The aim of this study is to produce a pharmaceutical grade single cell product of Arthrospira from a mixed culture. We have designed a medium derived from a combination between George’s and Zarrouk’s media. Our new medium has the ability to inhibit different forms of cyanobacterium and microalgae except the Chlorella. The medium and the cultivation conditions have been investigated to map the points where only Arthrospira could survive. For that, a mixed culture of pure Chlorella and Arthrospira (~90？:？10) has been used to develop the best medium composition that can lead to the enrichment of the Arthrospira growth and the inhibition of the Chlorella growth. To enable better control and to study its growth, an 80 l photobioreactor has been used. We have used high saline (2xA-St) medium which has been followed by in fermentor reducing its concentration to 1.5x. The investigation proves that Chlorella has completely disappeared. A method and a new saline medium have been established using a photobioreactor for in fermentor production of single cell Arthrospira. Such method enables the production of pure pharmaceutical grade Arthrospira for medicinal and pharmaceutical applications or as a single cell protein. 1. Introduction Using algae as a food and medicine is deeply rooted in the human history. In ancient Egypt and today, farmers used to collect floating algae on the surface of the water to feed their domestic birds. In a harsh environment when the land resources food become rare, alkaline lakes play a significant role as an alternative source. The alkaline lakes enable the growth of one of the few nontoxic cyanobacterial species, Arthrospira fusiformis [1]. Humans learned early how to use the Arthrospira as a food source. Seeing the migrant birds feed safely on Arthrospira, such as lesser flamingoes (Phoeniconaias minor Geoffroy), encourages such use. Kebeda (1997) reported that in Ethiopia, farmers and herdsmen living in areas close to the soda lakes make their cattle drink Arthrospira water about once a month and believe that it has therapeutic effects and compensates for some lack in dietary food [2]. The invention of the microscope enabled Turpin in 1827 to identify and describe Arthrospira as spiral cyanobacteria [3]. Species of Arthrospira have been found in a variety of environments including soil, sand, marshes, brackish water, seawater, and freshwater [1, 4]. Rich (1931) has reported it as a dominant phytoplankton in a number of lakes in the Rift Valley of East Africa [5]. 113 years after its first microscopic identification, Arthrospira was
References
[1]
O. Ciferri, “Spirulina, the edible microorganism,” Microbiological Reviews, vol. 47, no. 4, pp. 551–578, 1983.
[2]
E. Kebede, “Response of Spirulina platensis (=Arthrospira fusiformis) from Lake Chitu, Ethiopia, to salinity stress from sodium salts,” Journal of Applied Phycology, vol. 9, no. 6, pp. 551–558, 1997.
[3]
P. J. F. Turpin, “Spirulina oscillarioide,” in Dictionnaire des Sciences Naturelles, vol. 50, pp. 309–310, DeLevrault, Paris, France, 1827.
[4]
G.-G. Choi, C.-Y. Ahn, and H.-M. Oh, “Phylogenetic relationships of Arthrospira strainsinferred from16SrRNA geneand cpcBA-IGS sequences,” Algae, vol. 27, no. 2, pp. 75–82, 2012.
[5]
F. Rich, “Notes on Arthrospira platetnsis,” Rev Algol, vol. 6, pp. 75–79, 1931.
[6]
P. Dangeard, “Sur une algue bleue alimentaire pour l'homme: Arthrospira platensis (Nordst.),” Gomont. Actes Soc. Linn. Boreaux Extr. Procés-Verbaux, vol. 91, pp. 39–41, 1940.
[7]
G. Chamorro, M. Salazar, K. G. de Lima Araújo, C. P. dos Santos, G. Ceballos, and L. F. Castillo, “Update on the pharmacology of Spirulina (Arthrospira), an unconventional food,” Archivos Latinoamericanos de Nutricion, vol. 52, no. 3, pp. 232–240, 2002.
[8]
A. Richmond, “Large scale microalgal culture and applications,” in Progressin Phycological Research, vol. 7, Biopress, Bristol, UK, 1990.
[9]
I. Avila-Leon, M. Chuei Matsudo, S. Sato, and J. C. M. De Carvalho, “Arthrospira platensis biomass with high protein content cultivated in continuous process using urea as nitrogen source,” Journal of Applied Microbiology, vol. 112, pp. 1086–1094, 2012.
[10]
H. Shimamatsu, “Mass production of Spirulina, an edible microalga,” Hydrobiologia, vol. 512, pp. 39–44, 2004.
[11]
C. Zarrouk, Contribution a l’etude d’une cyanophycee. Influence de divers facteurs physiques et chimiques sur la croissance et la photosynthese de Spirulina maxima (Setch. et Gardner). Geitler [Ph.D. thesis], University of Paris, Paris, France, 1966.
[12]
A. Iltis, “Tolerance de salinite de Spirulina platensis (Gom.). Geitl., (Cyanophyta). dans les mares natronees du Kanem (Tchad),” Cahiers ORSTOM: Série Hydrobiologie, vol. 2, pp. 119–125, 1968.
[13]
E. A. Davis, “Growth of algae in mass culture: plastic and glass tubing,” in Algal Culture: From LaboraTory To Pilot Plant, J. S. Burlew, Ed., pp. 113–153, Carnegie Institution of Washington, Washington, DC, USA, 1953.
[14]
Y.-K. Lee and C.-S. Low, “Effect of photobioreactor inclination on the biomass productivity of an outdoor algal culture,” Biotechnology and Bioengineering, vol. 38, no. 9, pp. 995–1000, 1991.
[15]
O. Pulz and P. Kretschmer, “Perspectives of phototrophic microorganisms in environment protection and ecology,” Acta Biotechnologica, vol. 12, no. 6, pp. 517–520, 1992.
[16]
T. Hai, H. Ahlers, V. Gorenflo, and A. Steinbüchel, “Axenic cultivation of anoxygenic phototrophic bacteria, cyanobacteria, and microalgae in a new closed tubular glass photobioreactor,” Applied Microbiology and Biotechnology, vol. 53, no. 4, pp. 383–389, 2000.
[17]
S. Oncel and F. V. Sukan, “Comparison of two different pneumatically mixed column photobioreactors for the cultivation of Artrospira platensis (Spirulina platensis),” Bioresource Technology, vol. 99, no. 11, pp. 4755–4760, 2008.
[18]
M. C. Matsudo, R. P. Bezerra, S. Sato, A. Converti, and J. C. M. de Carvalho, “Photosynthetic efficiency and rate of CO2 assimilation by Arthrospira (Spirulina) platensis continuously cultivated in a tubular photobioreactor,” Biotechnology Journal, vol. 7, pp. 1412–1417, 2012.
[19]
D. L. Narasimha, G. S. Venkataraman, S. K. Duggal, and B. O. Eggum, “Nutritional quality of the blue-green alga Spirulina platensis Geitler,” Journal of the Science of Food and Agriculture, vol. 33, no. 5, pp. 456–460, 1982.
[20]
A. Belay, T. Kato, and Y. Ota, “Spirulina (Arthrospira): potential application as an animal feed supplement,” Journal of Applied Phycology, vol. 8, no. 4-5, pp. 303–311, 1996.
[21]
J. J. Ortega-Calvo, C. Mazuelos, B. Hermosin, and C. Saiz-Jimenez, “Chemical composition of Spirulina and eukaryotic algae food products marketed in Spain,” Journal of Applied Phycology, vol. 5, no. 4, pp. 425–435, 1993.
[22]
T. Hirata, M. Tanaka, M. Ooike, T. Tsunomura, and M. Sakaguchi, “Antioxidant activities of phycocyanobilin prepared from Spirulina platensis,” Journal of Applied Phycology, vol. 12, no. 3-5, pp. 435–439, 2000.
[23]
T. Kato, “Blue pigment from Spirulina,” New Food Industry, vol. 29, pp. 17–21, 1994.
[24]
A. T. Kursar and S. R. Alberte, “Photosynthetic unit organization in a red algae,” Plant Physiology, vol. 72, pp. 409–414, 1983.
[25]
R. Rodríguez-Sánchez, R. Ortiz- Butrón, V. Blas-Valdivia, A. Hernández-García, and E. Cano-Europa, “Phycobiliproteins or C-phycocyanin of Arthrospira (Spirulina) maxima protect against HgCl(2)-caused oxidative stress and renal damage,” Food Chemistry, vol. 135, no. 4, pp. 2359–2365, 2012.
[26]
M. Sharaf, A. Amara, A. Aboul-Enein et al., “Molecular authentication and characterization of the antiherpetic activity of the cyanobacterium Arthrospira fusiformis,” Pharmazie, vol. 65, no. 2, pp. 132–136, 2010.
[27]
T. Hayashi, K. Hayashi, M. Maeda, and I. Kojima, “Calcium spirulan, an inhibitor of enveloped virus replication, from a blue-green alga Spirulina platensis,” Journal of Natural Products, vol. 59, no. 1, pp. 83–87, 1996.
[28]
H. Gong, Y. Tang, J. Wang, X. Wen, L. Zhang, and C. Lu, “Characterization of photosystem II in salt-stressed cyanobacterial Spirulina platensis cells,” Biochimica et Biophysica Acta, vol. 1777, no. 6, pp. 488–495, 2008.
[29]
C. Lu and J. Zhang, “Role of light in the response of PSII photochemistry to salt stress in the cyanobacterium Spirulina platensis,” Journal of Experimental Botany, vol. 51, no. 346, pp. 911–917, 2000.
[30]
R. M. Atlas and L. C. Parks, Handbook of Microbiological Media, CRC. Press, Boca Raton, Fla, USA, 2nd edition, 1997.
[31]
H. Chen and S.-S. Pan, “Bioremediation potential of spirulina: toxicity and biosorption studies of lead,” Journal of Zhejiang University, vol. 6, no. 3, pp. 171–174, 2005.
[32]
A. V. Kotinsky, L. A. Chernukhina, G. V. Donchenko et al., “Effect of iodine and cobalt salts on biologically active substances content in spirulina biomass,” Ukrain'skyi Biokhimichnyi Zhurnal, vol. 76, no. 2, pp. 112–116, 2004.
[33]
A. Lodi, L. Binaghi, C. Solisio, A. Converti, and M. Del Borghi, “Nitrate and phosphate removal by Spirulina platensis,” Journal of Industrial Microbiology and Biotechnology, vol. 30, no. 11, pp. 656–660, 2003.
[34]
M. Ruengjitchatchawalya, N. Chirasuwan, R. Chaiklahan, B. Bunnag, M. Tanticharoen, and P. Deshnium, “Photosynthetic characterization of a mutant of Spirulina platensis,” Journal of Applied Phycology, vol. 14, no. 2, pp. 71–76, 2002.
