Practical 1:PREPARATION OF CULTUR MEDIA

Practical 1

Objective:

1. To familiarize with the different microalgal sampling equipment
2. To know the procedure of preparing different algal medium culture

A) MICROALGAL SAMPLING EQUIPMENT:

• Water sampler:To collect water sample from water body such as river,lake,pond, sea. This equipment can be used to take water sample a different depth, based on purposes of the study. 
• Plankton net:To collect phytoplankton or zooplankton sample. Plankton net have different mesh size, which  can vary from 50 µm to 300 µm.
• Hydrolab:Multirpobes uses to measure the important parameter in water quality such as, pH, Dissolve oxygen (D.O), temperature, conductivity,salinity
• Refractometer: Equipment uses to measure the salinity of the water
• DO meter:Equipment uses to measure thedissolved oxygen content in the water
• pH meter:Equipment uses to measure the pH of the water
• Water depth sensor:Equipment uses to measure the depth of the water body
• GPS:Electronic devices uses to give a latitude and longitude of the sampling sites
• Sampling bottles:Uses to keep water sample
• Lugol solutions:Chemical uses to preserve the phytoplankton sample

B)PREPARATION OF MEDIA

There are many type of medium culture can be used as a media to culture microalgae. Each of the medium are specific to type of microalgae according to their classification.During this practical, only 5 medium was prepared in the laboratory. The medium are:

• Bold's Basal Medium (BB): Uses to culture freshwater microalgae

1. NaNO3, (10 g)
2. MgSO4·, , 7H2O,, (3 g)
3.  NaCl , , (1 g)
4. K2HPO4, , (3 g)
5. KH2PO,, (7 g)
6.  CaCl2·2H2O., (1 g)
7. Trace elements solutions that are required :
•  ZnSO4·7H2O, MnCl2·4H2O, MoO3, CuSO4·5H2O, Co(NO3)2·6H2O, EDTA, KOH, FeSO4·7H2O, H2SO4

• f/2 Medium: Uses to culture marine algae

1. NaNO3 75g 
2. NaH2PO4.2H2O 5.65g 
3. Trace elements (chelated) NA2 EDTA 4.16 g FeCl3.6H2O 3.15 g CuSO4.5H2O 0.01 g ZnSO4.7H2O 0.022 g CoCl2.6H2O 0.01 g MnCl2.4H2O 0.18 g Na2MoO4.2H2O 0.006 g 
4. Vitamin mix Cyanocobalamin (Vitamin B12) 0.0005 g Thiamine HCl (Vitamin B1) 0.1 g Biotin 0.0005 g 
5. Medium per litre NaNO3 1.0 ml NaH2PO4.2H2O 1.0 ml Trace elements stock solution (1) 1.0 ml Vitamin mix stock solution (2) 1.0 ml

• f/2 Medium + Silica (Guillard medium for diatoms)

1. NaNO3 75g 
2. NaH2PO4.2H2O 5.65g 
3. Trace elements (chelated) NA2 EDTA 4.16 g FeCl3.6H2O 3.15 g CuSO4.5H2O 0.01 g ZnSO4.7H2O 0.022 g CoCl2.6H2O 0.01 g MnCl2.4H2O 0.18 g Na2MoO4.2H2O 0.006 g 
4. Vitamin mix Cyanocobalamin (Vitamin B12) 0.0005 g Thiamine HCl (Vitamin B1) 0.1 g Biotin 0.0005 g 
5. Sodium metasilicate , Na2SiO3.9H2O
6. Medium per litre NaNO3 1.0 ml NaH2PO4.2H2O 1.0 ml Trace elements stock solution ,1.0 ml Vitamin mix stock solution ,1.0 ml

• Provasoli Enriched Seawater media (PES)

1. Solution I:
• deionized water, tris buffer, sodium hydroxide, glycerophosphate and thiamine (Vit. B1)
• Solution I:
2. Solution II:
• Fe act as EDTA complex; 1:1 molar.
3. Solution III:
• deionized water, disodium ethylenediamine tetraacetate, boric acid, ferric chloride, manganese sulfate monohydrate, cobaltous sulfate monohydrate and zinc sulfate, 7-hydrate
4. Solution IV:
• vitamins mixing with deionized water, vitamin B12, and biotin

• Von Stosch Enrichment (VSE)

• Solution I:
• deionized water and ammonium chloride
• Solution II:
• deionized water and sodium phosphate.
• Solution III:
• deionized water and Ferrous sulfate
• Solution IV:
• EDTA plus deionized water
• Solution V:
• manganese chloride and deionized water
• Solution VI:
• mix of vitamin thiamine, biotin B12 and deionized water.