Evaluación del potencial de Desmodesmus asymmetricus y Chlorella vulgaris para la remoción de nitratos y fosfatos de aguas residuales / Potential assessment of Desmodesmus asymmetricus and Chlorella vulgaris for removal of nitrates and phosphates in wastewater

Resumen En este estudio evaluamos las microalgas Chlorella vulgaris y Demodesmus asymmetricus, con el fin de determinar la proporción de microalgas que remueven la mayor concentración de nitrógeno y fósforo en aguas residuales de una planta de tratamiento. Se realizó un diseño de mezclas de las microalgas con el agua residual durante 9 días, con fotoperiodo 12:12 h, en un invernadero. Las mayores eficiencias de remoción de nitrógeno (principalmente NO3 -N y NO2 -N) y fósforo (PO4 ) se observaron en los tratamientos con mayor proporción de D. asymmetricus, entre ellos sobresalió el tratamiento T3 (25% C. vulgaris / 75% D. asymmetricus) donde se removió el 100% de nitrógeno y 77.1% de fósforo. De igual manera con el oxígeno disuelto (OD), el T3 obtuvo el mejor resultado alcanzando una media de 25.50 ± 0.28 mg/L. Finalmente, de acuerdo con el análisis del diseño de mezclas, se determinó que la mezcla óptima de microalgas que logra la mayor producción de OD y la mayor remoción, fue la proporción de 6% de C. vulgaris y 94% de D. asymmetricus en un tiempo de cultivo de 9 días. En conclusión, las microalgas demuestran su capacidad de biorremediación de aguas residuales domésticas.

Abstract In this work, we evaluated the proportion Chlorella vulgaris and Demodesmus asymmetricus microalgae, that removes the highest concentration of nitrogen and phosphorus in wastewater from a treatment plant. A mixture design was employed in this study, for 9 days, with 12:12 h photoperiod, in a greenhouse. The highest nitrogen removal efficiencies (mainly NO3 -N and NO2 -N) and phosphorus (PO4 ) were observed in treatments with the highest proportion of D. asymmetricus, including T3 (25% C. vulgaris / 75 % D. asymmetricus), where 100.0% nitrogen and 77.1% phosphorus were removed. Likewise, T3 obtained the best result of dissolved oxygen (DO), achieving average of 25.50 ± 0.28 mg/L. Finally, according to the analysis of the mixture design, the optimal microalgae mixture was determinated that achieves the highest DO production, and the highest removal was the proportion of 6% of C. vulgaris and 94% of D. asymmetricus in 9 days cultivation time. In conclusion, microalgae have shown their capacity for bioremediation of domestic wastewater, which is an alternative to consider.

Selective activity of diselenides against Aedes aegypti (Diptera: Culicidae) larvae

Abstract INTRODUCTION: Aedes aegypti (L.) is the major vector of arboviruses that causes serious public health concerns in tropical and subtropical countries. METHODS: We examined the larvicidal activity of 1,2-diphenyldiselenide [(PhSe)2] and 1,2-bis(4-chlorophenyl) diselenide [(p-ClPhSe)2] and determine its toxicity to different non-target organisms. RESULTS: (PhSe)2 and (p-ClPhSe)2 killed Ae. aegypti L3 larvae with LC50/24h values of 65.63 µM (20.48 mg/L) and 355.19 µM (135.33 mg/L), respectively. (PhSe)2 was not toxic to the four model organisms. CONCLUSIONS: (PhSe)2 is a larvicidal compound with selective action against Ae. aegypti larvae. The mechanisms of action of (PhSe)2 under field conditions remain to be investigated.

Effect of the Application of Chlorella Vulgaris Ointment to the Number of Fibroblast Cells as an Indicator of Wound Healing in the Soft Tissue of Pig Ears

Abstract Objective: To analyze the clinical and histological condition of Chlorella vulgaris ointment extract smearing on wound healing. Material and Methods: The sample consisted of 9 pigs by making 4 incisions on the pig's right ear measuring 2cm x 1cm and applying Chlorella vulgaris ointment in 3 concentrations namely 5%, 10%, 15% and without ointment application as the control group. Furthermore, the observation of the clinical condition of wounds divided into 4 time periods (day 1, 3, 7 and 14). After completing clinical observations based on time period, the skin of the pig's ear was taken by cutting in the area that had been made injured. Skin tissue extraction carried out under inhalation and local anesthesia. The tissue pieces are then processed in the anatomical pathology laboratory to become 36 slide preparations. The clinical trial assessment was done using parameters of wound moisture, color of wound and scab wound. Whereas for histological examination was done by looking at the number of fibroblast cells in the CX31 light microscope with 40x magnification. ANOVA and Kruskal Wallis tests were used. Results: In the clinical observation conditions there were significant differences between the treatment group and the control group as well as the number of fibroblast cells there were significant differences between the 3 concentrations of Chlorella vulgaris extract ointment and the per time period control group. Conclusion: Chlorella vulgaris extract ointment containing 15% extract gives the best results in accelerating the wound healing process in the pig's ear seen from wound healing and increased number of fibroblast cells.

Effects of Sun and Oven Drying Techniques on Quality of Oil Produced from Chlorella vulgaris (Microalgae) Biomass

This study involved the molecular identification of Chlorella vulgaris microalgae obtained from Kaduna State University Fish Pond for biodiesel production potential. The DNA of Chlorella microalgae was extracted and subjected to PCR. The molecular weight of the PCR product obtained was 1.8kb using 18S rDNA primer sets and BLAST analyses revealed 95% identity with Chlorella vulgaris. The Chlorella vulgaris was cultured in open aquaria tanks at the Department of Biological Sciences, Nigerian Defence Academy. The biomass harvested was subjected to varying timings of sun and oven drying techniques (25-35ºC for 72hours and 60ºC for 12hours respectively) before extraction of oil from the biomass using solvent extraction method. The values for the density (0.854 and 0.867 cm3), specific gravity (0.875 and 0.876), acid value (0.414 and 0.384 mgKOH/g), saponification value (173.3 and 170.1 mgKOH/g), kinematic viscosity (5.200 and 3.870 mm2/g at 40ºC), flash point (114 and 115ºC) and cetane number (54.00 and 47.70) for the sun and oven dried biomass oil respectively were found to be in accordance with the ASTM standard values for biodiesel and fossil diesel. GC-MS analyses of the oil extracted using the two drying methods showed that the fatty acid profiling of the oil obtained from sun dried processed biomass had C14:0, C15:0, C16:0, C18:0, C18:1 cis9 and C22:1ω9 while the oven dried biomass oil had C14:0, C16:0, C19:0, C11:1, C18:1 cis9 and C22:1ω9. Drying methods, therefore, had influenced on the composition of saturated and unsaturated fatty acids. The oven dried biomass oil possesses high monounsaturated fatty acids when compared to sun dried biomass oil though the most important fatty acids (C14:0, C16:0 and C18:1) found in standard biodiesel were present in both. The results suggested that Chlorella vulgaris microalgae can be sustainably harvested for the production of biodiesel, both drying techniques can be employed for effective extraction but oven dried biomass oil was found to be of high quality because of the balanced in saturated and unsaturated fatty acid compositions and have an easy mode of operation but it required instrumentation.

Hematological and biochemical responses of pirarucu (Arapaima gigas, Arapaimidae) fed with diets containing a glucomannan product derived from yeast and algae / Respostas hematológicas e bioquímicas de pirarucu (Arapaima gigas, Arapaimidae) alimentado com dietas contendo mananoligossacarídeos derivados de leveduras e algas

ABSTRACT The hematological and biochemical responses of pirarucu fingerlings (Arapaima gigas) fed with diets containing different concentrations of a glucomannan product derived from yeast and algae were evaluated in order to ascertain the effect of these diets on fish physiology. Four treatments were conducted, with three replications, with 12 fish in each tank. The product evaluated (MycosorbA+(r)) was incorporated into the commercial diet, at four concentrations: 0, 1, 2 and 4 g.kg-1, called M0%, M0.1%, M0.2% and M0.4%, respectively. After 45 days of feeding, blood samples from six fish in each replicate were collected to perform the analyses. Their weight and length were determined to calculate the condition factor and weight gain, but no differences (P > 0.05) were observed among the treatments. No changes to the hematocrit, hemoglobin or erythrocyte levels or to the hematimetric indices of the pirarucus were observed. The glucose and triglyceride levels of the pirarucus in the M0.1% and M0.2% groups were significantly lower than those of the M0% group. The M0.2% group showed higher albumin levels (P < 0.05) than M0% and M0.4%. The M0.4% group showed a total cholesterol level that was significantly higher than in all other treatments. MycosorbA+(r) contributed towards increasing the levels of defense cells in A. gigas. It would be possible to use this product at concentrations of between 0.1% and 0.2%, given that they increase the levels of some defense cells and plasma albumin concentrations, without changes to hematological parameters, cholesterol and triglyceride plasma levels or condition factor.

RESUMO As características hematológicas e bioquímicas de alevinos de pirarucu Arapaima gigas alimentados com diferentes concentrações de mananoligossacarídeos derivados de leveduras e algas na dieta foram avaliadas para verificar o efeito sobre sua fisiologia. Quatro tratamentos foram conduzidos com três repetições (12 peixes em cada). O produto avaliado (MycosorbA+(r)) foi incorporado à ração comercial, em quatro níveis: 0, 1, 2 e 4 g kg-1 de ração, denominados M0%, M0,1%, M0,2% e M0,4%, respectivamente. Após 45 dias de alimentação, amostras de sangue de seis peixes de cada repetição foram coletadas para realização das análises. O peso e comprimento foram obtidos para cálculo do fator de condição e ganho de peso, entretanto, não foram observadas diferenças (P>0,05) entre os tratamentos. Assim como não foram observadas alterações nos valores de hematócrito, hemoglobina, eritrócitos e índices hematimétricos dos pirarucus. A concentração de glicose e triglicérides dos pirarucus dos grupos M0,1% e M0,2% foram significativamente menores que o tratamento M0%. Os peixes do tratamento M0,2% apresentaram níveis de albumina maior (P<0,05) que do M0% e M0,4%. Os peixes alimentados com M0,4% mostraram nível de colesterol total significativamente maior que todos os demais tratamentos. MycosorbA+(r) contribuiu para o aumento das células de defesa de A. gigas, podendo ser utilizado nas concentrações de 0,1% a 0,2%, devido ao aumento do número de certas células de defesa e dos níveis plasmáticos de albumina e por não ter sido observadas outras alterações nos parâmetros hematológicos, nos níveis plasmáticos de colesterol e triglicérides e no fator de condição.

INMOVILIZACIÓN DE LAS MICROALGAS Scenedesmus ovalternus (Scenedesmaceae) Y Chlorella vulgaris (Chlorellaceae) EN ESFERAS DE ALGINATO DE CALCIO / Immobilization of Microalgae Scenedesmus ovalternus (Scenedesmaceae) and Chlorella vulgaris (Chlorellaceae) in Calcium Alginate Beads

En este trabajo se describe la técnica de inmovilización de microalgas en esferas de alginato de calcio. Se emplearon las especies Scenedesmus ovalternus y Chlorella vulgaris, se determinó la estabilidad de las esferas, la cinética de crecimiento y la concentración de las microalgas en el interior de las esferas. Chlorella vulgaris alcanzó mayores densidades poblacionales y tasas de crecimiento más altas cuando se inmovilizó en concentraciones del 10 % v/v con el alginato (1,31*10(6) cél/ml). Para Scenedesmus ovalternus se observó una mayor densidad poblacional y una mayor tasa de crecimiento cuando se inmovilizó en concentraciones del 20 % v/v (7,06*10(5) cél/ml). Estos resultados son útiles para aplicaciones prácticas de las algas encapsuladas, tales como el biomonitoreo o la biorremediación.

This paper describes the immobilization technique of microalgae in calcium alginate beads. Scenedesmus ovalternus and Chlorella vulgaris species were used. The stability of beads, the kinetics of growth and the concentrations of microalgae inside the beads were determined. The higher density and the upper growth rate of Chlorella vulgaris occurred when it was immobilized in alginate at a concentration of 10 %v/v (1,31*10(6) cél/ml). Scenedesmus ovalternus achieved a higher population density and an elevated growth rate when it was immobilized at a concentration of 20 % v/v (7,06*10(5) cél/ml). These results are useful for subsequent applications of the encapsulated algae, such as biomonitoring and bioremediation.

Toxicological, phytochemical, and antibacterial assessment of Chlorella vulgaris and Spirulina platensis powder in albino rats. A preliminary study

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.

Potential role of marine algae extract on 3T3-L1 cell proliferation and differentiation: an in vitro approach

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.

Microalgae population dynamics in photobioreactors with secondary sewage effluent as culture medium

Nitrogen and phosphorus present in sewage can be used for microalgae growth, possibiliting cost reduction in the production of microalgae at the same time that it decreases the eutrophication potential of the effluent. This research aimed at monitoring the native community of microalgae and coliform bacteria in a secondary effluent from anaerobic municipal sewage treatment. Two treatments (aerated and non-aerated) were performed to grow microalgae under semi-controlled conditions in semi-closed photobioreactors in a greenhouse. The results showed no significant pH and coliforms (total and Escherichia coli) variation between treatments. Nutrient concentrations were reduced supporting microalgae growth up to 107 cells.mL−1 independent of aeration. Exponential growth was obtained from the first day for the non-aerated, but a 5 day lag phase of growth was obtained for the aerated. Chlorella vulgaris was the dominant microalgae (99.9%) in both treatments. In the aerated, 5 algae classes were detected (Chlorophyceae, Cyanophyceae, Chrysophyceae, Bacillariophyceae and Euglenophyceae), with 12 taxa, whereas in the non-aerated, 2 classes were identified (Chlorophyceae and Cyanophyceae), with 5 taxa. We concluded that effluent is viable for microalgae growth, especially Chlorella vulgaris, at the same time that the eutrophication potential and coliforms are decreased, contributing for better quality of the final effluent.

Eco-Friendly Approach for Treating Dairy Effluent and Lipid Estimation Using Microalgae.

Aim: In this study, an attempt has been made to examine the utility of these species in treating dairy wastewater. Bioreactor was studied using immobilized Chlorella vulgaris and Anabaena ambigua to treat dairy effluent. Study Design: The entire study including the treatment and filtration was conducted in Centre for biotechnology, Institute of Science and Technology, Jawaharlal Nehru Technological University, Hyderabad, between November 2014 to February 2015. Methodology: The treatment of dairy effluent consists of two stages; the first stage includes dairy effluent treatment using immobilised Chlorella vulgaris and Anabeana ambigua, while the second stage involves sand bed and coal bed filtration. Results: Chlorella vulgaris reported a high lipid content of 12% when compared with Anabaena ambigua 5% after the cultivation period. Whereas the protein content of Anabaena ambigua (40%) was higher than Chlorella vulgaris (28%) when compared with after harvesting. Whilst ammonium nitrate was completely removed by bead treatment it was 96% reduction when treated with Chlorella vulgaris. A 98% removal of phosphates was achieved on an average after algal bead treatment for both species in both modes of operation. Moreover, a significant decrease in BOD and COD was achieved by this method. Conclusion: It can be concluded that, the cultivation of algae in dairy effluent results the combined advantages of treating the dairy effluent and also producing algal biomass, which can further use of food additives for aquatic culture, animal and human feed, energies such as biogas and fuels and bio-fertiliser.

Efecto del alimento sobre variables productivas de dos especies de Cladóceros bajo condiciones de laboratorio / Effect of food intake on productive variables of two species of cladocera, under laboratory conditions

Objetivo. Evaluar los efectos de diferentes tipos de alimentos sobre la productividad de Diaphanosoma sp., y Alona sp bajo condiciones de laboratorio. Materiales y métodos. Juveniles de Diaphanosoma sp., y Alona sp., fueron sembrados en acuarios de 20 L a una densidad de 6 org L-1 y mantenidos con aireación constante y fotoperíodo 12:12 (luz : oscuridad). Se alimentaron con Saccharomyces cereviseae y Chlorella vulgaris a concentración de 4 x 10(5) Cel/ml cada uno, en tres combinaciones: 1. C. vulgaris, 2. S. cereviseae, 3. C. vulgaris + S. cereviseae en proporciones iguales; de cada tratamiento se realizaron 3 réplicas. Con el propósito de determinar la densidad poblacional, cada 3 días y durante un período de 27 días, y se midió la concentración de individuos por ml. Resultados. Se obtuvieron mejores resultados en el tratamientos de microalgas + levadura, con valores de TCE=0.1323±0.07 y 0.1598±0.04 días-1; R=0.010 ±0.004 y 0.019±0.01 org ml-1 día-1, TD=5.2±0.5 y 4.3±0.38 días para Diaphanosoma sp., y Alona sp., respectivamente, pero sin presentar diferencias significativas (p>0.05). Conclusiones. La dieta mixta de levadura y microalgas produjo mayor eficiencia productiva en las dos especies estudiadas, constituyendo una alternativa de fuente de alimento vivo para la acuicultura.

Objective. Assess the effects of different types of food intake on the productivity of Diaphanosoma sp. and Alona sp. under laboratory conditions. Materials and methods. Young Diaphanosoma sp. and Alona sp. were planted in 20 L aquariums at a density of 6 org L-1 and maintained with constant aeration and a photoperiod 12:12 (light: dark). The organisms were fed with Saccharomyces cereviseae and Chlorella vulgaris at a concentration of 4 x 10(5) cells/ml in three combinations: 1. C. vulgaris, 2. S. cereviseae 3. C. vulgaris + S. cereviseae); 3 replicas were carried out for each treatment. To determine the population density, every 3 days over a 27 day period, the concentration of individuals per ml was measured using a Bogorov camera. Results. The best results were obtained in treatments with microalgae and yeast, with values of TCE=0.1323±0.07 and 0.1598±0.04 days-1, R=0.010±0.004 and 0.019±0.01 org ml-1 days-1, TD=5.2±0.5 and 4.3±0.38 days for Diaphanosoma sp., and Alona sp., respectively, but without significant differences (p>0.05). Conclusions. The mixed diet of yeast and microalgae produced greater productive efficiency in the studied species, constituting an alternative source of live feed for aquaculture.

Gene transfer of Chlorella vulgaris n-3 fatty acid desaturase optimizes the fatty acid composition of human breast cancer cells

Chlorella vulgaris has the gene of n-3 fatty acid desaturase (CvFad3), which can synthesize the precursor of n-3 polyunsaturated fatty acids (PUFAs) or convert n-6 to n-3 PUFAs. The objective of the present study was to examine whether the CvFad3 gene from C. vulgaris can be functionally and efficiently expressed in human breast cancer cells and whether its expression can exert a significant effect on cell fatty acid composition. We inserted the CvFad3 gene into the plasmid pEGFP-C3 to construct the eukaryotic expression vector pEGFP-C3-n-3 and to express the n-3 Fad gene in human breast cancer cells (MCF-7 cells). Transfection of MCF-7 cells with the recombinant vector resulted in a high expression of n-3 fatty acid desaturase. Lipid analysis indicated that the ratio of n-6/n-3 PUFAs was decreased from 6:1 in the control cells to about 1:1 in the cells expressing the n-3 fatty acid desaturase. Accordingly, the CvFad3 gene significantly decreased the ratio of n-6/n-3 PUFAs of the MCF-7 cell membrane. The expression of the CvFad3 gene can decrease cell proliferation and promote cell apoptosis. This study demonstrates that the CvFad3 gene can dramatically balance the ratio of n-6/n-3 PUFAs and may provide an effective approach to the modification of the fatty acid composition of mammalian cells, also providing a basis for potential applications of its transfer in experimental and clinical settings.

Antioxidant enzyme activity and malondialdehyde levels can be modulated by Piper betle, tocotrienol rich fraction and Chlorella vulgaris in aging C57BL/6 mice

OBJECTIVE: The aim of this study was to determine the erythrocyte antioxidant enzyme activity and the superoxide dismutase, catalase, glutathione peroxidase, and plasma malondialdehyde levels in aging mice and to evaluate how these measures are modulated by potential antioxidants, including the tocotrienol-rich fraction, Piper betle, and Chlorella vulgaris. METHOD: One hundred and twenty male C57BL/6 inbred mice were divided into three age groups: young (6 months old), middle-aged (12 months old), and old (18 months old). Each age group consisted of two control groups (distilled water and olive oil) and three treatment groups: Piper betle (50 mg/kg body weight), tocotrienol-rich fraction (30 mg/kg), and Chlorella vulgaris (50 mg/kg). The duration of treatment for all three age groups was two months. Blood was withdrawn from the orbital sinus to determine the antioxidant enzyme activity and the malondialdehyde level. RESULTS: Piper betle increased the activities of catalase, glutathione peroxidase, and superoxide dismutase in the young, middle, and old age groups, respectively, when compared to control. The tocotrienol-rich fraction decreased the superoxide dismutase activity in the middle and the old age groups but had no effect on catalase or glutathione peroxidase activity for all age groups. Chlorella vulgaris had no effect on superoxide dismutase activity for all age groups but increased glutathione peroxidase and decreased catalase activity in the middle and the young age groups, respectively. Chlorella vulgaris reduced lipid peroxidation (malondialdehyde levels) in all age groups, but no significant changes were observed with the tocotrienol-rich fraction and the Piper betle treatments. CONCLUSION: We found equivocal age-related changes in erythrocyte antioxidant enzyme activity when mice were treated with Piper betle, the tocotrienol-rich fraction, and Chlorella vulgaris. However, Piper betle treatment showed increased antioxidant enzymes activity during aging.

Population dynamics of the tropical cladoceran Ceriodaphnia rigaudi Richard, 1894 (Crustacea: Anomopoda). Effect of food type and temperature.

The knowledge of population effects of food on tropical, filter-feeding cladocerans is scarce because a reduced number of species has been extensively studied. Ceriodaphnia rigaudi Richard 1894, a small-sized cladoceran distributed mainly in tropical and subtropical regions of the world, was studied. The aim of this study was to contribute to the knowledge of the reproductive biology of a poor-known Cladoceran; for this we assessed the effect of feeding and temperature on the reproduction and life cycle of this species. Three microalga species (Pseudokirchneriella subcapitata, Ankistrodesmus falcatus, and Chlorella vulgaris) were supplied as food each at a concentration of 12 mg l-1 (dry weight, equivalent to 1.3X106, 0.4X106 and 1.35X106 cell ml-1, respectively, and equivalent to 7.80 Eg C ml-1), at two temperatures (20 and 25oC). We evaluated, among other responses, longevity, total progeny, survival, life expectancy at birth and fecundity. Organisms fed with the microalgae A. falcatus and P. subcapitata presented both higher longevity (30.7± 5.91, 26.6 ± 3.59 days, respectively) and total progeny (45 ± 13.80, 40.7 ± 0.66 neonates female-1) values than those organisms fed C. vulgaris (13.5± 4.63 days and 17.6 ± 6.19 neonates female-1, respectively). On the other hand, temperature affected significantly the population parameters of C. rigaudi, recording maximal longevity values (56.1 ± 9.41 days) at 20oC in organisms fed A. falcatus; however, age at first reproduction and total progeny were negatively affected by this temperature: sexual maturation of the females was delayed until the age of 16 days and the number of neonates produced was smaller (9.8 ± 3.45 with C. vulgaris; 24.7±6.01 with P. subcapitata, and 35.5 ± 8.59 neonates female-1 with A. falcatus). The best reproductive responses for C. rigaudi in this study were obtained with A. falcatus at 25oC.

Cultivation of algae in photobioreator and obtention of biodiesel

In this work we described the cultivation of Chlorella vulgaris in a photobioreactor to algal biomass production. The dried biomass was used as feedstock for biodiesel production, it presented 26 percent lipids and via sonocatalysis stage of the methodology resulted in 60 percent of fatty acid methyl esters (FAME). The FAME content was confirmed by Gas Chromatography (GC).

Does Chlorella vulgaris Modulate the Expression of COL and MMP Genes in Skin Ageing?

Chlorella vulgaris, a unicellular microalgae, produces many intracellular phytochemicals namely carotenoids, tocopherols, ubiquinone and protein. Skin ageing which is induced by oxidative stress involves decreased extracellular matrix synthesis and increased expression of enzymes that degrade the collagenous matrix. The objective of this study was to determine the effect of C. vulgaris on the expression of genes encoded for collagen (COL) and matrix metalloproteinases (MMPs) which are involved in skin ageing. Human diploid fibroblasts (HDFs) were obtained from circumcision foreskin of 8-12 year-old boys. HDFs were cultured into 3 groups: untreated control cells, cells with stress-induced premature senescence (SIPS; cells were induced with H2O2 at passage 6 for 2 weeks) and SIPS treated with C. vulgaris (prolonged C. vulgaris treatment started at passage 4 and combined treatment with H2O2 at passage 6 for 2 weeks). Senescence-associated ß-galactosidase (SA ß-gal) was determined using senescent cells histochemical staining kit (Sigma, USA). Expression of COLI, COLIII, COLIV, MMPI, MMPII and MMPIII genes was quantitatively analysed with real-time RT-PCR method (iScript™ One Step real-time PCR with SYBR® Green; Biorad). HDFs treated with H2O2 (SIPS) exhibited senescent morphological features of flattening and enlarged with increased expression of SA ß-gal (p<0.05). Gene expression analysis showed COLI was downregulated in SIPS and SIPS treated with C. vulgaris (p<0.05) while COLIII decreased in SIPS and increased in SIPS treated with C. vulgaris (p<0.05). Expression of MMPI was increased in SIPS and SIPS treated with C. vulgaris (p<0.05) indicating its synergistic effect with H2O2 treatment. In conclusion, in skin ageing, COLI and COLIII genes were downregulated while MMPI was upregulated. C. vulgaris modulated the expression of COL and MMP genes by downregulating COLI and upregulating COLIII and MMPI but it did not exert anti-ageing effect.

Hot water extract of Chlorella vulgaris induced DNA damage and apoptosis

OBJECTIVES: The aim of this study was to determine the antiproliferative and apoptotic effects of hot water extracts of Chlorella vulgaris on hepatoma cell line HepG2. INTRODUCTION: The search for food and spices that can induce apoptosis in cancer cells has been a major study interest in the last decade. Chlorella vulgaris, a unicellular green algae, has been reported to have antioxidant and anti-cancer properties. However, its chemopreventive effects in inhibiting the growth of cancer cells have not been studied in great detail. METHODS: HepG2 liver cancer cells and WRL68 normal liver cells were treated with various concentrations (0-4 mg/ml) of hot water extract of C. vulgaris after 24 hours incubation. Apoptosis rate was evaluated by TUNEL assay while DNA damage was assessed by Comet assay. Apoptosis proteins were evaluated by Western blot analysis. RESULTS: Chlorella vulgaris decreased the number of viable HepG2 cells in a dose dependent manner (p < 0.05), with an IC50 of 1.6 mg/ml. DNA damage as measured by Comet assay was increased in HepG2 cells at all concentrations of Chlorella vulgaris tested. Evaluation of apoptosis by TUNEL assay showed that Chlorella vulgaris induced a higher apoptotic rate (70 percent) in HepG2 cells compared to normal liver cells, WRL68 (15 percent). Western blot analysis showed increased expression of pro-apoptotic proteins P53, Bax and caspase-3 in the HepG2 cells compared to normal liver cells WRL68, and decreased expression of the anti-apoptotic protein Bcl-2. CONCLUSIONS: Chlorella vulgaris may have anti-cancer effects by inducing apoptosis signaling cascades via an increased expression of P53, Bax and caspase-3 proteins and through a reduction of Bcl-2 protein, which subsequently lead to increased DNA damage and apoptosis.

Accumulation of chromium and interaction with other elements in Chlorella vulgaris (Cloroficeae) and Daphnia magna (Crustacea, Cladocera).

Assays with Chlorella vulgaris Beijerinck Novakova, 1890 and Daphnia magna Straus, 1820 were performed to determine Cr and other elements concentration in tissues of both species by instrumental neutron activation analysis (INAA), after being exposed to 150, 280 and 350 .g l-1Cr (VI). Interaction among Cu, Zn, Fe and Cr were also registered. In the control of C. vulgaris, the amount of Cr was < 4μg g-1; in the treatments with Cr (VI) the values were 47, 82 and 100 folds greater than the control for the lowest, intermediate and highest concentrations tested respectively. In the control of D. magna, the amount of Cr was <3 μg g-1; in the treatments with Cr (VI) the values were 14, 13 and 27 folds higher than the control for the lower, intermediate and higher Cr (VI) concentrations respectively, and from 3 to 9 times less than for C. vulgaris These results show that C. vulgaris is very efficient accumulator of Cr (VI) from polluted waters, and in consequence, it is proposed to be used in phytoremediation procedures.

Effect of Chlorella vulgaris intake on cadmium detoxification in rats fed cadmium

The aim of this study was to investigate if dietary Chlorella vulgaris (chlorella) intake would be effective on cadmium (Cd) detoxification in rats fed dietary Cd. Fourteen-week old male Sprague-Dawley (SD) rats weighing 415.0 +/- 1.6 g were randomly divided into two groups and fed slightly modified American Institute of Nutrition-93 Growing (AIN-93G) diet without (n=10) or with (n=40) dietary Cd (200 ppm) for 8 weeks. To confirm alteration by dietary Cd intake, twenty rats fed AIN-93G diet without (n=10) and with (n=10) dietary Cd were sacrificed and compared. Other thirty rats were randomly blocked into three groups and fed slightly modified AIN-93G diets replacing 0 (n=10), 5 (n=10) or 10% (n=10) chlorella of total kg diet for 4 weeks. Daily food intake, body weight change, body weight gain/calorie intake, organ weight (liver, spleen, and kidney), perirenal fat pad and epididymal fat pad weights were measured. To examine Cd detoxification, urinary Cd excretion and metallothonein (MT) concentrations in kidney and intestine were measured. Food intake, calorie intake, body weight change, body weight gain/calorie intake, organ weight and fat pad weights were decreased by dietary Cd intake. Urinary Cd excretion and MT concentrations in kidney and small intestine were increased by dietary Cd. After given Cd containing diet, food intake, calorie intake, body weight change, body weight gain/calorie intake, organ weights and fat pad weights were not influenced by dietary chlorella intake. Renal MT synthesis tended to be higher in a dose-dependent manner, but not significantly. And chlorella intake did not significantly facilitate renal and intestinal MT synthesis and urinary Cd excretion. These findings suggest that, after stopping cadmium supply, chlorella supplementation, regardless of its percentage, might not improve cadmium detoxification from the body in growing rats.

Effect of Chlorella vulgaris on lipid metabolism in Wistar rats fed high fat diet

This study was performed to investigate effects of Chlorella vulgaris on lipid metabolism in rats fed high fat diet. Sixty 6-week-old male Wistar rats were divided into two groups; normal diet group and high fat diet group, then the rats in each group were further divided into three subgroups and fed 0%, 5% and 10% (w/w) chlorella-containing diets, respectively, and raised for 9 weeks. Serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT) activity and total protein and albumin concentration were not different among groups. Serum total lipids and liver TG concentration were significantly lower in 5% and 10% chlorella groups than 0% chlorella group in high fat diet groups (p<0.05). Serum TG, serum total cholesterol, liver total lipid and liver total cholesterol concentrations were significantly lower in 10% chlorella groups than 0% chlorella group in high fat diet groups (p<0.05). Fecal total lipid, TG and total cholesterol excretions were significantly higher in 5% and 10% chlorella groups than 0% chlorella groups in normal diet and high fat diet groups, respectively (p<0.05). These results suggest that Chlorella vulgaris is effective for prevention of dyslipidemia which may be due to the modulation of lipid metabolism and increased fecal excretion of lipid.