vulgaris and Spirulina platensis powder showed an increase in white blood cell counts and haemoglobin level compared to negative control rats (p<0.001). Serum glumatic oxalate transaminase (SGOT) and serum glutamate pyruvate transaminase (SGPT) had normal values but significative differences between groups (p<0.001). Conclusion: This powder is rich in bioactive phytochemicals but only Chlorella's extracts have shown antibacterial effect. Signs of toxicity weren't found in any parameter.
Objetivos: Describir la toxicidad aguda, efecto antibacteriano y análisis fitoquímico de los polvos de Chlorella vulgaris y Spirulina platensis. Materiales y métodos: Se realizaron las pruebas de FeCl3, Keller Killiani, de saponinas, solución alcalina y de concentración de ácido sulfúrico para el análisis fitoquímico. El efecto antibacteriano de los extractos fue evaluado mediante el método de difusión con discos en cepas de E. coli, S. aureus y B. cereus. Para evaluar la toxicidad aguda y los efectos del polvo en parámetros hematológicos y bioquímicos, se agruparon 15 ratas albinas en cinco grupos: I (polvo de extracto acuoso de Chlorella vulgaris); II (polvo de extracto metanólico de Chlorella vulgaris); III (polvo de extracto acuoso de Spirulina platensis); IV (polvo de extracto metanólico de Spirulina platensis), y V (grupo control). La dosis usada fue de 25 g/día/rata. Después de seis días, se evaluaron todos los parámetros y cambios macroscópicos en los órganos. Resultados: Se encontraron alcaloides y flavonoides en los extractos metanólicos de Chlorella vulgaris y Spirulina platensis (Arthrospira). Se detectaron glucósidos cardiacos y esteroides en los extractos de Spirulina platensis. Los extractos de Chlorella vulgaris inhibieron el crecimiento de Bacillus cereus. Las ratas alimentadas con los polvos de Chlorella vulgaris y Spirulina platensis incrementaron el conteo de leucocitos y los valores de hemoglobina comparados con el grupo control (p<0,001). Las transaminasas (SGOT y SGPT) se encontraron en valores normales, pero con diferencias significativas entre los grupos (p<0,001). Conclusiones: Estos polvos son ricos en componentes fitoquímicos activos, pero solo los extractos de Chorella vulgaris mostraron efecto antibacteriano. No se encontraron signos de toxicidad aguda en ningún parámetro.
Assuntos
Animais , Ratos , Chlorella vulgaris/química , Testes de Sensibilidade a Antimicrobianos por Disco-Difusão , Compostos Fitoquímicos , Extratos Vegetais/toxicidade , Modelos AnimaisRESUMO
The gasification of microalgae Chlorella vulgaris under an atmosphere of argon and water vapor was investigated by thermogravimetric analysis. The data were interpreted by using conventional isoconversional methods and also by the independent parallel reaction (IPR) model, in which the degradation is considered to happen individually to each pseudo-component of biomass (lipid, carbohydrate and protein). The IPR model allows obtaining the kinetic parameters of the degradation reaction of each component. Three main stages were observed during the gasification process and the differential thermogravimetric curve was satisfactorily fitted by the IPR model considering three pseudocomponents. The comparison of the activation energy values obtained by the methods and those found in the literature for other microalgae was satisfactory. Quantification of reaction products was performed using online gas chromatography. The major products detected were H2, CO and CH4, indicating the potential for producing fuel gas and syngas from microalgae.
Assuntos
Biocombustíveis , Chlorella vulgaris/química , Microalgas/química , Biomassa , Cromatografia Gasosa , Cinética , Modelos Químicos , TermogravimetriaRESUMO
Three lipid extraction methods of hexane Soxhlet (Sox-Hex), Halim (HIP), and Bligh and Dyer (BD) were applied on freeze-dried (FD) and oven-dried (OD) Chlorella vulgaris biomass to evaluate their effects on lipid yield, fatty acid profile, and algal biodiesel quality. Among these three methods, HIP was the preferred one for C. vulgaris lipid recovery considering both extraction efficiency and solvent toxicity. It had the highest lipid yields of 20.0 and 22.0% on FD and OD biomass, respectively, with corresponding neutral lipid yields of 14.8 and 12.7%. The lipid profiling analysis showed that palmitic, oleic, linoleic, and α-linolenic acids were the major fatty acids in the algal lipids, and there were no significant differences on the amount of these acids between different drying and extraction methods. Correlative models applied to the fatty acid profiles concluded that high contents of palmitic and oleic acids in algal lipids contributed to balancing the ratio of saturated and unsaturated fatty acids and led to a high-quality algal biodiesel.
Assuntos
Biocombustíveis/análise , Fracionamento Químico/métodos , Chlorella vulgaris/metabolismo , Ácidos Graxos/química , Ácidos Graxos/isolamento & purificação , Liofilização/métodos , Biomassa , Chlorella vulgaris/química , Chlorella vulgaris/crescimento & desenvolvimento , Ácidos Graxos/metabolismo , Lipídeos/química , Lipídeos/isolamento & purificaçãoRESUMO
AIMS: The search for natural agents that minimize obesity-associated disorders is receiving special attention. In this regard, the present study aimed to evaluate the prophylactic effect of Chlorella vulgaris (CV) on body weight, lipid profile, blood glucose and insulin signaling in liver, skeletal muscle and adipose tissue of diet-induced obese mice. MAIN METHODS: Balb/C mice were fed either with standard rodent chow diet or high-fat diet (HFD) and received concomitant treatment with CV for 12 consecutive weeks. Triglyceride, free fatty acid, total cholesterol and fractions of cholesterol were measured using commercial assay. Insulin and leptin levels were determined by enzyme-linked immunosorbent assay (ELISA). Insulin and glucose tolerance tests were performed. The expression and phosphorylation of IRß, IRS-1 and Akt were determined by Western blot analyses. KEY FINDINGS: Herein we demonstrate for the first time in the literature that prevention by CV of high-fat diet-induced insulin resistance in obese mice, as shown by increased glucose and insulin tolerance, is in part due to the improvement in the insulin signaling pathway at its main target tissues, by increasing the phosphorylation levels of proteins such as IR, IRS-1 and Akt. In parallel, the lower phosphorylation levels of IRS-1(ser307) were observed in obese mice. We also found that CV administration prevents high-fat diet-induced dyslipidemia by reducing triglyceride, cholesterol and free fatty acid levels. SIGNIFICANCE: We propose that the modulatory effect of CV treatment preventing the deleterious effects induced by high-fat diet is a good indicator for its use as a prophylactic-therapeutic agent against obesity-related complications.