FORM 2
THE PATENTS ACT, 1970
(39 of 1970)
&
The Patents Rules, 2003
COMPLETE SPECIFICATION
(See section 10 and rule 13)
1. TITLE OF THE INVENTION: "CONTAMINATION FREE CULTiVATION OF ALGAE"
2. APPLICANT(S)
(a) NAME: ABELLON CLEANENERGY LIMITED
(b) NATIONALITY: Indian
(c) ADDRESS: 10th Floor, Sangeeta Complex, Near Parimal Crossing, Ellisbridge, Ahmedabad- 380 006, GUJARAT, INDIA, Telephone No.: +91-79- 26563331, Fax No. +91-79-26408053,
E-mail; abellon(5)abelloncleanenergy,com
3. PREAMBLE TO THE DESCRITION
COMPLETE
The following specification particularly describes the invention and the manner in which it is to be performed
4. DESCRIPTION (Description shall start from next page) AS PER ATTACHED SHEET
5. CLAIMS AS PER ATTACHED SHEET
6. DATE AND SIGNATURE (to be given on the last page of specification)
7. ABSTRACT OF THE INVENTION (to be given along with complete specification on the separate page)
AS PER ATTACHED SHEET
Note:
*Repeat boxes in case of more than one entry
*To be signed by the applicant(s)or the authorized registered patent agent
*Name of the applicant should be given in full, family name in the beginning
*Complete address of the applicant should be given stating with postal index no. / code, state and country
'Strike out the column which is/are not applicable

FORM - 2
THE PATENTS ACT, 1970 (39 OF 1970)
COMPLETE SPECIFICATION (SEC-10, RULE 13)
TITLE: "CONTAMINATION FREE CULTIVATION OF ALGAE"
Applicant: ABELLON CLEANENERGY LIMITED
10th Floor, Sangeeta Complex, "Nit. Parimal Crossing, Ellisbridge, Ahmedabad-380 006, Gujarat State. India,
The following specification particularly describes the invention and the manner in which it is to be performed.

FIELD OF INVENTION:
The present invention relates to Cultivation of contamination free marine micro algae irrespective of cultivation system (flask, open pond, bioreactor etc). More particularly the present invention pertains to application of quinine preparation to micro algae cultivation medium at various concentration and maintain contamination (grazer-particular ciliate) free cultivation.
BACKGROUND OF THE INVENTION:
Algae essentially harness energy via photosynthesis. They capture CO2 and transform it into organic biomass which can be converted to energy. Micro-algae are unique photosynthetic organisms having the capability to accumulate lipids, proteins, and carbohydrates in substantial quantities and thrive in high salinity water relying on C02, sunlight and micro-nutrients for its growth. The use of algae in biotechnological research and industry is significant. Algae play roles as biocatalysts for the production of food, chemicals and fuels and they are becoming important in the development of solar energy technology, biodegradation and bioremediation. In addition, some species of algae are eaten directly by humans.
There has been a constant need for different and efficient sources of renewable energy forms. We live in an area with a great deal of biomass, which could be used to produce renewable energy. The continuous use of fossil fuels by mankind has caused immense damage to environment. The drastic increase in the emission of CO2 caused by burning fossil fuels has been identified as the major reason for the change of temperature in the atmosphere causing various global climate events such as global warming. Looking into a broader aspect of utilizing the biological forms for meeting the energy crunch and also its wide range of potential applications Algae has served as one of the most potential form of biological feedstock.
Algae is because of its desirable properties has been preferred due to 1) comparative higher growth rates, 2) permit the use of non-arable land and non-potable water,3) use far less water, 4) does not displace food crops cultures .5) production is

not seasonal, 6) can be harvested daiiy and 7) ability to use nutrient from waste stream.
The different biomolecule composition of microalgae broadens its range of applications thereby giving it a niche over all other biomass. Micro algae plays vital role as bio-catalyst for the production of food, carotenoids .dyes, fine chemicals, health food , biofuels and therapeutic supplements such as β-carotene, astaxanthin, polyunsaturated fatty acid etc.
Micro algae convert solar energy into chemical energy through photosynthesis and store this energy in the form of lipids, carbohydrates, and proteins. These cellular constituents can be used to derive various value chain applications.
Micro algae can be cultivation in open type cultivation as well as close type (photobioreactor) condition. In close type cultivation high cell yields can be achieved as we can control all growth parameter. While in case of open raceway pond chances of spoilage of culture, as in open pond growth factor are not in controlled manner. But both of the above mentioned methods have one common problem is contamination of grazer(in particular ciliate). Due to such contamination, growth/yield of micro algae is reduce per unit. And thus commercial production of micro algae is not economic. Hence, there is a need to come up with some solution to inhibit the growth of the said grazer without harming the micro algae growth & their biomolecular content.
PRIOR ART
In US20090298159Al, (Publiced on 2009-12-03) a method is provided to produce biodiesel from algae using a two-stage, autotrophic and heterotrophic cultivations of chlorella for biodiesel production. This method includes a sequence of procedures: cultivating photoautotrophic algae, concentrating cells and then transferring them to a fermentor for heterotrophic cultivation. During the photoautotrophic cultivation stage, the culture is exposed to a light source, such as sunlight with carbon dioxide obtained from a carbon dioxide source or from air. antibacterial agents may be added to prevent contamination from undesired microorganisms. Organic carbons are added during heterotrophic cultivation stage. Fermentation conditions are optimized for

maximizing lipid synthesis. High biomass is achieved to about 108 g/L with lipid content reaching about 52% of dry cell weight. After cultivation, biodiesel is made through extraction and transesterification of algae lipids.
OBJECT OF THE INVENTION
The main object of the present invention is to inhibit the growth of grazer(in particular ciliate) which tackles most of the issues, currently associated with algae cell growth in both closed and an open algae cultivation systems.
Another, object of an invention is to produce higher micro algal biomass in order to achieve higher biomolecule content of lipid carbohydrate or protein.
Yet another, object of the invention is to produce continuous production of algae (In particular micro algae) biomass for production of bio-fuel and bio-products that can be obtained in sufficient quantities throughout the year.
Further, object of an invention is to raise the over all process cost efficiency.
DETAIL DESCRIPTION OF INVENTION
In accordance with the present invention, a method is provided to contamination (in particular ciliate) free micro algae cultivation. Further embodiment of the invention are directed to cultivate micro algae free from other protozoa and bacteria which inhibit the growth of micro algae in open as wei! as close cultivation system.
In, the first stage algae (in particular micro algae ) is cultivated either in a closed system or in an open system. Grazers (in particular ciliate) is observed to be inhibiting the growth of algae cells cultivated in any system. The said grazer inhibits the growth of algae by reducing cell density of algae through engulfing algae cells grown in cultivation system. By, adding quinine preparation at this stage lowers down the activity of the said grazer and inhibits the growth of the said grazer.
The, embodiment of the invention is to lower down the activity of grazer and to inhibit the growth of grazer (in particular ciliate) by adding quinine preparation. The activity of the said grazer lowers down at lower concentration of the said compound and inhibition of the growth of the said grazer at higher concentration of

the said compound and thus, promoting the growth of algae cells for obtaining higher biomolecular content.
Another, embodiment of the invention is not affecting the biomolecular concentration of algae (in particular micro algae) while inhibiting the growth of the said grazer using the said compound.
Yet another, embodiment of the invention is to produce any one of the higher chains of biomolecules like carbohydrates, lipid or protein constituents.
Yet another, embodiment of the invention is to raise the over all process cost efficiency.
Further, embodiment is to produce continuous production of algae (in particular micro algae ) biomass for production of biofuel and bio-products.

EXAMPLE: 1
Micro algae particular Dunaliella species was cultivated in growth medium containing 2M NaCL 5mM MgS04, 0.75 mM KNO3, 0.2mM KH2PO4, 0.3 mM CaC12, 7μM MnC12, 5 μM EDTA, 2 μM FeCI3, 1 μM CUCI2: 1ΜM COCl2, 1 μM (NH4)6Mo7024, 1 μM ZnC12, 50mM NaHCO3 and 40mM Tris-Hcl (pH - 7.5 ). The media were sterilized at 121 °C for 20 min. Phosphates and carbon sources were autoclaved separately. Control set (medium without quinine preparation) along with experimental set (medium with quinine preparation) were carried out. Each analysis was performed in triplicates. Following data represents the mean values of the results.

Graph 1: PH of micro algae cultivation medium of without quinine (control) and with quinine (experimental)


Graph 2: Cell count during micro algae cultivation in medium without quinine (control) and with quinine (experimental)

Graph 3: Chlorophyll content of algae biomass during cultivation in medium without quinine (control) and with quinine (experimental)

Graph 4: TDS of a/gae cultivation medium medium cultivate in medium without quinine (control) and with quinine (experimental)

Fig. 1 Microscopic view of algae cultivate in medium without quinine (control) and with quinine (experimental)

WE CLAIM:
1. A methods for cultivation of contamination free algae (particular micro algae) cultivation.
2. The method of claim 1, where in medium containing quinine is ranging from 0.1 -50mg/L concentration of quinine preparation.
3. The method of claim 1, where in said cultivation methods could be open cultivation and closed cultivation.
4. The method of claim 3, where in said open cultivation methods could be open pond, open race way pond, open circular pond, open cultivation in any vessels of any shapes and types.
5. The method of claim 3, where in said close cultivation methods could be flask, close container, photobioreactor, or any close cultivation system.
6. The methods of claim 3, where in said phobioreactor could be of tubular, vertical, horizontal or any types with any cultivation capacity.
7. The method of claim 1. where in said cultivation medium comprises inorganic slats, further it comprises NaCl, MgS04, mM KNO3, M KH2P04, mM CaCI2, MnCI2; EDTA, FeCI3, CuC12, COCI2, (NH4)6MO7O24,. ZnCI2, NaHC03 and Tris-HCI.
8. The method of claim 1. where in said cultivation medium comprises inorganic salts, further the said in organic salt not only limited to as mention in claim 7, but also of any algae cultivation medium containing any inorganic or organic salt.
9. The method of claim 1, where in said micro algae is Dunaliella species. But the same methods could be applicable to any types of micro algae.
10. The method of claim I, where in said contaminant (grazer- particular ciliate) is reduced effectively, further same system could be from cultivation medium, further is equally effective for protozoans and bacteria.
11. Algal biomass generated by method of claim I could be used in preparation of biodiesel, food, feed and any industrial and non industrial use.
12. The method of claim 1 where in said cultivation medium does not affect growth requirement of algae (particularly micro algae) like pH, and TDS.

13. The method of claim 3 where in the cultivation medium is comprised of all type of water including fresh water, marine water, brackish water, sewage water, industrial and non industrial effluents, lagoon water.
14. A methods for cultivation of contamination free algae as herein described with reference to foregoing example.