FLORIA

FLORIA

Friday, 25 December 2015

Field Trip to Aquascape Paradise, Shah Alam

Mostly every subject related to Aquaculture courses has field trips to completed their syllabus  . On 29 October 2015, Aquatic Flora Culture went for a trip to Aquascape Paradise,Shah Alam. Estimated 30 KM from Kuala Lumpur.

The objective of this trip
  1.  To know type of aquatic plant that related to our subject
  2. To get more knowledge about aquatic plant local and imported from the shop.
  3. To get important information that can be apply in future uses.



Aquascape has two elements which are soft element and hard element. Soft element included plants and flowers. For the hard element consist of rock and wood (branches and roots)

The concepts that have been used in aquascaping such as:

  •         I.            Iwagumi Concept (many rocks used)
  •       II.            Biotope Concept (vegetation around pond used)
  •     III.            Dutch Concept (colorful and unique)
  •     IV.            Natural Concept (common concept)

Examples of aquascape designs.








We were given a tour around the shop. These are the pictures during the tour





This is a simple tutorial on how to create aquascape by CrystalRedeu











Thursday, 24 December 2015

Practical 4:Microalgae growth measurement

Introduction:
There are several methods that can be used to measure the microalgal growth. The examples of the mthod are estimation individual dry weight, optical density reading, and chlorophyl analysis. During this practical, we have done this 3 methods.

Material and method:
1. Estimation of individual dry weight:
    Dry wight of algal cells can be determined by filtering and drying algae from aliqquots of culture of known concentraation:


  • 3 duplicate counts was accurately determine their concentration of algal cutured to be sampled for dry weight analysis 
  • An exact volume [V] was filtered on pretared glass microfibre filters [W] using a Buchner set up connected to a vacuum pump (in triploicate). The filter was washed with a solution of ammonium formate (0.5 M) to remove salts.
  •   The filters were dried at 100◦C for 4h to volatilize the ammonium formate..It was then weighed on an analytical balance [W*-B*].
  •  The dry weight per algal cell was calculated according to the formula:DW (g/cell)=DWwc-DWbc/N × V 
  • The same procedure was followed with control filters on which an equal volume of 0.22 μm filtered seawater is filtered (in triplicate) [B].


2. Optical density reading:

     The optical density measurement was done with a spectrophotometer at 670nm. The spectrophotometer is fitted with a 10cm long cell. In this cell,we bring the sample to be measured. As a blank measurement we take the culture medium=seawater.

    When a bundle of light (I◦) is sent through an absorbing material,the bundle of light will have a lower intensity (I) when leaving the material. The absorption is dependent on the absorbing material in the solution (the algal cells) but also on the concentration and thickness of the layer (cell length).

This relation is described by the Lambert-Beer law : OD or A (absorption) = log10 I◦/I

Practically,we measured the absoprtion A of the seawater one time. We assume thisvalue to be I◦ since we are comparing it against the absorption of the algal cells in solution.

Then we set up a series of algal dilutions at 100%,80%,60%,40%,20%,0. The absorption of this series of dilutions is measured with the spectrophotometer and values are calculated according to Lambert-Beer law. The dilutions are also counted for number of algal cells with the haemacytometer counter chamber under the microscope.Using MS Excel we then put the values in a graph; Haemacytometer counts on the X-axis and OD values on the Y-axis. The a suggestion line is fitted and this can be used in the future to calculate cell concentrations for certain OD value.

3. Chlorophyl analysis:
  • 50ml sample was filtered through GF/F filtered.
  •    3 to 5 drops of MgCOᴈ was added to the sample as it is being filtered.
  • The edges of filter which are not coated with residue were trimed away.
  •  The filter was homogenized with 5ml acetone for 1 minute. 5ml of acetone was added more and grinded for 30 seconds.
  •   The sample extract was sampled in refrigerator in the dark for 1 hr.
  • The sample was centrifuged at 3000 rpm for 10 minutes.
  •  The absobance of sample extract was measured at 750nm,664nm,647nm, and 630nm.

Results

1)Estimation of Dry weight
Weight(g)
Filter with seawater
Filtre with microalgae
Before
0.12
0.11
After
0.13
0.13

2)Optical density analysis
%
Cell/ml
OD
0%
0
0
20%
9 000
0.1889
40%
150 000
0.3919
60%
260 000
0.5827
80%
360 000
0.7547
100%
1 000 000
0.9438


3) Chlorophyll analysis
Wavelength(wm)
630
647
664
750
Absorbance
0.0432
0.0567
0.0431
0.0204

Wednesday, 2 December 2015

Practical 5 : Propagation of seaweed in enrichment media

Introduction
Seaweeds are large and microscopic form. It is also marine macroalgae and has simple reproductive structures. Besides, seaweed do not have tissues for conduction (non- vescular). There are three basic parts of seaweed which are balde, stipe and holdfast. Then, there is various type of seaweed that played important role in environment system. The importance of seaweed is as bioremediator, food sources to human and animal, and biomolecules. Each group was given Glacilaria changii as sample for this experiment and different media each group.  



Sample
Glacilaria changii



Media
Sterile seawater, Von Stosch’s Enrichment (VSE) , Provasoli Enrichment Seawater (PES) media.



Bacteria culture
17 difference culture   *3 replicates



Steps
  1. Sample washed with tap water carefully.
  2. Sample was cut 1 cm by 1 cm
    Figure 1: cut sample 1 cm
  3. Then, weight sample one by one
    Figure 2: weight sample with electronic weighing 
  4.  wash sample with sterile seawater ( for 30 second)
    Figure 3: Sample was washed.
  5. Well –plate was filled with sterile seawater
  6. 104 CFU/ ml of selected bacteria was added (our group used bacteria 13,14,15, and 16
    Figure 4: Selected bacteria was added in each spaces. 
  7.  Next, seaweed was put in well- plate
    Figure 5: seaweed was put in well- plate.
  8. The sample was observed every seven days
Result  
               
              Day 1st 


                 Day 7th 
B1 sample buds already grow on it.


Buds grown
          Day 14th 
Almost all of the seaweed grow bud on it


One of the sample that buds grown on it


Another samples














Wednesday, 4 November 2015

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.





Monday, 2 November 2015

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.