In-Door Mass Culture of Nannochloropsis oculata

Preparation of materials Materials used were carboys, glass tubings, air hose, and pipettes. The trainee washed the materials with detergents and then soaked them in water containing chlorine for few minutes and then air dried. Glass tubings and pipettes was wrapped with paper or aluminium foil and then oven dried at 1500C for 30 to 45 minutes.

Preparation of seawater in carboys Seawater was pumped to a 1 toner white reservoir using filter bag with .05 microns and was transferred to a 1 toner black reservoir container passing through .05 microns cartridge filter. The seawater was then added with 400 ml chlorine and aerated for few minutes. The chlorinated seawater could only be used the next day. The trainee filled 7 liters chlorinated seawater in each carboy (Appendix C, Plate 1) from 1 toner container passing through 0.05 microns cartridge filter. Air hose and glass tubings were placed and the carboys were covered and then aerated. To eliminate chlorine in the prepared seawater, the trainee added 0.30g of Sodium thiosulfate (Appendix C, Plate 2). The presence of chlorine was monitored using orthotolodine (Appendix C, Plate 3). This was done by putting 2-3 drops, if the traces of the drops change its color to light brown there is still chlorine in the seawater.

Fertilizing

Before the inoculation of Nannochloropsis oculata, the seawater was enriched with nutrients using different types of fertilizer like F/2 medium, mQ medium, Conway medium, NA medium. The trainee used F/2 medium (Appendix C, Plate 4) as nutrient enrichment. The concentration of F/2 medium to seawater ratios was indicated in table 4 below.

Table 4. Concentrations of F/2 medium in seawater. Components Nitrate-Phosphate solution Trace metal-EDTA solution Vitamin solution Ratio 1 ml/L 1 ml/L 1 m/L

Inoculation In this activity, the trainee inoculated the starter culture with one liter of inocula, which was added to each carboy (Appendix C, Plate 5). The trainer informed the trainee that each culture has to be flashed with CO2 two times a day for five minutes.

Assessment of Algal Culture Two carboys of each batch of culture was checked and monitored for the presence of contaminants and algal density was also determined. Daily counting was done every day by the trainee. The equipment used was pipette, beaker, medicine dropper, thoma and microscope.

Small amount of sample was taken from the carboy using pipette and then transferred to a beaker. With the use of a medicine dropper, the trainee took sample of the culture, filled the thoma with it, and was then counted under the microscope. The thoma contained 16 blocks but 4 blocks was only counted. The formula used to get the density of the algae was A + B + C + D = ? / 4 x 16 x 104, where A, B, C, D is the number of algae in every block. Harvesting Harvesting was done during the exponential phase or early stationary phase which is on the 5th day.