WO2014120045A2

WO2014120045A2 - Method and product (options) of processing chlorella planktonic strain suspension, algolized compound feed-stuff, method of its production, method of introducing the derivative product in a watering system for birds or pigs

Info

Publication number: WO2014120045A2
Application number: PCT/RU2014/000046
Authority: WO
Inventors: Mikhail Vladislavovich KUNITSIN
Original assignee: Kunitsin Mikhail Vladislavovich
Priority date: 2013-02-01
Filing date: 2014-01-22
Publication date: 2014-08-07
Also published as: RU2538399C2; RU2013104225A; WO2014120045A3

Abstract

The invention belongs to bioengineering, namely, to a method of processing chlorella planktonic strain suspension. The method includes electroflotation, separation of the produced foam together with a small amount of suspension or cultural medium sufficient to ensure cell density 30 to 1000 mln/ml in the obtained chlorella biomass concentratee, removal of cultural medium. The cultural medium is concentrated. The method allows obtaining products in the form of chlorella biomass concentrate, cultural medium concentrate, and their mix. The method provides cost saving and raising the technological efficiency of chlorella use in medicine, cosmetology, animal and crop raising, and other areas.

Description

Method and product (options) of processing chlorella planktonic strain suspension, algolized compound feed-stuff, method of its production, method of introducing the derivative product in a watering system for birds or pigs.

The invention belongs to bioengineering, namely, to a method of processing chlorella planktonic strain suspension.

Globally, chlorella is used in three forms:

- dry (powder, tablets). Mainly, tablets as BAA for humans;

- paste;

- suspension.

Disadvantages of using dry chlorella: low digestibility of dry substance and heavy losses of biologically active substances in the course of drying and storing, i.e. value of the finished product becomes next lower than that of the primary product.

Paste starts spoiling within several hours due to considerable content of protein and fats, thus it is conserved using salt, chemical preservation agents or frozen.

Disadvantages of using suspension is the need of transporting large volumes of liquid (suspension), resulting in heavy transportation expenses and low technological effectiveness of feeding suspension to diverse animal species (designs of animal watering systems poorly fit or unfit for these purposes).

We know methods of producing Chlorella vulgaris strain suspension IGF No.C-111 and BIN using units (chlorella cultivators) of varied design. Herewith, the density of chlorella cells in the produced suspension range from several mln/ml to 10-15 mln/ml. The chlorella suspension of such cell density can be efficiently used in almost all lines of animal raising and crop raising (RF patents: No.2454067, No.2448453, No.2448457). The common disadvantages of using chlorella suspension are both the need of moving (transporting) large volumes of suspension and low technological effectiveness of using or impossibility to use suspension in certain lines, e.g. algolization of compound feed-stuff using chlorella.

We also know the method of feeding chlorella suspension to diverse animal species (Chlorella Suspension in Rations of Farm Livestock./ N.I. Bogdanov - 2^nd edition, revised and corrected. Volgograd, 2007-58 p.), according to which chlorella suspension, pursuant to the feeding norms for each animal species and groups, is included in the existing animal watering systems, mixed with feed-stuff immediately prior to feeding, or introduced in the pellet composition when producing pelleted compound feed-stuff.

The above method is technologically ineffective, labor-consuming and requires substantial expenses for transporting large volumes of chlorella suspension. It is impossible to materially increase the protein percentage in feed-stuff and affect the pellet durability resulting of adding chlorella suspension due to low concentration of chlorella cells in the commercial suspension and limited percentage of suspension introduction into compound feed-stuff, caused by the raise in its humidity (not more than 3%).

We know the method of feeding animals with pastelike chlorella biomass (M.Y. Salnikova. Chlorella - New Type of Feed-Stuff / M.Y. Salnikova. - M.: Kolos, 1977. - 95 p.), herewith, the method requires preservation of pastelike chlorella using diverse preservation agents, salt, or freezing.

We know the method of feeding animals with dry chlorella biomass powder (M.Y. Salnikova. New Type of Feed-Stuff / M.Y. Salnikova. - M.: Kolos, 1977. - 95 p. and A.M. Muzafarov. Cultivation and Application of Microalgae / A.M. Muzafarov, T.T. Taubaev. - Tashkent: Fan UzSSR, 1984. - 136 p.), which is too costly and inefficient due to low digestibility of dry chlorella biomass with animals.

The suggested invention allows eliminating the disadvantages of chlorella use and efficient processing of chlorella suspension with obtaining 3 products in a single production process. Besides, the method provides cost saving and raising the technological efficiency of chlorella use in the form of derivative products (concentrates) in medicine, cosmetology, animal and crop raising, and other areas.

To attain the technical goal, at least 10-fold (30-50-fold preferred) thickening of chlorella planktonic strain suspension has been suggested.

The original commercial suspension, produced using dedicated units (chlorella cultivators), has a low cell density. Obtaining a higher cell density during the photosynthesis or chlorella cultivation process is irrational and associated with using a more intense technology, sophisticated and expensive units, and significant appreciation of the final product. The suggested method allows considerable, up to 100-fold, increase of the cell concentration in suspension without breaking the initial balance of chlorella metabolites in cultural medium or destroying microalgae cells.

Processing of chlorella planktonic strain suspension can be carried out in three stages, herewith, biologically valuable products, which differ from each other in properties and applications, will be obtained at each of these stages.

At the first stage, two products will be obtained by electroflotation of the original chlorella suspension:

- concentrated chlorella cell biomass (CCB), which is a concentrate of living chlorella cells in cultural medium; - cleaned chlorella cultural medium (CCM), which is the chlorella cultural medium cleaned off chlorella microalgae cells.

At the second stage, using the reverse osmosis or ultrafiltration method, 5- to 100-fold thickening of cleaned CCM, obtained at the first stage, and deriving concentrated chlorella cultural medium (CCCM) are carried out.

At the third stage, CCB and CCCM are mixed together, and concentrated chlorella suspension (CC) is obtained.

The suggested method can be realized only with the planktonic strains having strongly marked bactericidal properties, e.g. with strains of Chlorella vulgaris IGF No.C-1 11 (SU 1751981, C 12N 1/12, 10.02.1997) and BIN (RU2263141 , C 12N 1/12 , 10.02.2005) or daughter chlorella strains (301670, C 12N 1/12, 19.05.2010) with high bactericidal properties, obtained on their basis. High bactericidal properties of planktonic strains protect microflora from outburst and prevent qualitative change of bacterial background even upon thickening of cells by dozens times against the original density. Chlorella cells remain in the same condition, as in the commercial suspension. Besides, CCCM is added to CC, i.e. concentration of antibiotic substances (Chlorellin) is dozens times higher. Still, basically, in CCB it also prevents cells from spoiling. Antibiotic Chlorellin does not suppress beneficial microflora of animal bowel - only pathogenic, hence it makes no harm to organism in contrast to all known antibiotics.

Chlorella planktonic strains' possession of high bactericidal properties provides considerable storage periods of derived products upon meeting normal conditions and does not require their preservation.

Detailed description of implementing the chlorella suspension processing method.

Stage 1.

According to the known method (SU No.1091890), the original chlorella suspension is subject to electroflotation; positively charged oxygen bubbles, resulting of water electrolysis, enable floating up of biomass to the surface and its concentration; then the biomass is removed to a receiver.

If solely foam is removed from the surface, a highly-concentrated chlorella cell biomass is obtained. Under standard conditions, the storage period of highly-concentrated biomass does not exceed one day due to its high concentration of proteins, fats, and carbohydrates. To extend the biomass storage period, it must be conserved by adding common salt, preservation agents or freezing.

We have found that upon adding chlorella cells to highly-concentrated biomass in a certain proportion sufficient to ensure the cell density 30 to 1000 mln ml in the obtained concentrate of chlorella biomass, cultural medium or chlorella suspension (having high bactericidal properties) it is possible to obtain a product - concentrated chlorella cell biomass (CCB), which can be stored already for not less than 30 days under standard conditions.

In practice, it is achieved owing to the fact that removal of foam from suspension surface upon electroflotation occurs simultaneously with a small amount of chlorella suspension, i.e. the foam collected not in the pure state but together with chlorella suspension. Chlorella suspension has high bactericidal properties and acts as a preservation agent even in case of 100-fold increase of the cell density against the original chlorella suspension.

The obtained cleaned chlorella cultural medium (CCM) is also a valuable biological product, which contains a wide range of dissolved biologically active substances - chlorella cell exometabolites (Chlorella, Physiologically Active Substances and Their Use. S.S. Melnikova, E.E. Manankina. Institute of Photobiology of AS BSSR, Minsk, 19 1).

Stage 2.

For the purposes of thickening cleared chlorella cultural medium (CCM) to the values 5 to 100, the well-known reverse osmosis method or ultrafiltration are used.

The reverse osmosis method is used for concentrating all the substances, contained in the cleared cultural medium (chlorella cell exometabolites), and separating pure solvent (water) from the solution, excluding a certain amount of monovalent ions Na, K, CI. Practically, reverse osmosis is reduced to thickening cleared chlorella cultural medium and obtaining the product of concentrated chlorella cultural medium (CCCM). Nobody has ever used such thickening method for chlorella cultural medium. Today, there is a wide diversity of reverse osmosis units. Concentrating is carried out, at least, by an order and, when needed, by 30-50 times. The storage period for CCCM is not less than 3 months.

Concentrating can be performed by ultrafiltration. Using a pump, pressurized cultural medium is filtered from within to the outside of UV module's hollow fibers. Cleaned water passes through porous walls of hollow fibers and it is further used as common clean water. Particles of chlorella cultural medium sized 0.01-0.1 μπι stay inside the UV module's hollow fibers. Concentrated chlorella cultural medium is accumulated in the receiver of thickened cultural medium resulting of fiber washing.

The ultrafiltration method is easier in terms of its execution, yet it allows concentrating only a particular range of chlorella cultural medium substances, the molecular sizes of which are 0.01 to 0.1 μ^ηι, what is a key disandavntage of this method. Stage 3.

CCCM and CCB are mixed together to obtain CC. Amounts of CCB and CCCM depend on the final product application, on the concentration (extent of thickening against the original) of CCB and CCCM. Mixing can be carried out in diverse concentrations for various applications. In some cases, it is reasonable to derive CC from one volume of the original suspension, i.e. both thickened CCB biomass and thickened CCCM are derived and mixed together afterwards. This being the case, we can talk of a product thickened against the original commercial suspension by several times (10, 15, 20, 25, 30, etc. times).

The storage period for CC is not less than 3 months under standard conditions.

The suggested method is cheap, simple, and reliable. The raise in the self-cost of processing one liter of chlorella suspension into concentrated CC will be not more than 10% of the proper commercial chlorella suspension production self-cost, and CCB - not more than 5%.

Characteristics of the products derived from chlorella suspension processing.

The suggested method does not disturb the structure and integrity of chlorella cells in derived CCB and CC products. Chlorella cells retain all of their properties, remain integral and able to reproduce, and perform photosynthesis throughout the entire storage period. Concentrated chlorella cultural medium (CCCM) retains all the properties of the original cleared cultural medium and the balance of chlorella exometabolites.

CCB product is a concentrate of chlorella cell biomass with cell density 30 to 1000 mln/ml, depending on the required value of thickening (concentration).

CCCM product is a cultural medium, which is left after separation of microalgae cells, concentrated by 5-100 times.

Concentrated chlorella suspension (CC) is a valuable biologically active product, derived by mixing the aforesaid products in efficient amounts.

Application of the products derived from chlorella planktonic strain processing.

Each of the derived products has its high biological value and various applications.

In case of applying chlorella suspension in crop raising, higher plants (their root systems or aboveground portions) use only chlorella cell exometabolites, dissolved in cultural medium. Herewith, the chlorella cells are not digested by higher plants and are, in fact, unnecessary ballast. Alongside with that, losses of valuable biological material - chlorella cell biomass - occur. Hence, in crop raising it is much more practical to use thickened chlorella cultural medium (CCCM) instead of the commercial chlorella suspension, and the obtained thickened chlorella cell biomass should be better used in other lines, e.g. in animal raising, medicine, cosmetology, etc. CCCM can be widely and efficiently used in crop raising as a growth-stimulating substance, immunomodulator, and supplementary feed for plants in order to increase yielding capacity of diverse cereals, vegetable and flower crops.

One of the essential lines of CCB and CC use is algolization of compound feed-stuff.

Algolization of compound feed-stuff with the derived products has the following advantages.

1. Increase in pellet durability. It is a vital parameter of pelleted compound feed-stuff. Besides, depending on the product thickness degree (thickening factor), pellet durability can be altered within widest ranges. They can be made almost "glassy", i.e. very solid, but such durability will reduce digestibility of compound feed-stuff.

Durability of compound feed-stuff is characterized by crumb formation that is determined according to GOST 28497-90.

There is GOST 51899-2002: Pelleted Compound Feed-Stuffs. General Specifications.

For example, pellet crumb formation for diverse animal species is set forth therein:

- for farm livestock - 22%

- for rabbit, nutria, and fur-producing animals - 8%

- for fish - 5%

- for horse - 7%.

Using concentrated products when producing compound feed-stuff, i.e. algolization of compound feed-stuff with CCB and CC products may lower the crumb formation percentage to 1% and below, depending on the concentrations of products. The amount of introduced products remains as 2-3% of the algolized compound feed-stuff mass with chlorella cell density 50 to 500 mln/ml.

In case of standard production of pelleted compound feed-stuff, a great focus is on the so- called coupling substances, which are introduced for both increasing pellet durability and cutting steam and energy consumption, productivity gain. Most commonly, liquid products, such as fat, hydrol, molasses, etc. and powders - bentonites, - are used as such substances. Some of the above substances enhance nutritional value of compound feed-stuffs (fat, molasses), enrich compound feed-stuff with microelements (bentonites). The amount of added coupling substances is normally small - up to 3 %. However, it is recommended to introduce large amounts of fat, up to 6%, to recipes of certain compound feed-stuffs for bird. Fat, introduced in the amount over 3%, ceases to be a coupling component. Moreover, upon introducing a large amount of fat, press performance and pellet durability (pellets show considerable loss of their durability) are reduced. This being the case, it is possible to apply other coupling substances, mostly bentonite for instance, what allows improving the process of compound feed-stuff pelleting. Nevertheless, introduction of concentrated products with chlorella is substantially more efficient and allows reducing the pellet crumb formation percentage almost by an order. When introducing highly- concentrated products in compound feed-stuff, the percentage of introduced protein also grows, what enhances the nutritional value of algolized compound feed-stuff materially.

Raise in pellet durability has a direct impact on increase of the press performance factor.

The press performance factor is a relation of integral pellet quality to the overall product, obtained after pressing. The higher pellet durability is, the higher the performance factor. Pellet durability is an important indicator of their quality. If pellets are insufficiently solid, they are destroyed during transportation, loading in bunkers, storage, handling.

For example, the compound feed-stuff for broiler chicken contains lots of fat and protein and little cellulose, so a concentration at least 5 times higher against the commercial suspension is needed to enhance the pellet solidity.

Pellets 0 2...3 mm are required for young birds, though producing such pellets is unreasonable due to abrupt cut in the press performance and increase in the specific energy consumption. The pelleting process is unstable during production of such pellets, matrix dies become clogged with compound feed-stuff. For example, if a press switches to production of pellets 0 2.4 mm instead of 4.7 mm, its performance drops by, roughly, four times and the specific energy consumption grows more than twice. Thus, fine pelleted compound feed-stuffs are produced by crushing pellets 0 4.7...9.7 mm with further calibration in sieves of specific sizes. So, for chicken and broilers aged 1 to 30 days, the remains on a sieve with mesh 0 3 mm must not exceed 20%, and throughs of a sieve with mesh 0 1 mm must not exceed 18%.

Pellets are crushed so as to obtain the minimum of fine fraction, which must be re- pelleted. The amount of such fraction must not exceed 30%, and the middlings extraction must be not less than 70%.

The suggested method for algolization of compound feed-stuff with thickened CCB and CC chlorella biomass allows substantial increasing of durability and lowering the crumb formation of the pellets used to obtain middlings. Due to cutting crumb formation during production of middlings for chicken and broilers aged 1 to 30 days, the throughs of a sieve with mesh 0 1 mm are not more than 2%, and the middlings extraction is not less than 95%.

Such effect is achieved upon algolization of compound feed-stuff with thickened CCB and CC chlorella biomass in the amount 3% of the compound feed-stuff mass with the CCB and CC thickening factor 5 and higher. Thickened CCB and CC chlorella biomass is delivered directly to the mixer preceding the pelletizer.

2. Increase in the nutritional value of compound feed-stuff.

Recipes of compound feed-stuffs are intentionally elaborated for age- and industry branch-related animal group (calves, dairy cows, feeder livestock); compound feed-stuff for birds can be starting, fattening, and for layers.

The nutritional value of compound feed-stuffs is determined by calculation based on the relevant nutritional value tables broken by three key indicators: content of feed units in 100 kg of compound feed-stuff; content of digestible protein (in grams per 1 feed unit); content of cellulose (in %).

Changing the thickening factor (concentration) of chlorella products can alter the nutritional value of algolized compound feed-stuff pursuant to recipes. Taking into account that the chlorella cell density in these products is high enough, and the protein content in an absolutely dry chlorella substance reaches 50-60%, it is fair to say this algolization method will be of great importance for the compound feed-stuff industry. It is common knowledge that the chlorella protein is valuable owing to both its high content in an absolutely dry chlorella substance and well-balanced and full composition of indispensable aminoacids in the chlorella protein. We have cheaply obtained a very valuable product that is easy to include in the processing chain of compound feed-stuff production. Herewith, the compound feed-stuff composition is maximally enriched with biologically active substances of natural origin, i.e. natural plant. Hence, we receive both the high economic outcome and environmentally safe products having high consumer properties.

Globally, nobody has ever included such product in compound feed-stuff - it used to be only the commercial chlorella suspension. As it has low cell density (about 10 mln/ml) and the percentage of its inclusion in compound feed-stuff is 3-5%, so its limited potential and, accordingly, effect are evident.

3. Reduction of plate count and extension of the algolized compound feed-stuff storage period.

Storing extensive amount of the processed product (compound feed-stuff being such) is associated with its oxidation, nutritional value loss, etc. Upon failure to observe the storage regulations, fats (triglycerides), plant oils, and products, containing a great amount of fat (feeding fat; meat meal, meat-and-bone meal, and fish meal), which are widely used in compound feed-stuffs as an energy resource, oxidize. Their oxidation both has a negative impact on feed-stuff taste and reduces its calorific value. The process of fat and plant oil oxidation is a multiple-factor process and depends on temperature (the higher temperature is, the faster oxidation), humidity, availability of oxygen, trace metals, light impact, etc.

The longer the feed-stuff storage period is, specifically under excessive temperature and humidity conditions, the higher the risk of the losses, caused by its spoiling by mold fungi.

Spoiled compound feed-stuff is specifically dangerous for layers and young stock during the initial period of its growth. Particles of spoiled feed-stuff further become polluters of fresh feed-stuff in feed cribs.

During their development, mold fungi and bacteria use the energy of compound feedstuff and feed materials. Development of fungi may also result in worsened taste of feed-stuff and its contamination with mycotoxins.

Aflatoxins, ochratoxin A, zearalenone, and trichothecenes are the most dangerous for human and animal health.

In the result of feeding compound feed-stuffs, containing feeding components (com, soy, etc.) contaminated with mycotoxins, poultry farmers suffer great losses. Damage occurs resulting of spontaneous mortality, decreasing liveweight gains, lowering egg-laying capacity, hatchability (embriotoxity), and resistance to diseases (coccidiosis, bacterial and viral infections).

Upon algolization with thickened CCB or CC products, a peculiar smooth film occurs on the surface of compound feed-stuff pellets. Accordingly, there is the effect of packing each separate compound feed-stuff pellet. Herewith, the smooth film (crust) impedes free penetration of microorganisms inside the pellet and oxygen. A double "preservation" effect occurs. High bactericidal properties of chlorella are active inside pellets, and the protecting coat - outside.

Owing to their high bactericidal properties, compound feed-stuff with chlorella included is less exposed to pathogenic microorganisms, what allows substantial prolongation of the compound feed-stuff useful life.

Method of algolized compound feed-stuff production.

Two methods of producing pelleted compound feed-stuffs, dry and wet, are applied at compound feed-stuff plants. According to the first method, dry bulk compound feed-stuffs are steamed before pressing, sometimes with introduction of liquid coupling additives (molasses, hydrol, fat, etc.). When using the wet method, hot water (70...80°C) in the amount, ensuring production of paste with humidity 30...35 %, is added to compound feed-stuff; then the paste is used for forming pellets that are further dried and chilled down.

Dry pelleting. A press with a rotating circular matrix is used for these purposes. Jets for supplying hot water or any coupling liquid and steam supply chambers are installed in the bladed mixer. Prepared compound feed-stuff is delivered to the pressing part of the pelletizer, which represents a rotating circular matrix and two pressing rollers (three in some deigns).

In the pressing part, the product is drawn in a V-shaped gap between the rotating matrix and the roller, driven by material (due to friction). When the product moves through the V- shaped gap, pressing of material occurs, what increases its density. At the moment, when compression stress exceeds resistance of the material, earlier pressed in the matrix dies, the product staying in the V-shaped gap starts being pressed into the dies and shifted towards them. The process is accompanied by pellets^' moving outside the external surface of the matrix that ends up at the moment of the die's passing through the section, where the height of the V-shaped gap is the lowest. Having passed through the dies, the product assumes the shape and sizes of pellets with the relevant density and durability. On exit from the matrix, the pellets are cut by two blades that can be moved closer or further, thus regulating the pellet length. The pellet durability can be changed by various preparation of the product, alteration of the gap between the roller and matrix.

The method of pelleted compound feed-stuff algolization is that the product of CCB or CC chlorella planktonic strain suspension processing, having chlorella cell density 50 to 500 mln/ml, is introduced in compound feed-stuff before its pelleting in a mixer or immediately upon pelleting (in a pelletizer) in the amount 2-3 wt % of the algolized compound feed-stuff mass; the maximum percent of the product input (amount) in compound feed-stuff is determined by maximally allowable humidity value of the finished compound feed-stuff.

CCB and CC application is not limited only by algolization of compound feed-stuff, they can be used in diverse areas: for algolization of substrates in mushroom (shiitake) raising, in manufacturing BAA for humans, in cosmetology, for enrichment of food products (bread, pasta, ice cream etc.), for algolization of water bodies, in animal raising for enriching feeding rations via watering systems and directly by adding them in feed-stuffs. CCCM can be used in growing vegetables and flowers in hydroponics and soil, any cereal crops - grain and pulses.

Thus, algolization of water bodies, carried out according to the method (Patent No.2370458, published on 20.10.2009), stipulates multiple introduction of a considerable chlorella suspension amount in a water body. Chlorella vulgaris IGF C-l l l strain suspension with transmission factor 1.4-6 in the amount 15-25 1 per 1 mln m³ of water, and in creek, draws and backwash locations on the basis 15-25 1/ha, have been used. This is explained by considerable expenses during its transportation. Transportation expenses are the key expensive item in algolization of water bodies. Besides, it is also impossible to obtain chlorella suspension at water body locations, as its cultivation is associated with meeting specific conditions (premises, equipment, technology, etc.).

Thus, using CCB products with high degrees of the original chlorella suspension thickening would contribute to significant reduction of expenses performing water body algolization operations.

The possibility of implementing the inventions is illustrated but not limited to the examples.

Chlorella vulgaris of the required density is obtained as follows.

Cultivation of the commercial suspension is performed in production units. For the purposes of launching production units, the required volume of chlorella mother culture is obtained in separate units - mother units. In mother units, Chlorella vulgaris (IGF No.C-11 1) is cultivated using an extensive technology. In a mother unit, the cell density increment is up to 15 mln cells in 1 ml. The derived chlorella mother culture is delivered to a production unit in the amount 20% of its overall volume. The initial cell density in the production unit suspension is about 3 mln cells in 1 ml. For three days of production unit operation, the commercial suspension density reaches the value of 10 mln cells in 1 ml. Afterwards, the ready-made suspension is discharged for further use, and the production unit is started up once again.

Example 1. Description of chlorella concentration (electro flotation) method:

Suspension of chlorella strain IGF No.C-11 1 with cell density 10 mln/ml in the amount 100 liters is subject to electroflotation. In the process of the suspension thickening, foam with a small amount of the suspension, which after settling represents a concentrate of chlorella cells in a small amount of the suspension, is collected. After setting-out during 30 minutes, a concentrated biomass product with the volume 1 liter is obtained. The chlorella cell density in the thickened product is 1000 mln ml. One liter of such CCB product contains about 70 grams of absolutely dry chlorella substance.

The product was being stored during 30 days at temperature 21 °C in dark premises. After storage, CCB has the following characteristics.

In terms of bacteriological properties, the derived product (CCB) is compliant with the veterinary requirements to bacteriological factors. The product does not contain pathogenic agents belonging to the groups of anaerobia, proteus, salmonella, enterococcus, enteropathogenic E.coli. Toxico-biological studies have shown that the product is non-toxic.

Example 2. Concentrating cultural medium using the reverse osmosis method. 95 liters of cleared chlorella cultural medium (CCM) have been obtained resulting of performing electroflotation of 100 liters of chlorella suspension.

Cleared chlorella cultural medium (CCM) is delivered to a reverse osmosis unit for thickening. The degree of concentrated chlorella cultural medium (CCCM) thickening depends on the reverse osmosis unit operation time and is determined by the relation of the original CCM volume to the obtained CCCM volume. So, during one hour of the reverse osmosis unit operation the volume of the obtained CCCM product was 9.5 liters. The degree of CCCM thickening, accordingly, was 10. After continuing operation during the next hour, the volume of the obtained CCCM product was 1 liter. The degree of CCCM thickening, accordingly, was 95. The product was being stored during 90 days at temperature 21 °C in dark premises. After storage, CCCM has the following characteristics.

In the result of mixing 1 liter of CCB and 1 liter of CCCM, concentrated chlorella suspension (CC) with the volume of 2 liters is obtained.

Thus, the degree of the CC product thickening is 50.

The product was being stored during 90 days at temperature 21 °C in dark premises. After storage, CC has the following characteristics.

Example 3. Concentrating cultural medium using ultrafiltration.

Cleared chlorella cultural medium (CCM) in the amount 95 liters is delivered to an ultrafiltration unit. The degree of concentrated cultural medium (CCCM) thickening depends on the ultrafiltration unit operation time and is determined by the relation of the original CCM volume to the obtained CCCM volume. So, during 5 minutes of the unit operation the volume of the obtained CCCM product was 4 liters. Accordingly, the degree of CCCM thickening was about 24.

In the result of mixing 1 liter of CCB and 1 liter of CCCM, concentrated chlorella suspension (CC) with the volume of 5 liters was obtained. Accordingly, the degree of the obtained CC product thickening was 20. The required degree of CC, CCCM, and CCB product thickening depends on the purposes and scope of their application and can vary within diverse ranges.

Example 4. Algolization of bird compound feed-stuff.

Using CCB or CC product for algolization of bird compound feed-stuff requires a higher degree of thickening. So, compound feed-stuff for broilers contains lots of fat and little cellulose, what has a negative impact on pellet durability. The experiment was carried out on pelleted compound feed-stuff for broilers BROILER ROST 1 1-24 days with crude fat content 7.1 1%. Upon introduction of CCB or CC product with thickening degree 5 in a mixer in the amount 3% of the compound feed-stuff mass, the pellet crumb formation dropped by 3 times against the control. The pellet crumb formation is determined as relation a:b, where - a number of the destroyed pellets, having passed through the sieve; b - initial number of the pellets.

Using CCB and CC products with a higher thickening degree for algolization of compound feed-stuff is needed for enhancing its nutritional value, complete protein content, volumes of exchange energy, vitamins, microelements, carotene, etc. In such cases, the degree of CCB and CC product thickening must be increased up to 50.

Example 5. Using the products, derived resulting of chlorella suspension processing, in feeding broilers.

Using CCB (CC) product for algolization of bird compound feed-stuff for broilers: PK-5- 1, PK-5-2 and PK-6, produced under STB 1842-2008. The algolization of compound feed-stuff was carried out on the basis CCB (CC) product volume 3% against the compound feed-stuff mass. CCB (CC) product has been put in a mixer prior to a pelletizer. The degree of CCB product thickening was 30.

CCCM is a valuable biological product, containing all water-soluble concentrated chlorella exometabolites, thus it must be also used in bird feeding. Concentrated chlorella cultural medium (CCCM) has been introduced into the bird watering system in the water treatment package via the medicator (doser). The medicator performs sampling of the concentrated product (CCCM), dosing against the required percentage composition, and mixing with water. The obtained solution is delivered further throughout the watering network. The medicator allows introducing a definite percentage of the dosed CCCM, 3% in our case, in water. According to this value, the optimum degree of CCCM thickening must be not less than 33. The CCCM volume is calculated relying on the recommended norms of chlorella suspension feeding for each animal and bird species. The mean norm of feeding the chlorella suspension with cell density 10 mln ml for broilers is around 20 ml/day per head. For one standard facility with livestock 30.0 thou, heads, the mean norm of chlorella suspension feeding is 600 1/day. The calculated degree of CCCM thickening is 33. Hence, the mean daily norm of CCCM for one facility with 30.0 thou, heads of livestock is 18.18 liters. In our examples, the norm of CCCM feeding has been evenly increased since the 5^th until the 38^th day of the chicken feeding from 5 to 40 liters per day proportionally to the broilers^' growth.

During the conducted experiment of feeding the broilers of Iza cross, the following results have been obtained. In the test group of the broilers, the raise in the average daily gains was 9.8% higher than in the control group. During the entire feeding period, the bird mortality lowered by 2.4 times and made 1.8%. The feed-stuff conversion dropped from 1.92 in the control group to 1.73 in the test group. The test bird's liver had a firmer texture and darker color. The cases of liver discarding in the test group were occasional. During the experiment, no antibiotics or other veterinary drugs, excluding mandatory vaccinations, were used in the test group. The taste of the test group's poultry meat and broth had a remarkable difference to the better as compared to the test group.

Example 6. Delivery of CC product in a bird watering system.

Under certain conditions of delivering CC in a watering system via a medicator, the effect of direct delivery of chlorella suspension in the watering system can be achieved. It is possible when the degree of CC product thickening corresponds to the medicator dosage percentage so that the chlorella cell density in the watering system is equal to 10 mln/ml or to the cell density in the original chlorella suspension, of which CC product was derived. In other words, should the degree of CC product thickening be 33.3 and the percentage of dosing CC product in the watering system via the medicator - 3%, the effect of direct inclusion of chlorella suspension in the bird watering system can be achieved, i.e. water is cut off and chlorella suspension is delivered directly in the system, as the work with chlorella suspension used to be carried out earlier. But before this used to result in the need of transporting large volumes of chlorella suspension to animal production units, herewith, installing dedicated receiver tanks for chlorella suspension and further cutting off water and delivering chlorella in each water treatment package of a bird raising facility. All this had to be supervised by a technician. Under the suggested method, CC can be automatically delivered via a medicator, same like vitamins, drugs, etc. delivered via a medicator.

Accordingly, we suggest introducing the concentrated product via a medicator that delivers dosed CC in the watering system, e.g. 3 or 5% of the water volume. Should the medicator be set for 3%, CC must be thickened by at least 33 times; should the medicator be set for 5%, CC must be thickened by at least 20 times to ensure the cell density of chlorella in drinking water 10-50 mln/ml, herewith we achieve the same effect as before when delivering the commercial chlorella suspension directly in the watering system with water cut off.

Upon implementing this method of introducing CC product in the watering system via a medicator, the equal results were obtained in practice, when chlorella suspension was included directly in the watering system pursuant to the watering norms for bird.

Example 7. Using CCB (CC) product in pig feeding.

CCB (CC) feeding was carried out via algolized compound feed-stuff on the basis 3% of the introduced product against the compound feed-stuff mass. The degree of CCB thickening was 20. CC was delivered in the watering system via a medicator. The coefficient of CC dosing by the medicator was 3%. According to this value, the optimum the degree of CC thickening must be not less than 33. The CC volume was calculated relying on the recommended norms of feeding the chlorella suspension with cell density 10 mln/ml for pigs. The daily norm of feeding chlorella suspension per head for breeding sows is 1 liter, for one month old weaned pigs - 200 ml, two months old - 300 ml respectively, etc. Accordingly, the mean daily norm of CC feeding for a breeding sow group of 90 heads will be 2.7 liters. The mean daily norm of CC feeding for a weaned pig group of 500 heads will be 3.0 liters.

The test breeding sow group in the amount of 90 heads was fed 3 liters of CC daily via a medicator in the watering system during the period since the sixth day of their being in season and insemination. Besides, the test group was receiving compound feed-stuff algolized with CCB on the basis 3% of the compound feed-stuff mass with the degree of CCB thickening 33.

Due to the increased breeding sows' milking capacity and milk fatness, suckling pigs had a high vital growth energy, differed from their herdmates in mobility and high immunity. Diseases and mortality of the young stock were not observed during the suckling period (two heads were stepped on by the breeding sows). Upon weaning at the age of 29 days, the mean weight of piglets was higher than that of the control group by 37%.

Besides, chlorella had a stimulating impact on the reproductive system of the breeding sows in the test group. Out of 90 heads in the post-weaning period during 6 days 87 breeding sows (96.6%) came in season, when during the same period only 65% breeding sows in the control group were in season.

CC was fed to the weaned pigs in the amount 500 heads via the watering system medicator every twenty-four hours in the amount 3 liters during the first months of the experiment and 4.5 liters - during the following months. The experiment period was 2 months since weaning. Besides, during the experiment period piglets were receiving algolized CCB compound feed-stuff in the proportion 3% of the compound feed-stuff mass with the degree of CCB thickening 10.

The piglet test group substantially surpassed the control group in development. The raise in weight gains was 27% in the first month and 34% in the second months of the experiment. The mortality level in the test group was lower than that in the control group by 4.7 times.

Example 8. Using CCB and CC products in keeping and feeding bovine cattle.

A group of milking cows in the amount 200 heads was fed CCB (CC) via a shredding mixer, regular mixer, wheeled distributor unit, in which complete feed of long hay, straw, silage, compound feed-stuff, food supplements, minerals for bovine cattle was prepared. The product was introduced in the mixer on the basis 0.3 liters per head. The product thickening factor was 30. The experiment period was 50 days. During one month, the milk productivity of the animals in the test group has increased by 19%. The fat content in the milk was higher than that for the control group by 0.15%. After completion of the experiment, the milk productivity in the test group remained at the same level (higher than that in the control group) for another two months, further observations were not performed.

The high cell concentration in the product, in contrast to the commercial chlorella suspension (density 10 mln/ml), had an insignificant impact on increasing humidity of the finished feed-stuff in the mixer and, simultaneously, allowed feeding the required amount of chlorella biomass to animals.

Example 9. Using CCC for stimulation, growth, and productivity of roses during cultivation in hydroponics.

CCCM application is associated with an increased level of chlorella use technological effectiveness in cultivation of roses in hydroponics, as the used volumes of liquid products, upon their inclusion in the plant feed system, can be reduced by dozens times. Besides, higher plant roots are unable to digest chlorella cells. CCCM contains a wide range of physiologically active substances: plant hormones (auxins), water-soluble vitamins, steroids, gibberellin-like substances, phenol compounds, and cytokinins. It is found that these substances are regulators of plant growth and development. Using CCCM as an ingredient in hydroponic solution entails plants' receipt of indispensable feed elements and growth-stimulating substances.

The amount of the introduced CCCM product is calculated as follows. There are 206 thousand flowers (roses) in a greenhouse of 3 ha. The nutrient medium volume for one watering is calculated based on the norm 60-70 ml per plant. Totally, around 14.5 tons of nutrient solution are spent for one watering. The required chlorella suspension volume is 10% of the nutrient medium volume, i.e. 1500 liters. When using CCCM with the thickening value 30, only 50 liters of CCCM are needed for one watering. Via a dedicated doser, the required CCCM volume is evenly distributed throughout the entire volume of the nutrient medium, delivered during one watering. CCCM is introduced in the feed system once daily.

The results of using CCCM are analogous to the results, obtained when using chlorella suspension.

Example 10. Using CCCM for stimulation, growth, and yielding capacity of cereal crops by the example of barley.

The experiment included the following options:

1. sowing of barley without CCCM treatment (control);

2. sowing of barley using seeds treated with CCCM;

CCCM had the degree of thickening 10. The CCCM volume was on the basis 30 liters of CCCM per 1 t of barley seed. The barley treatment was carried out via a seed dresser. The seed wetting degree was chosen so as to unable the increased seed humidity to have a substantial impact on impairment of the barley flowability during its sowing using a sowing-machine. The sowing rate was 150 kg/ha, the sowing acreage was 10 ha.

The experiment deliverables. The yielding capacity in the control field was 21.4 c/ha, the mean yielding capacity in the test field was 30.9 c/ha.

Claims

INVENTION FORMULA

1. Method of processing chlorella planktonic strain suspension, including:

a) electroflotation,

b) separation of the produced foam together with a small amount of suspension or cultural medium sufficient for ensuring cell density 30 to 1000 mln/ml in the obtained chlorella biomass concentratee,

c) removal of cultural medium.

2. Method under ex.1, characterized in that it also includes concentrating of the cultural medium, derived at stage c) by 5-100 times using the reverse osmosis method or ultrafiltration.

3. Product of chlorella planktonic strain suspension processing, being a concentrate of chlorella cell biomass with cell density 30 to 1000 mln ml.

4. Product of chlorella planktonic strain suspension processing, being a concentrate of cultural medium, derived using the method under ex.2

5. Product of chlorella planktonic strain suspension processing, being a mix of products under ex.3 and under ex.4 in the amount efficient for further use.

6. Product under ex.5, characterized in that the amounts of Product 3 and Product 4 correspond to the amount, derived of one volume of the original chlorella planktonic strain suspension.

7. Pelleted compound feed-stuff, algolized using the product under ex.3 in the amount 2-3 wt % at chlorella cell density 50 to 500 mln/ml.

8. Pelleted compound feed-stuff, algolized using the product under ex.5 in the amount 2-3 wt % at chlorella cell density 50 to 500 mln/ml.

9. Method of pelleted compound feed-stuff algolization, including introduction of the product of processing chlorella planktonic strain suspension under ex.3 or ex.5 in compound feed-stuff at chlorella cell density 50 to 500 mln/ml prior to pelleting in a mixer or, immediately when pelleting, in a pelletizer in the amount 2-3 wt%.

10. Method of direct delivery of the product under ex.5 in a watering system for birds or pigs, characterized in that the dosed product is delivered via s medicator in the amount ensuring chlorella cell density in water 10-50 mln/ml.