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.

Marine Macrophytes at Teluk Pelanduk, Port Dickson, Negeri Sembilan

Teluk Pelanduk, Port Dickson

Seagrass

Seagrass are actually not ‘grasses’ at all, as they do flower. Like land plants, seagrass produce oxygen. The depth at which seagrass are found is limited by water clarity, which determines the amount of light reaching the plant. Light is required for the plants to make food through photosynthesis.

Teluk Pelanduk ecosystem growth some species of seagrass which can be seen in lower tide only because of low water turbidity.

Samples of seagrass that we found during the trip.

1. Halodule pinifolia

Halodule also known as needle be short to very long. The leaves have three parallel veins which can be quite distinct. In most, the central mid-rib vein is quite prominent. The leaves emerge from thin rhizomes (underground stems) which have fine rootsseagrass is a dominant species in Teluk Pelanduk. Their habitat usually on the edges of the sand bars towards the low water mark.

2. Enhalus acoroides

Enhalus acoroides has the longest leaves of seagrasses found on our shores. The strap-like leaves are 1-2cm wide and 30cm-1.5m long. The edges of the leaves are slightly rolled. The leaves have air channels in them.

This seagrass has thick rhizomes (underground stems) that are densely covered with the stiff black fibrous strands, which are the remains of old leaves. The rhizomes have also many cord-like, hairless roots. The roots also have wide air-channels.

3. Thallasia hemprichii

The seagrass has strap or curved, sickle-shaped leaves (0.5-1cm wide and 7-40cm long, usually less than 25cm). The tips are usually rounded and smooth. The leaves may appear speckled due to tannin cells that appear red, purple or dark brown. It has thick rhizomes (underground stems) about 2-4mm in diameter which are white or pink. The rhizomes have air channels and usually have obvious node scars that are triangular with persistent leaf sheaths. Shoots emerge from these rhizomes, each shoot with 2-6 leaves encased in sheaths about 3-8cm long.

4. Halophila ovalis

The seagrass has oval, spoon-shaped leaves and is sometimes also called 'paddleweed' or fan seagrass. It comes in a wide range of sizes (0.5-1.5cm wide and 0.5-2.5cm long) and shapes from oval, to nearly oblong or spoon-shaped. The leaf edge is smooth with no serrations, there is a vein just within the leaf margin (intramarginal vein). The leaf has obvious cross veins (4-25) and is held on a long thin stalk. It has thin, smooth, white rhizomes (underground stems) about 2mm in diameter. The leaves emerge in pairs from these rhizomes. The emerging shoot is encased in a pair of transparent scales.

Seaweed

"Seaweed" is the common name for countless species of marine plants and algae that grow in the ocean as well as in rivers, lakes, and other water bodies.

Some seaweeds are microscopic, such as the phytoplankton that live suspended in the water column and provide the base for most marine food chains. Some are enormous, like the giant kelp that grow in abundant “forests” and tower like underwater redwoods from their roots at the bottom of the sea. Most are medium-sized, come in colors of red, green, brown, and black, and randomly wash up on beaches and shorelines just about everywhere.

The vernacular “seaweed” is a bona-fide misnomer, because a weed is a plant that spreads so profusely it can harm the habitat where it takes hold. (Consider kudzu, the infamous “mile-a-minute vine” that chokes waterways throughout the U.S. Southeast). Not only are the fixed and free-floating “weeds” of the sea utterly essential to innumerable marine creatures, both as food and as habitat, they also provide many benefits to land-dwellers, notably those of the human variety.

Samples of seaweed that we found during the trip.

1. Glacilaria sp.

Gracilaria is a genus of red algae (Rhodophyta) notable for its economic importance as an agarophyte, as well as its use as a food for humans and various species of shellfish.