Optimization Simplex Method Green Algae Botryococcus Braunii Biology Essay

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The basic simplex method are applied for different type of culture media composition for autotrophic Botryococcus Braunii .it was used to increase on the kinetic growth and cytogenesis to effect nutrient concentration and algae growth in different nutrient medium concentration. During geographically study to increase salinity ratio in phage growth and lag phage are also depend on the nutrient. The simplex method is used to solve problem in a linear programming. Botryococcus Braunii is use to growth bold base medium with 3 fold nitrogen and vitamin, modified (3N_BBM+V).algae Botryococcus Braunii are growth in bold based medium and three fold nitrogen.

Algae growth is used same medium and also as well to use in different nutrient concentration. Botryococcus Braunii has greatly effect to the nutrient and also affects the 680nm wave length. Algae effect on different nutrient concentration and effect on growth of algae. Sodium chloride, trace element solution and calcium chloride are less effect on algae growth medium. Botryococcus Braunii has growth in standard medium bold based medium and three fold nitrogen. 3N-BBM+V medium are used only fresh green algae. This medium has specific nutrient concentration if we changed nutrient concentration algae does not grow on different medium. Botryococcus Braunii are used in pharmaceutical industry and as well used in textile industry. Spectrophotometer was a initial measurement on algae growth medium and take a photograph through electronic microscope. Linear simplex method optimization was used to solve problem during the practical.

INTRODUCTION:

Algae are the term used for some lower plants and often unrelated groups of microorganism that are able to perform photosynthesis. Photosynthesis is performed in parts of the cell called chloroplast. They have different shapes and colors and found in different organism. Not at all these organisms are green .diatoms Chlorophytes and Dinoflagellates have yellow to brown chloroplast. They are brown algae (phaeophyta), Red algae (Rhodophyta) and many other groups of unicellular green algae (Scendesmus Dimorphus and Botryococcus Braunii). Very diverse groups of freshwater algae are the Chlorophytes (green algae) based on the compounds of the photosynthesis pigment and several other characteristics they seem closest related to plants.

Botryococcus Braunii

Scientific classification

Kingdom:

Plantae

Division:

Chlorophyta

Class:

Chlorophyceae

Order:

Chlorococcales

Family:

Dictyosphaeriaceae

Genus:

Botryococcus

Species:

B. Braunii

Binomial name

Botryococcus Braunii

Kützing

WEB diagram 1:

Green color from chlorophyll a and be in the same properties as a higher plant beta-carotene and xanthophylls .some green algae (Botryococcus Braunii) to be progenitors of the higher green plant. The colonies and cell of Botryococcus Braunii laboratory culture were examined and bloom cell isolated from lake. The cup like green algae cell represent a parental cell were located on the upper surface green belt. The arrangement of green cell in laboratory culture was consisted group of 2-4 cell formed unite and these unit joined in cluster. Biomass photosynthesis is a very main source of energy. It is converted and reduces carbon dioxide from the environment .species Botryococcus Braunii is produced and accumulates hydrocarbon at 30-40% on dry weight. The growth of Botryococcus Braunii is sewerage secondary treatment in batch system .STS have represented to be an effective growth rate for microalgae and inorganic compound in STS were examined in batch culture .Botryococcus Braunii have used in domestic sewerage for cleaning water and it is also main source of energy in future. It is also used for produce fuel. They are normally associated harmful algae and public health found that in the aquatic environment in the world wide. Dinophytes, chlorophytes and cynobacteria are found in aquatic ecosystem in bloom forming algae. These are produced saxitoxin, microcrystal and some active compound produced with allelepathic activity. Botryococcus Braunii green alga colonel is usually hydrocarbon content and renewable hydrocarbon Botryococcus Braunii algae. This hydrocarbon has used as an alternative bio fuel and diesel and Botryococcus Braunii massive growth rate can give rise to obvious bloom in aquatic ecosystem in the world wide. Botryococcus Braunii species and it is a new variety present name showa. This isolated from 1980 from illy culturing tank in the California green house. Illy pond culture was collected orange colour colony from pond and without washing colonies were diluted in enriched hydrocarbon medium culture orange colonies were selected by means of a 40 lower power magnification in under electronic microscope.

Optimization basic simplex method is used to apply for different type of media composition culture of Botryococcus Braunii. The simplex method is used to problem solving in a linear programming. This method was discovered by George in 1947 to use adjacent test vertical of the possible test in sequence and objective improves. Green colonel micro alga Botryococcus Braunii is unusually rich alternative sources of the hydrocarbon and other chemical. It can be consisted up to 75% of Botryococcus Braunii dry mass. The review detail various form of Botryococcus Braunii and include physiology or microbiology, hydrocarbon production and other compound by culture of algae method, downstream recovery and processing of algae hydrocarbon. Botryococcus is found normally green colors in the world. it consist of the high hydrocarbon and used to waste water treatment have attracted increasing attention in the recent year .

Colonel algae ample Botryococcus Braunii collected from various locatable were grown in laboratory and consist for their hydrocarbon contents a morphology form a few differences appeared between the ultra structures of the sample nature of their hydrocarbon. it remain and unchanged at any stage of growth to allow the distinction.

Different from the water solvent was tested for biocomtability and hydrocarbon recovery under certain condition with Botryococcus Braunii rich hydrocarbon. It can be used to secondary treatment of waste water treatment in a continuous bioreactor treatment. the hydrocarbon content of algae grow on STS compare well that of algae STS would be appeared to surface as an appreciating medium for continuously and remain able growth. Now days a wide range used to compare such as polyphone, polyunsaturated, fatty acid obtain from wine.

Method basic simplex optimization:

Method basic simplex can be understands and easily applies. Initial trial is beginning optimization. They are spread out efficiency and number of trial initial is equal to a plus value one. These trials are called first simplex optimization. It has basically mathematically approach and optimum condition e.g. triangle, line and tetrahedral.

http://www.grabitech.se/simplex_b.htm

http://www.grabitech.se/simplex_b.htm

A new variable control is calculated by reflection into the control space variable opposite undesirable results. The new trial has to need change one favorable trial in the simplex. This lead is to be a new trial. Simplex is normally moved to more favourable condition. This problem is nicely avoided by choosing second favorable and moving away from it. Multisimplex algorithm calculated is derived into chart. It explains with different level and label as well.

W= favorable trial

B = unfavorable trial

NW= second less favorable trial

WEB DIAGRAM 2: http://www.grabitech.se/simplex_b.htm

Material and method:

Culture flasks and shaker:

Klyuyers and perquin was first type used culture in flask and shaker in 1973 and now it has been used for the microbial culture ,animal and plant cell flask culture can be used in large series in order to appraise the productivity of microorganism ,composition of media and raw material or control the quality of inoculums.

Culture flasks:

"Size and shape laboratory culture on shakes are carried out using test tubes, flasks and bottle of different types and size. The most widely used being Erlenmeyer and conical flask of 500m in volume"

"Methods in industrial microbiology B, SIKTA"

Some microorganism culture especially bacteria is done in test tube using less volume of medium. It can be used for determination of strains, identification of microorganism contamination. Algae culture is in flask using large volume of medium and it can be used by algae culture growth, large volume growth and contamination of algae. It is made of glass. There are advantage, observing growth rate easy cleaning, sterization and low prize. The glass used must not release trace element even when not alkaline, heat and acid. This instrument can be washed in automatic washing device and it has a labour cost of washing in big lab research project. Now a day in laboratory scale glass flask have changed to polypropylene plastic. Plastic flask has cheaper and discarded after single use. The plastic are completely inert inhabit culture growth.

Flask closure:

During practical flask closure has a little attention. It is important to the aseptic aerobic culture. The flask closure effect to the aeration condition in the culture and cotton wool plugs are used to close flask as a represent from aeration as oxygen must first diffuse through before reaching the surface of the liquid in the flask.

Shakers:

"The principle feature required of shakers are quiet reliable and fault free run without vibration. Even during long term operation, possibility of charging the frequency and amplitude of exertic movement in rotator shaker simple and speedy replacement of shaker trays and low operation expensive"

"Methods in industrial microbiology by B.SIRYTA"

ROTATORY SHAKER:

The rotatory movement shaker is derived from electrical motors via v-belt. It is attached the shaker frame .it can be used to large number of oxygen transfer rate and it have usually frequency 7HZ in 15minut.

Culture procedure:

Culture flask are placed tray with various type of different shape and size flask. The vertically position of flask have higher rate of oxygen transfer. In laboratory scale is used up to four scale tray for saving time in working space.

All nutritional are required in a medium .media will provide and good support to the growth of microorganism. This chapter was explained technique which enables the growth of selective algae B.Braunii and avoids from risk contamination unwanted species.

IMPORTANT 3N-BBM+V ALGAE CULTURE:

3N-BBM+V have two possible methods to describe to use sol gel technology. It used to immobilize microorganism living with thin silica layer and structure preparation of both biocer variant. Algae and bacteria have to use silica for storage stability and growth. Biological oxide matrix system to combining ceramic layer and sol gel convenient procedure are used generally in high concentration of alcohol in living cell. This method is used in textile industry to growth chlorella vulgure. This species have efficient method to remove color from textile. This species have growth within 15 days in same media culture and biological function to help remove colour in textile. It has naturally procedure to use for many years in textile industry.

Batch culture and continuous culture have certain changed environmental condition, nutrients

, concentration, product and specific growth. Continuous and batch culture have expensive in laboratory. It's used only industrial level because they are high quantity of production. 3N-BBM+V have used in laboratory scale to algae growth. it has depend on the flask shake method and it has very cheap and easy procedure for algae growth. two symbolize method for quantities estimation of algae soil, method dilution culture and chlorophyll extraction were compared using soil sample collected from irrigation land in the flood plain culture dilution method was almost same. The number of colonies logarithm and dilution method was almost same. This method used only seasonal. This used algae growth and extraction of lipid and after that lipid is converted to the oil and energy sources. It has big energy sources in future.

3N-BBM+V CULTURE MEDIUM BOTRYOCOCCUS BRUNII:

HEALTH AND SAFETY:

Please read properly health and safety procedure libratory instruction and print out coscch laboratory work. Lab assistant and professor must sign coscch work otherwise you cannot start project.

APPRATUS and chemical:

25 100ml conical flask, 10 250ml conical flask, algae medium 3N-BBM+V, trace element, sodium nitrate, calcium chloride , magnesium sulphate, potassium sulphate, potassium sulphate hydrate, sodium chloride, ferric chloride, manganese chloride, copper chloride,vitamine B1 vitamineB12 and algae culture Botryococcus Braunii

Spectrophotometer, burette, 1000um pipette, 10 1litre bottle, weight machine, cotton, silver paper and shaker machine.

"All essential nutrition is required in algae medium and medium will provide to food support during the algae growth. Botryococcus Braunii algae growth in bold basal medium with three fold nitrogen and modified vitamins medium used only green algae". Made an algae growth medium in some following steps:

Made a trace element solution "Add to 100ml of distilled water o.75 g Na2EDTA and the

Mineral inn exactly as given below in table"

salt

Quantity in mg

 FeCl3.6H2O

97

MnCl2.4H2O

41

 ZnCl2.6H2O

5

 CoCl2.6H2O

2

 Na2Mo4.2H2O

4

Vitamin B1:

Put 0.12 g thiaminhydrochloride in distilled water 100 ml and sterile filtered as well

Vitamin B12:

Cyanobalamin 0.1 g in distilled water 100 ml, took 1 ml of the solution and put 99 ml distilled water . Sterile filtrate as well

Made algae medium in different nutrient concentration. Table as given below:

Stock solution in g/1000 ml water

For 1 litre medium quantity in ml

NaNO3

30

MgSO4.7H2O

10

NaCl 

10

 K2HPO4.3H2O

10

 KH2PO4

10

CaCl2.2H2O

10

Trace element solution

6

VitamineB1

1

Vitamin B12

1

Filled up 1 litre conical flask in distilled water.

Autoclave in one day in normal fridge temperature.

Next day put sterile I ml vitamin B1 and vitamin B12 in media solution.

Prepared 7 small conical flasks filled up to 100ml solution.

Cultivated Botryococcus Braunii algae in lake.

Put I ml algae in 2 small medium bottle and kept in rotatory shaker.

Took a absorbance reading with help spectrophotometer and carry on next few days to observe algae growth.

Prepared two smaller conical flask medium with algae culture and followed same last experiment.

Checked all four sample proper algae growth and selected one sample to follow next simplex optimization based practical.

Diagram during practical section as given below:

ISU's159 ISU's164

12-algae sample four to use for algae growth in different nutrient concentration. Algae sample put 1 ml one by one every sample and take an absorbance reading in spectrophotometer at 680 n m.

13-prepared a different nutrient concentration in normal medium. First three sample with extra 1ml sodium nitrate, sodium chloride and magnesium sulphate. Followed same procedure.

14-The spectrophotometer have to be there set up and prepared to use. The act or process of exciting wavelength is 680nm. All work will be much more meaning realizing algae invisible in naked eye. Microscope was used to observe and take it some picture. Took some algae sample on glass slide and covers with cover slip.obsevered in under microscope. Microscope fixed with camera into computer and took a snap.

Took all sample go to biochemistry lab and I got it some snap 10 mega pixel camera in electronic microscope.

15- made a three more different nutrient concentration e,g potassium phosphate, potassium dehydrate and trace element. Took all of them 1ml in normal medium solution.

16- Prepared a calibration graph in different nutrient concentration in 3N-BBM+V .the table as given below.

Stock solution in g/1000 ml water

For 1 litre medium quantity in ml

NaNO3

30

MgSO4.7H2O

10

NaCl 

10

 K2HPO4.3H2O

5

 KH2PO4

5

CaCl2.2H2O

10

Trace element solution

6

VitamineB1

1

Vitamin B12

1

17- Poured one ml algae sample in calibration bold based medium three fold nitrogen based and calibration reading with help spectrophotometer at 680nm.

18- Made a two more algae culture sample with extra vitamin B1 and extra vitamin B12 in ml in normal medium solution.

19- All the remaining sample took picture in electronic microscope.

Result:

Botryococcus algae growth in normal medium and different nutrient concentration thus I was divided in two experiments. Experiment 1 was explain algae growth in normal standard medium and experiment no 2 was explain algae growth in different nutrient concentration.

Experiment no 1: ALGAE GROWTH IN NORMAL MEDIUN N-BBM+V:

First experiment was showed that algae growth in normal basal based medium with three fold nitrogen. We have already explained how to make algae medium. This medium was used for green algae because algae Botryococcus Braunii sample was used in different experiment to examine algae effect in normal and different nutrient concentration. Spectrophotometer was used to examine algae growth. BOTRYOCOCCUS BRUNII algae are growth in bold based medium three fold nitrogen. Algae growth is also effect on the absorbance at 680nm. In the present study algae Botryococcus Braunii growth, photochemical process and under photosynthesis process were higher compared to the under nutrient concentration. Algae Botryococcus Braunii has better growth in normal medium culture.

Calibration curve algae growth against days

DAY

SAMPLE1680nm

SAMPLE2 680 nm

SAMPLE3 680nm

SAMPLE4 680nm

1

0.158

-

-

-

2

0.222

0.268

-

-

3

0.096

0.151

-

-

4

0.428

0.223

-

-

5 AGAIN START

0.452

0.248

0.00

0.00

7

0.713

0.307

0.111

0.296

8

0.545

0.368

0.568

0.365

9

0.446

0.683

0.419

0.112

11

0.613

0.954

0.494

0.462

13

1.114

0.650

0.648

0.751

14

1.078

1.020

0.955

0.741

15

1.134

1.085

1.088

0.887

17

1.191

1.238

1.093

0.967

19

1.486

1.531

1.203

1.190

21

-

-

-

1.237

22

-

-

-

1.322

24

-

-

-

1.127

First graph was shown algae medium did not proper growth in normal medium.Botryococcus Braunii was effect in wrong medium may be was something wrong during practical. Thus experiment was started again algae growth in normal medium. We prepared two more sample was used to following practical.

Picture algae Botryococcus Braunii sample under electroni microscope 100magnification in given below.

normal%20last

Experiment 2: algae Botryococcus Braunii growth in different nutrient concentration and algae medium:

Experiment no2 was important part for this practical. Optimization simplex method was used growth in algae Botryococcus Braunii. Botryococcus Braunii was growth effect in different nutrient concentration. Nutrient concentration was greater effect to algae growth. Algae was growth in normal medium but algae growth medium changed and as well algae growth rate. We explained algae growth in different nutrient concentration.

Role sodium nitrate and sodium chloride in algae (Botryococcus Braunii) growth medium:

Sodium nitrate and sodium chloride was important role in algae growth medium. We used algae growth in three different medium for example normal standard medium and sodium nitrate and sodium chloride 1 ml extra in normal medium. Salinity was important role in algae growth. Spectrophotometer was used to measure algae medium.

TABLE 2: calibration curve ALGAE GROWTH DIFFERENT NUTRIENT CONCENTRATION( SODIUM CHLORIDE AND SODIUM NITRATE) ABSORBANCE AGAINST DATE

DATE

SAMPLE A NOADDITION 680nm

SAMPLE B SODIUM NITRATE 680nm

SAMPLE C SODIUM CHLORIDE 680nm

17/11/09

0.233

0.180

0.161

19/11/09

0.425

0.258

0.244

20/11/09

0.519

0.281

0.318

24/11/09

0.788

0.428

0.455

3/12/09

1.668

0.996

1.198

Role trace element solution and potassium phosphate in Botryococcus Braunii algae medium:

Botryococcus Braunii algae were growth in different nutrient concentration. Some nutrient concentration was less and some time more effect to algae growth. Growth Botryococcus Braunii was used another experiment for different nutrient concentration e.g. sample d no addition, sample e KH2PO4 and sample f trace element solution. Algae growth was grown in normal medium 3N-BBM+V comparison another nutrient concentration. If you changed normal medium composition and algae growth are also effect. Spectrophotometer was a first instrument to investigate algae growth absorbance. If you made slide algae sample and checked electronic microscope as well take picture. We examine algae growth is also effected on this sample. Sample A was more growth and good absorbance another sample. Trace element solution extra in normal medium culture was also affected on the growth biomass but potassium phosphate was not proper growth and less algae growth comparison another sample.

TABLE 3 : CALIBRATION CURVE ALGAE Botryococcus Braunii growth in different nutrient concentration trace element and potassium phosphate against days:

DAYS

SAMPLE D (NO ADDITION) 680 nm

SAMPLE E (KH2PO4) 680 nm

SAMPLE F (TRACE ELEMENT) 680 nm

0

0.00

0.00

0.00

4

0.205

0.166

0.241

6

0.380

0.256

0.409

10

0.799

0.593

0.875

Role new medium and trace element solution to growth in algae Botryococcus Braunii :

Trace element solution did not greater effect to algae growth if we changed algae medium composition and also effect to growth. Sometime algae did not growth in different medium. Algae always growth in specific medium.

Calibration curve Botryococcus Braunii algae growth against different nutrient concentration:

DAYS

TRACE ELEMENT SOLUTION ABSORBANCE AT 680nm

Normal medium absorption at 680 nm

New medium absorption at 680nm

0

0

0

0

1

0.034

0.101

0.057

4

0.267

0.277

0.075

6

0.342

0.507

0.116

8

0.395

0.603

0.141

Role calcium chloride in algae growth Botryococcus Braunii medium:

Calcium chloride was less effect to algae growth. Actually calcium chloride had important part for algae growth medium composition and it is used for algae growth medium. Spectrophotometer was used to measurement for algae growth.

DAYS

I EXTRA CALCIUM CHLORIDE 1ml 680nm

NORMAL 680nm

1

0.173

0.193

4

0.318

0.354

5

0.437

0.573

Role vitamin B1 and vitamineB12 to effect in algae (Botryococcus Braunii) medium :

Vitamin B1 and vitamin B12 are important part for algae growth medium composition and these are also important part for algae cell composition. Vitamin B1 AND B12 was effect on algae growth medium but it has less effect to algae growth medium.

DAYS

K VITAMINE B1 680nm

L VITAMINE B12 680nm

1

0.203

0.116

4

0.308

0.504

6

0.408

0.706

Discussion:

BOTRYOCOCCUS BRUNII algae are growth in bold based medium three fold nitrogen. Algae growth is also effect on the absorbance at 680nm. In the present study algae botryococcus brunii growth, photochemical process and under photosynthesis process were higher compared to the under

nutrient concentration. Algae bortyococcus brunii have better growth in normal mediun culture. If you change nutrient concentration in normal medium and definitely change to the algae growth .in the general nutrient concentration profile changes with the physiological state of the culture. We used spectrophotometer 680nm wavelength to take absorbance reading. This reading was telling you algae growth or not .it means to say this experiment depend on the photosynthesis process.photsynthesis is one of the most important biochemical procession the plant or green algae. In the process of photosynthesis green algae absorb carbon dioxide from the atmosphere and make food. The occurrence of photosynthesis process by the green colour.the spectral distribution of the response function for photosynthesis might give impression that visible radiations in the green range central 680nm contribute very little to the photosynthesis process and therefore is of minor important. Just contrary has been demonstrated by experiment. It is precisely this green radiation that yields the greater productivity and efficiency in densely populated arrangement for investigation into the yield on algae. First of all I did algae (botryococcus brunii) growth in specific nutrient concentration .i took absorbance reading after every two days. I got problem during start practical because I was doing use wrong spectrophotometer actually I put wrong way cuvett in spectrophotometer.lab assistant change my spectrophotometer. I sort out this problem after that I got a good result was carry on my project practical based on these basic simplex method. There Are these type of method e, g linear, triangular and tetrahedral. I used 680nm shorter wavelength depend on the photosynthesis process. Algae growth was in specific medium 3N-BBM+V with specific nutrient medium concentration. I followed these practical with different nutrient concentrtion.botryococcus brunii growth and production of its constituent's hydrocarbon, fatty acid were influenced by different level of salinity. The growth of hydrocarbon rich botryococcus brunii was studied under certain condition using batch culture and continuous culture. Large difference in the physiological change of botryococcus brunii was achieved in batch culture with various dilution rates. The possible effects of these variations upon hydrocarbon and also on the production of excellular polysacchrite were examined. The relationship between the physiological state of botryococcus brunii and its hydrocarbon or polysaccrite production were discussed and compare with those generally derived in unicellular algae. Algae botryococcus brunii sample 4 was showing more growth comparison another three methods and spectrophotometer to use for confirm algae growth. Sample no 4 was used to another experiment.

Algae have effect on the different nutrient concentration. Some halophytic algae will be growth in high salt concentration and it is also adapt in high salt concentration. There are many growth factors to effect on algae growth and as well effect on the environmental condition e.g. PH, temperature and salinity. Growth Botryococcus braunni was used on another experiment. Algae growth in three different medium. I was labelled A no addition, B with sodium nitrate 1ml extra in normal medium and C with extra 1ml extra sodium chloride. Three bottles was kept in fridge for 2 days after that I observed algae growth on spectrophotometer at 680nm. I got a result algae growth effect on nutrient concentration. I carried on these procedures for one week after that I took picture on electronic microscope 100 magnifications. Algae were more growth in addition sample because botryococcus braunii was not proper growth in different nutrient concentration. Salinity has also effect in different species of algae. Sodium nitrate has more effect on the algae growth. Sample b have less algae growth comparison to sample c .sodium chloride is gradually growth to compare sample. Botryococcus Braunii was growth and production carotene, fatty acid and hydrocarbon partial by different level of salinity. Botryococcus Braunii culture was obtained in unicellular algal clone in the fresh water pond. Algae pigment and other aspects were studied in the chemical composition. if we change chemical composition culture and it also effect algae growth .green algae was a relative studies of pure and mixed culture and their decompose residue was performed by using extra nutrient .Spectrophotometer was performed by using primary analytical method.Growth botryococcus brunii was used another experiment for different nutrient concentration e.g. sample d no addition, sample e KH2PO4 and sample f trace element solution. Algae growth was grown in normal medium 3N-BBM+V comparison another nutrient concentration. If you changed normal medium composition and algae growth are also effect. Spectrophototer was a first instrument to analyse growth absorbance. If you made slide algae sample and checked electronic microscope as well take picture. We examine algae growth is also effected on this sample. Sample a was moiré growth and good absorbance another sample. Trace element solution extra in normal medium culture was also affected on the growth biomass but potassium phosphate was not proper growth and less algae growth comparison another sample.Sample d no addition was not effect on algae growth and sample f trace element solution algae growth was normally less then normal medium. Sample e new medium. Algae culture made it again but potassium phosphate quantity less comparison to standard value and algae was not grown this medium. We examined in under electron microscope and spectrophotometer at 680nm. Sample e was not shown to algae grow comparison then another two sample d and f.The effects of culture exudates obtain botryococcus braunii green algae on cell suspension and biomass production was examined. If algae culture changed as well as biomass algae growth change as well. Fresh green algae are growth in standard solution e.g. 3N-BBM+V basal based medium only growth for botryococcus green algae and another green algae. It has specific nutrient concentration if we changed nutrient concentration on normal medium. Algae growth was also effect on algae biomass. Spectrophotometer and electronic microscope was examining algae growth. Calcium chloride is a main source to help for algae growth but calcium chloride did not effect on algae growth. Botryococcus braunii was growth in normal medium as well one sample I ml extra calcium chloride in normal medium to use for algae growth. Algae botryococcus braunii growth did not effect on calcium chloride. Spectrophotometer was used to confirm algae growth in normal an as well extra calcium chloride. Electronic microscope was used to take picture liquid algae sample. This picture proved that algae did not effect on calcium chloride but necessary in standard fresh algae. Medium.Algae Botryococcus braunii are normally found in natural environment e.g. fresh water lakes. It is naturally colour dark green and algae growth in some certain condition. If we used algae growth in laboratory. Bold based medium 3 fold nitrogen source have a standard culture method to use algae growth in laboratory. E.g. made two sample 1 vitamins B1 and sample two vitamins B12 I extra I ml in normal medium. Vitamine B12 was greater effect on algae growth comparison then vitamins B1 algae sample. Spectroscopy and electron microscope was an initial confirm to algae growth in culture medium. Vitamine was used in normal medium to use algae growth as well as in pharmaceutical industry.

Future work:

Acknowledgement

First of all I would like to thanks to almighty the most compassionate and most merciful.

I would also like to thank to my project's supervisor NEIL MORGAN for his support, encouragements, motivations and guidance throughout the project.

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