CULTIVATION OF Chlorella sorokiniana IN ALTERNATIVE MEDIA WITH AGRICULTURAL FERTILIZER AND VINASSE

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1 CULTIVATION OF Chlorella sorokiniana IN ALTERNATIVE MEDIA WITH AGRICULTURAL FERTILIZER AND VINASSE Carvalho, E.M. 1 ; Pereira, N. S. 1 ; Ribeiro, D.M. 2 ; Fonseca, G. G. 2 ; and Menegazzo M.L. 2 1 Faculty of Biological and Environmental Sciences (FCBA), Federal University of Grande Dourados (UFGD), Dourados-MS, Brazil, ( emersoncarvalho@ufgd.edu.br, nathaskia.spn@outlook.com) 2 Faculty of Engineering (FAEN), Federal University of Grande Dourados (UFGD), Dourados-MS, Brazil, ( dagonribeiro@hotmail.com, ggf@ufgd.edu.br, marianamenegazzo@ufgd.edu.br)

2 CULTIVATION OF Chlorella sorokiniana IN ALTERNATIVE MEDIA WITH AGRICULTURAL FERTILIZER AND VINASSE Carvalho, E.M. 1 ; Pereira, N. S. 1 ; Ribeiro, D.M. 2 ; Fonseca, G. G. 2 ; and Menegazzo M.L. 2 1 Faculty of Biological and Environmental Sciences (FCBA), Federal University of Grande Dourados (UFGD), Dourados-MS, Brazil, ( emersoncarvalho@ufgd.edu.br, nathaskia.spn@outlook.com) 2 Faculty of Engineering (FAEN), Federal University of Grande Dourados (UFGD), Dourados-MS, Brazil, ( dagonribeiro@hotmail.com, ggf@ufgd.edu.br, marianamenegazzo@ufgd.edu.br) Abstract New alternative culture media are tested in a bench bioassay for the production of microalgae in order to make them economically viable at industrial scale. Thus, the objective of this study was to evaluate the growth of microalgae Chlorella sorokiniana in alternative media enriched with NPK and vinasse. The microalgae were cultured in axenic not static system with constant aeration at room temperature and photoperiod of 2000 lux (12 hours light / 12 h dark) and its density evaluated with vinasse concentration of 0.1%, 1% and 10%, and agricultural fertilizer N: P: K ( g.l -1 ) concentration of 1%. It was observed that the treatments with vinasse 1 and 10% were about two times larger than the others. However, the vinasse presented a great potential to be used as a culture medium in order to produce algal biomass with high added value. This is because the tests containing higher concentrations of this residue were the ones with the best growth parameters. Keywords Bioassay; chlorophyceae; Environmental biotechnology; Microalgae. INTRODUCTION Microalgae are unicellular organisms capable of photosynthesis faster and more efficiently than land plants (Carvalho et al., 2012). These microorganisms are a group of heterogeneous organism and generally microscopic unicellular colored due to the presence of photosynthetic pigments (chlorophyll) occur primarily in water, photoautotrophic (not necessarily all the time), and may be prokaryotic or eukaryotic (Olaizola, 2003). New alternative culture media are tested in a bench bioassay for the production of microalgae in order to make them economically viable at industrial scale (Sipaúba- Tavares et al., 2009; Carvalho et al., 2012). Thus, various types of industrial wastes are being tested as a source of nutrients for the cultivation of microalgae, for example, glycerol (by-product of biodiesel production), the animal manure and vinasse (Pyle et al., 2008; Fioresi and Sipaúba Tavares, 2008; Kobayashi et al., 2013; Barrocal et al., 2010). These wastes, however, have inherent characteristics that give it the potential of microalgae as a culture medium. The vinasse, for example, contains more than 50% of sugars, and other nutrients, such as N and K, enabling its use as a carbon source for the supplementation of microalgae cultivation (Andrade and Costa, 2008). Thus, its adoption may lower the costs of large-scale production and serve as a solution to the reutilization of highly polluting waste. Given these arguments, the objective of this study was to evaluate

3 the growth of microalgae Chlorella sorokiniana in alternative media enriched with NPK and vinasse. MATERIAL AND METHODS The strain of C. sorokiniana CTT 7727 was provided by André Tosello Institute (Ref ). To evaluate the kinetics of growth of microalgae cultivation media were supplemented with vinasse - concentration of 0.1%, 1% and 10% (v/v) - and agricultural fertilizer N: P: K -1% concentration solution of stock (v/v) - all autoclaved at 121 C for 20 minutes. The stock solution was prepared by adding 0.70 g of chemical fertilizer N:P:K (20/05/20 g.l -1 ) in ml distilled water (Sipaúba- Tavares and Rocha, 2003). NPK Vinasse 0.1% Vinasse 1% Vinasse 10% microalgae cultivation in plastic bag fluorescent lighting aeration hose Figure 1. Structure of the experimental design for the production of microalgae. The management system used was not static axenic, with constant aeration, ambient temperature and photoperiod 2000 LUX (12 h light / 12 h dark). The cultures were conditioned in plastic bags containing the total volume of 1000 ml (Figure 1). Cell density was monitored every five days through the chamber Neubauer counting for obtaining algal density. RESULTS AND DISCUSSION Mean values of cell density obtained in the bioassay are shown in Figure 2, where it can be seen a continuous growth of microalgae C. sorokiniana in all treatments during the 30 days. However, note that the treatments with vinasse 1 and 10% were about two times larger than the others, with values of 131 and x 10 5 cells/ml, compared with values of 55.5 and 58.2 x 10 5 cells/ml in treatments with vinasse 0.1% and NPK. These differences indicate that the microalgae C. sorokiniana showed higher productive efficiency and biomass conversion in the media with higher concentrations of the residue.

4 160 Vinasse 0,1% Vinasse 1% Vinasse 10% NPK Cell density (x 10 5 cells.ml -1 ) Time (days) Figure 2. Cell density (mean ± standard error) of microalgae C. sorokiniana grown in medium supplemented with NPK fertilizer and vinasse. Therefore, we observed that the vinasse did not show any significant degree of toxicity for microalgae C. sorokiniana the concentrations studied. Kadioǧlu and Algur (1992) observed a significant decrease in the number of microalgae Chlamydomonas reinhardii cells in media with vinasse in concentrations above 10%, attributed this decrease to a possible osmotic effect due to the high content of organic matter present in the medium, and also by the likely presence of toxic elements such as Zn, Mn and Fe. There were no deleterious effects on C. sorokiniana produced in vinasse 10%. This indicates a high potential of microalgae in the treatment of this waste. In addition, microalgae can present high capacity to remove nutrients like N, P and organic matter in different conditions, and wastewater (Kim et al., 2013;Liu et al., 2012). CONCLUSION However, the vinasse presented a great potential to be used as a culture medium in order to produce algal biomass with high added value. This is because the tests containing higher concentrations of this residue were the ones with the best growth parameters. REFERENCES Andrade, M.D.R.; Costa, J.A.V Culture of microalga Spirulina platensis in alternative sources of nutrients. Ciência e agrotecnologia, 32, Barrocal, V. M. et al., 2010 Production of biomass by Spirulina maxima using sugar beet vinasse in growth media. New biotechnology, 27, Carvalho, E.M., et al., 2012 Growth kinetics of the microalga Pseudokirchneriella subcapitata (Korshikov) Hindak (Chlorophyceae) in natural water enriched with NPK fertilizer. Biochemistry And Biotechnology Reports, 01, Fioresi, T.B.; Sipaúba-Tavares, L.H Cultivo de Ankistrodesmus gracilis (Chlorophyta) em laboratório à base de esterco suíno. Revista Biotemas, 21, Kadioǧlu, A.; Algur, Ö.F Tests of media with vinasse for Chlamydomonas reinhardii for possible reduction in vinasse pollution. Bioresource technology, 42, 1-5.

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