The subject of the invention is a procedure for preserving soil bacteria, suitably to produce preserved soil-improving bio-fertilizers.

The market of alternative soil-improving and soil fertility restoring bio-products is growing rapidly, and the market competition is intense to develop products that are more efficient, can be stored and transported for longer periods, and are capable of restoring the ratio of nutritive materials in, and the fertility of, the soil. The state of the art includes the following solutions. Chinese patent descriptions Nos. CN101200385A and CN101200387A described an artificial fertilizer, where a solid carrier is covered with bacterium suspension, and then it is mixed with organic and inorganic components, and the mixture is fermented and dried subsequently. The presented artificial fertilizers include the mixing of well-defined microbes and organic waste, and a marketable product is produced from waste. Chinese patent document No. CN101913964A describes an artificial fertilizer, where a carbon carrier is covered with cultivated bacteria, the produced powder is closed in a capsule, and then it is mixed with organic components. In other words, it describes a capsule-based technology, where the capsule contains polymers that are turned into monomers by an auxiliary bacteria, and the soil bacteria use the monomers produced this way.

Japanese patent description No. JPH0641532A describes an artificial fertilizer, where various bacteria carried by various carriers are mixed with organic components. The described artificial fertilizer is a specialized product, which uses fungi to turn waste from the food industry into bio-fertilizer.

The solution described in Romanian patent description No. R0122676B1 is an artificial fertilizer, where various bacteria carried by various carriers are mixed with other nutritive materials, and the mixture is dried. The described procedure may be used with eukaryotes only, and it is not fixed to carbon.

Taiwanese patent description No. TW201439035A describes an artificial fertilizer. During the process, the fermented liquid is turned into a powder. Then, two kinds of materials are added, and activated carbon is not mentioned. The product can be stored for two years. It is used for bacteria, as well as eukaryote micro-organisms. Organic carbon sources are mixed to the cells to provide nutritive materials. This function, as it is claimed, is needed to activate the cells. On the other hand, this invention uses an inert carrier, and the metabolism of the cells is specifically activated before they are discharged. The invention described in Chinese publication document No. CN103964931 A utilizes existing waste material as bio-fertilizer.

Chinese patent document No. CN103992184A describes a specific artificial fertilizer product, which includes bacteria and carbon powder, but it can be stated in summary that it is produced for another intended use, and it includes less and other components.

Chinese patent description No. CN101429059A describes a bio-fertilizer enhanced with organic materials, which is supported through the mixed fermentation of various bacteria. The solution described in Korean patent document No. KR20130023001A is an artificial fertilizer, which includes various dried organic components, as well as carbon powder and bacteria; technically, it is a mix of organic waste materials improved and turned into a bio- fertilizer material.

The problem with bio-fertilizers described about and forming part of the state of the art is that their transport is rather expensive, as a thin and watery mass is transported, and they cannot be stored. The large volume also makes storage expensive and less feasible, and the product expire in a very short time.

This technology differs from the known solutions described above. The solution is a procedure that can be applied with prokaryotes, and is validated with three soil bacteria that belong to very different classes and have very different morphologic characteristics.

The purpose of the invention is to overcome the shortfalls of the known solutions, and to implement a procedure the use of which ensures the preservation of the inoculating culture of soil bacteria, economic transportation, simple handling, and fitness for long-term storage. Another goal is to achieve customizability to satisfy user needs by using various bacteria.

The inventive step is based on the recognition that an invention, which is more advantageous than the previous ones, may be created by executing the procedure according to claim 1.

In line with the desired purpose, the most general procedural form of the solution according to the invention may be realized according to claim 1. The various implementation forms are described in the sub-claims. In the course of applying the invention in a general manner, at least one bacterium tribe is cultivated in a cultivating dish and/or cultivating flask and/or fermenter, and the cultivated bacteria are moved into a mixing pot where they and are mixed. A distinctive feature of the application is that 15-200 g/L of activated carbon is added during slow stirring, and then the pH value of the mixture is changed by adding a solution, and then the mixture is divided into fermented liquid and a mixture of bacteria and carbon in a separating unit, and then the fermented liquid is removed, and the mixture of bacteria and carbon is rationed.

In another implementation form, one kind of bacterium tribe is cultivated in the cultivating dish and/or cultivating flask and/or fermenter before being moved into the mixing pot.

In another implementation form, one kind of bacterium tribe is cultivated at a time in the cultivating dish and/or cultivating flask and/or fermenter before being moved into the mixing pot, and the cultivation process is repeated according to the number of bacterium tribes to be cultivated, and then multiple bacterium tribes are moved into the mixing pot.

Another distinctive feature may be that, before portioning, the mix of carbon and bacteria is channelled from the separating unit to the drying unit, and drying is performed at 30 to 40°C for up to 6 hours, and a filter or centrifuge is used as a separating unit.

Another distinctive feature may be that granulated activated carbon is used as activated carbon, the size range of the granulated activated carbon is 0.2 to 7 mm, and that 35-200 g/L of granulated activated carbon is applied.

Another distinctive feature may be that the pH value in the mixing pot is set to fall within the 4 to 8.2 range.

In another implementation form, a mixed tank reactor or column reactor (air-lift) is used as fermenter.

In another implementation form, the solution used to change the pH level is a NaOH and/or HC1 and/or H ₂ S0 ₄ and/or NA ₂ C0 ₃ solution.

Another distinctive feature may be that cultivation is performed at a temperature between 25- 40°C.

The invention is presented in more detail using a drawing of a possible implementation form. Figure 1 shows the steps of the procedure. Figure 1 shows the main steps of the procedure. The bacteria are cultivated in the cultivating dish 1 and/or the cultivating flask 2. The cultivating dish 1 is suitably a Petri dish, i.e. a laboratory dish made of glass or plastic, which is suitable for cultivating cell cultures. Cultivation is performed on solid and/or liquid agar. Cultivation on liquid agar is suitably performed in an Erlenmeyer flask, while shaking. Using the culture in the flask as inoculum, it is possible to perform fermentation in larger volumes, which is done in a fermenter 3. The parameters applied are the same as the parameters applied during cultivation in a flask. The following possibilities are available for cultivation in a fermenter 3 : batch / fed batch / multiple fed batch. (Batch fermentation means fermentation in batches, Fed batch fermentation means fermentation in batches with feeding.) The fermenter 3 may be a column (air lift) reactor or a mixed tank reactor. Only one kind of bacterium is cultivated in the fermenter 3 at any given time. Then, activated carbon is fed, while stirring is performed in the mixing pot 4. In the next step, the pH level is set, while stirring is performed. Then, the carbon and bacteria mix is separated from the fermented liquid in the separating unit 5. Various technologies can be used for separation, taking into account the volume of the initial fermented liquid, the number of bacteria, and the rheology of the culture. It is possible to use centrifugal separation, in-depth filtration, tangential filtration, or cross-flow filtration. If filtration is used, the filtration device includes the following components: filter unit, output rubber hose for the filter unit, filtrate container with inlet and outlet stub, peristaltic pump, input rubber hose for the filter unit, filter unit rack. The bottom of the filter unit is perforated, and it includes a hollow into which the filters can be placed. During our work, we used a fibreglass filter. In order to increase the efficiency of filtration and to create a front layer, the captured granulated carbon was placed onto the fibreglass filters. Finally and optionally, drying is performed in the drying unit 6. This is necessary, if the water content of the bacterium product is not low enough.

Another implementation form is described below on the basis of experiments performed. The procedure according to the example includes the following steps. First, a dedicated agar is created for each bacterium tribe. The agar composition is calculated for 1 litre, distilled water is used as solvent.

Agar composition for B. subtilis Luria-Bertani:

SIGMA-ALDRICH LB Broth (Lennox) product is used. The agar includes trypton, yeast extract, and sodium chloride in the same as quantity trypton.

Agar composition for P. putida LB: Material Volume

peptone lO g/l yeast 5 g/l

NaCI lOg/l gar composition for S.coelicolor R5:

Material Volume saccharose 103 g/l glucose 10 g/l

L-prolin 3 g/l peptone o.i g/l

yeast 5 g/l

K ₂ S0 ₄ 0.25 g/l

MgCI ₂ *6 H ₂ 0 10.12 g/l

NaH ₂ P0 ₄ 5.73 g/l

KH ₂ P0 ₄ 5 g/l

CaCI ₂ H ₂ 0 2.58 g/l

Microelement solution for R5: for 100 ml

The optimal pH level of the agars was calculated as pH=7, which level was set by adding 3 M NaOH solution to the LB agars, or 5 M NaOH solution to the R2YE (R5) agar. In the course of producing a solid nutrient substratum, 2% agar is added to the LB nutrient substratum and 2.2% agar is added to the R2YE (R5) nutrient substratum per 100 ml.

In the next step, the cultivation of bacteria is performed. According to our example, the cultivation of the Bacillus subtilis is performed as follows: The B. subtilis tribe is taken from a culture, and it was grown at 37 °C on a liquid LB nutrient substratum shaken at 200 rpm in an Erlenmeyer flask, closed with a flask lid. Similarly to P. putida, this is a quickly growing tribe, which grows within 6 to 7 hours and reaches the peak of the exponential stage by the 8th hour. If grown on a solid nutrient substratum, 16 to 24 hours is enough at 37 °C.

Cultivation of the Pseudomonas putida tribe: The P. putida F 1 tribe is taken from a culture, and it was grown at 28 °C on a liquid LB nutrient substratum shaken in a shaker at 200 rpm in an Erlenmeyer flask, closed with a flask lid. Since it is a quickly growing tribe, it can grow within 4 to 5 hours, and it reaches the peak of the exponential stage by the 7th hour. If grown on a solid nutrient substratum, 12 to 18 hours is enough at 28-30 °C in the incubator.

Cultivation of the Streptomyces coelicolor tribe: The S.coelicolor tribe was always cultivated on a liquid R2YE (R5) nutrient substratum at 30 °C, which was shaken in Erlenmeyer flasks at 250 rpm to stimulate growth. Two methods were used to cultivate the tribe. On the one hand, the tribe was grown by using two shots of a culture taken from a solid nutrient substratum, and, on the other hand, a spore suspension was produced. We were able to collect approximately 1 to 2 ml of spore suspension in a Petri dish, which was enough to inoculate 1 to 2 flask, as 1 ml spore suspension was added to 100 ml nutrient substratum.

Using the culture in the flask as inoculum, it is possible to perform fermentation in larger volumes, which is done in a fermenter. The parameters applied are the same as the parameters applied during cultivation in a flask. As for dosage, using batch or fed-batch cultivation may be an alternative method, it depends on the expected yield or the specific bacterium used. As for the reactor, it can be a mixed reactor, or an external or internal air-lift. The three test soil bacteria are the Gram-negative Pseudomonas putida, the Gram-positive Bacillus subtilis, and the Gram-positive threaded (Actinomycetales) bacerium Streptomyces coelicolor. These are common soil bacteria that cover a broad spectrum for physiological and morphological purposes. The bacterium tribes described below are only used as examples; their morphology is diverse enough to allow our procedure to be considered a universal procedure.

Description of and optimal pH for the pseudomonas putida

It belongs to the Bacteria domain, the Protobacteria tribe, the Pseudomonadales order, and the Pseudomonas genus. P. putida is a rod-shaped, Gram-negative, non-spore bacterium that uses cilia to move, and it can be found in most soils and waters. It is sensitive to environmental changes, and can respond to such changes by changing its location. In addition to standard metabolic paths, it also uses alternative bio-chemical solutions to break down carbon structures (e.g. the Entner-Doudoroff pathway). The cellular membrane composition and lipid structure of P. putida depends on the testing environment: the sebacic acid saturation, the production of cyclopropane sebacic acids, and cis-trans isomerism can vary between wide limits. The optimal temperature is 25 to 30 °C, it can be easily isolated, and it can tolerate a wide range of pH levels (pH 3-9), but optimal growth is achieved at pH 7. It can break down various aromatic compounds in an aerobic manner, including organic solvents, such as toluol, and it is often used for bioremediation. P. putida is capable of forming cultures on the roots of plants, establishing a mutually close relationship between the plant and the bacteria. The surface of roots makes it possible for the bacteria to grow in the rhizosphere, as it provides nutritive materials for the bacteria. In return, the P. putida induces growth in the plants and protects them against pathogens. Since it can stimulate the growth of plants efficiently, it can be used in bio-fertilizers, bio-composites, and to restore the nutritive material content of the soil.

The P. putida bacteria were grown at a pH=7 level. However, we were able to strengthen the relationship in an acidic pH range, instead of a neutral range. The ideal pH range is the pH=4.5- 5 range, where we were able to reduce the period required for adhesion by approximately half, when using carbon powder. The experiment was performed by growing the bacteria in a given nutritive environment, the samples were shaken with an appropriate volume of carbon, depending on the carbon, and then we measured the pH level, which was between pH 8 and 9. The, it was reduced to the desired range, thereby improving the adhesion. Electrostatic energy reduces due to the pH level change, and the van der Waals bonds exceed the intensity of electrostatic interaction, so the absorption increases. As carbon has a positive charge and the bacteria have a negative charge, the electrostatic interaction between them is advantageous. It was also proven that the bacteria adhere to carbon with higher ash content better, as well as to hydrophobic surfaces and carbons with large macro-porous volume. Due to the latter factors, granulated activated carbon is more advantageous, since it has a larger macro-porpous volume and ash content.

Description of bacillus subtilis

It belongs to the Bacteria domain, the Firmicutes tribe, and the Bacillales. It was named Vibrio subtilis originally in 1835, but it was renamed to B. subtilis in 1872. B. subtilis cells are rod shaped, Gram positive, non-pathogenetic bacteria, and they can be found in the soil, on the roots of plants, and the intestines of mammals. The Bacilli grow in a mesophillic temperature range the best (25 to 35 °C), the pH optimum is between pH 7 and 8. To survive, B. subtilis produces stress resistant endospores. Endospores are extremely resistant dormant cells, their role is to survive under disadvantageous circumstances. The cytoplasm, DNA, and ribosome is located in the middle of the endospore, and they are surrounded by an external layer, which is enclosed by a non-permeable rigid shell. Endospores do not have metabolism. They are capable of surviving under extreme physical and chemical circumstances, for example strong UV or gamma radiation, solvents, germicides, heat, pressure, or dehydration. Carbon, nitrogen, or phosphor limitation can trigger the sporulation process, but the process must be started before all nutritive materials are depleted. B. subtilis can exist as an independent cell, and it can form cell chains as well. It produces surfactin, which is a cyclic lipopeptide with exceptional surfactant power, a kind of antibiotics. Thus, the bacterium has antibacterial and antifungal effect. Due to this characteristic, they can be used in soil improving materials and bio-fertilizers. Description of Streptomyces coelicolor

It falls in the Bacteria domain, the Actinomycetales, and the Streptomycetaceae genus. S. coelicolor is a threaded, Gram positive bacterium. It forms differentiated colonies based on branching mycelium networks, which show morphological similarities to (eukaryote) threaded fungi (hence their old name: actinomyces). They live in the soil and pursue a chemo- organotrophic way of life: the characteristic odour of soil is a result of the activities they perform in the course of breaking down organic materials. The colour of air mycelia is grey- yellow. They produce pigments depending on the pH level, so they are excellent pH indicators. Blue-purple colour indicates an alkaline environment, and red indicates acidity. They produce antibiotics, immunosuppressants, and anti-tumour materials, which can be used in healthcare. S. coelicolor is one of the most known prokaryote model organism. The surface of the mycelium is hydrophile, so they can stick well to wet surfaces and each other. In a stationary condition, they grow a protective cell layer, so they can survive longer. Older hyphas are thicker than more recent ones. Air mycelia have a fibrillated external layer, which makes the external surface hydrophobic, thereby breaking the surface tension. Sporulation starts when the threading begins on the air mycelia at the septum; this is where spores are produced. The round spores form chains. S. coelicolor also participates in the circulation of nitrogen. The pH optimum for Gram-positive bacteria (Bacillus subtilis and Streptomyces coelicolor) Capturing the Gram-positive B. subtilis and S. coelicolor bacteria is less complicated, than capturing the P. putida bacteria. During our experiments, the microbes were grown optimally at a pH = 7 level. The capturing processes were also performed in this pH range, with the difference that the charge of the carbon surface changed somewhat during the first 20 minutes of adhesion, which is a consequence of the emergence of a biofilm, which changes the charge density. It increases the negative charges mostly, which increase the absorption capacity for positively charged components. The hydrophobic characteristic is also important. Initially, the interaction is weak and reversible, especially due to the electrostatic repelling force between the negative surfaces. After overcoming the initial difficulties, the microbes quickly establish less reversible connections using extracellular polymers, which helps the maintenance of the biofilm. The bond depends on the number of activated carbon bonding points, as well as on the way how the bonding point of the carbon meets the external layer of the bacterium. The bond reduces the porosity of the carbon, and the charge density also changes. After bonding, the carbon has a negative charge, and it increases the carbon's capacity for positively charged species. Basically, it was not necessary to apply any change to the pH range, since the adhesive processes run quickly on their own, but we were still able to make some changes in the processes. The experiment was performed by growing the bacteria in a given nutritive environment, the samples were shaken with an appropriate volume of carbon, depending on the carbon, and then we measured the pH level, which was between pH 8.5 and 9, once the end of the exponential phase was reached. Then, acidity was restored to pH 7, which increased the adhesion of still active cells. This step is not necessary when using granulated activated carbon, because adhesion was quickly and efficiently completed due to the ash content and the macro- porous volume.

In our example, growing was followed by the determination of the cell volume, using lactophenol cotton blue after the adhesion. The culture medium was inoculated using a respective cell suspension. Once the culture was grown, a sample was taken, and the cell count was determined. Thus, it was possible to know the approximate cell volume that needed to stick to the carbon surface. The appropriate carbon volume was added, and the samples were shaken for 2 days. The carbon samples were filtered, and the filtrate was examined to determine the volume of uncaptured cells. Then, another cell suspension was made from the captured material. 180 μΐ distilled water was added to 20 μΐ thin carbonated sample, and a few drops of lactophenol cotton blue solution was added to the samples to improve visibility. 20 μΐ thinned sample was moved into a Biirker chamber, and the volume of captured cells was examined. The grown cell volumes were compared to the cell volumes in the filtered material and the carbon cell suspension, and a percentage value was determined, which reflected the actual situation approximately.

Then, the viability of the bacteria was examined. When growing micro-organisms, you must know if the cells you are working with are in the process of growing, decaying, or possibly dying. There is a fluorochrome compound, acridine orange (3,6 - bis[methylamino] acridine; AO), which can be used easily to test the vitality of micro-organisms. AO is one of the oldest fluorochrome paints, which can connect to polynucleotides. AO is a compound that contains a triple aromatic ring, which produces different excitation and emission wave length maximums, depending on if it is connected to a single or dual threaded polynucleotide. In the first scenario, the excitation maximum is 450 nm, and the emission is 650 nm. Actively growing cells have a high messenger RNA (mRNA) level, which appears in an orange-red colour, while the material fluoresce in green when the mRNA level is low. Consequently, when a cell is painted with acridine orange, the cytoplasm has a red shade (due to the RNA units in it), while the nucleus appears in a green colour. It must be noted that fluorochrome, in addition to being able to connect to nucleic acids, can also accumulate in strongly acidic cell organelles (e.g. lisosomas, endosomas). For this reason, it is also suitable to plaint prokaryote and eukaryote cells. However, in comparison to fungi, fluorescence appears the other way around. In bacteria, strong and viable cells fluoresce in orange-red colour, and weaker cells fluoresce in green; in fungi, strong cells fluoresce in green, and less vital cells fluoresce in a reddish colour. The samples were examined under UV light using fluorescent microscopes in the already mentioned wave length range.

Viability examination was performed do decide if the bacteria captured in the activated carbon remained viable after drying, and if they can be revived by being solved in a fresh nutrient solvent after several months in storage.

Then, the volume of activated carbon was optimized, and cell connections were established. The volume of carbon powder was set simply by trial series. Carbon powder was added gradually to 100 ml of fermented liquid, and the and the mix was observed under microscope to determine the optimal volume that is capable of picking up cells in large volumes from the nutritive environment, as well as the volume where additional adsorption is not achieved any more. Adsorption was stimulated using a shaking table. We also noted that Gram-negative bacteria are more difficult to capture. The volume of granulates was set using a method that is similar to the one used for carbon powder. Interesting observations were made during the volume related experiments. When adding GAC (granulated activated carbon), the carbon begin to dissolve in the nutritive environment, meaning that small carbon particles were dissolved from the granulate and C0 ₂ gas was developed. A shaker table was used to facilitate the creation of bonds, and the shaking accelerated the dissolution as well. This is an advantageous factor for us, because the bacterium cells could stick both to the inside of the granulate and to the carbon particles that dissolved from the granulate, meaning that a considerably larger volume of cells was captured eventually.

Steps of examining the binding of cells:

1. The tribes are grown under appropriate conditions.

2. A scale with taring function was used to measure the optimal volume of 4 g PAC

(carbon powder) or 10 g GAC into 100 ml of fermented liquid.

3. The pH level was optimized as necessary to ensure better and faster bonding.

4. Depending on the tribe, the carbon and bacterium cultures were shaken with a specific speed (rpm).

5. Sampling after 2 days of shaking.

6. 180 μΐ distilled water was added to 20 μΐ carbonated sample.

7. 20 μΐ thinned sample was taken and moved into a Biirker chamber to examine the captured cells.

Both Gram-negative and Gram-positive bacteria are captured on the granulates in large volumes. The bonds formed by the bacteria are influenced by various factors, such as function groups that are formed on the surface of carbon; the biofilm formed by the bacteria, which change the charge of the carbon; metal-oxides and electrolytes that give the carbon a negative charge; macro-porosity and ash content; and any pH change, depending on if Gram-negative or Gram-positive cells are to be captured. 3 M NaOH and 1 M HC1 solutions are used to set the pH level, and universal indicator paper is used for testing.

Then, it is possible to measure the dry volume of cells. The dry cell weight (DCW) is determined from a 100 ml sample from the liquid culture. The biomass and carbon aggregate is collected with a pre-measured paper filter and vacuum filter, and the filter is dried at 80 °C. This makes it easier to measure the captured cell volume.

If captured with carbon powder, measurement is performed as follows: The 100 ml flasks with carbon were shaken for 2 days, and the weight of the paper filter was measured. The samples were filtered using a vacuum pump and glass filter. The filtered samples were placed into a drying cabinet at 80 °C for 4 hours and they were dried until their weight became constant, and then they were placed into an exicator to cool. Finally, the weight of the sample was measured, and the weight of the paper filter and the carbon was deducted.

If captured with a granulate, measurement is performed as follows: The 100 ml flasks with carbon were shaken for 2 days, and the weight of the paper filter was measured. The samples were filtered with a Biichner funnel, and the filtrate was filtered further using a vacuum pump and glass filter. The filtered samples were placed into a drying cabinet at 80 °C for 4 hours and they were dried until their weight became constant, and then they were placed into an exicator to cool.

Then, the bacteria were separated from the fermented liquid, for example by using a filtration device. The bottom of the filter unit of the filtration device we used is perforated, and it includes a hollow into which the filters can be placed. During our work, we used a fibreglass filter. In order to increase the efficiency of filtration and to create a front layer, the captured granulated carbon was placed onto the fibreglass filters. By applying layers of various filters, it is possible to determine which bacterium tribe should be filtered first. The order is as follows: 1. S. coelicolor 2. B. subtilis 3. P. putida. The order starts with the largest cells and continues with smaller ones. The filtration of large and threaded bacteria is advantageous for filtration, as, due to its morphology, it creates an ideal surface for filtering other bacteria. Large mycelium can clog the pores of the filter, so smaller bacteria are easier to catch, and a front layer is formed more quickly. Various technologies can be used for separation, taking into account the volume of the initial fermented liquid, the number of bacteria, and the rheology of the culture. It is possible to use centrifugal separation, in-depth filtration, tangential filtration, or cross-flow filtration. If the applied filtration technology is not enough the produce a biomass and carbon mix with low enough water content, the technology also includes an optional drying step. Drying is performed at 30 to 40 °C for up to 6 hours.

In summary, the bindings of bacteria are determined by the following factors:

functional groups that develop on the surface of carbon

· biofilm created by the bacteria, which changes the charge distribution of the carbon

metal oxides and electrolytes that create a negative charge on the carbon surface macro-porosity and ash content

acidity, alkalinity, depending on if Gram-negative or Gram-positive cells are to be captured For the procedure presented regarding the implementation form, the optimal carbon volumes are as follows.

Granulated activated carbon (GAC): 100 g/ L, range that was tested and found suitable: 35-200 g L

Carbon powder (PAC): 40 g/ L, range that was tested and found suitable: 15-200 g/L

The optimal binding conditions for tested bacteria are as follows:

P.putida: 200 rpm, 28 °C, pH level between pH= 4.5-5, range that was tested and found suitable: 4-6

B.subtilis: 200 rpm, 37 °C, pH level between pH= 7-8, range that was tested and found suitable: 6.2-8.2

S.coelicolor: 250 rpm, 30 °C, pH level between pH= 7-8, range that was tested and found suitable: 4-8.2

Dry cell weight measurement can be used to determine that the following volumes of bacteria was captured in the product produced using the described process.

Carbon powder:

1. Pseudomons putida: 0.5-1 g

2. Bacillus subtils: 1-2 g/L

3. Streptomyces coelicolor: 3-5 g/L

G ra nulated ca rbon :

1. Pseudomons putida: 3-6 g

2. Bacillus subtils: 3-7 g

3. Streptomyces coelicolor: 5-8 g

After using a Biirker chamber to count the cells, it was established that an average of 2 to 35 cells are captured by a single carbon particle, depending on the size.

The procedure described above has numerous advantages. An advantage of the procedure is that it provides a universal solution for preserving any soil bacterium (or a mixture thereof), and the preserved product remains viable for at least 6 months. The procedure is a unique preservation method, which reduces the volume of water to be transported, and increases the dry matter content to almost 100%. Another advantage is that the procedure is simple and fast. Another advantage is its variability, as the typical microbes and their proportion can be adjusted according to the needs of the planned plants and local soil conditions. In other words, the composition can be adjusted and optimized using scientific methods. An important advantage is that the produced product can be transported at room temperature and, consequently, at reasonable cost. The produced product is a viable microbe flora bound to an activated carbon carrier, which is a dry powder and can be handled easily, and it can be released at the target area using, for example, a sower or an artificial fertilizer dispenser. It is not heavy as it does not contain water. Since it does not contain water, it can be transported and stored easily and at low cost. It improves the nutritive material content of the soil, and restores carbon sources. The procedure is universal and can be used with any suitable bacterium. In our example, a soil improving biofertilizer conserve was produced with three different bacteria, but the procedure can be used with only a single type of bacteria as well.

The field of application of the invention is the improvement of nutritive material content, the improvement of soil flora, the triggering of the reproduction of necessary soil bacteria, and the capturing of contaminants. In addition to the above examples, the invention can be implemented in other forms and with other manufacturing processes within the scope of protection.