PL232671B1 - New Lactobacillus plantarum AMT12 strain and the composition containing the Lactobacillus plantarum AMT12 strain - Google Patents

Description

Description of the invention

The invention relates to a new Lactobacillus plantarum AMT12 strain and a composition containing the new Lactobacillus plantarum AMT12 strain.

Lactobacillus plantarum is classified as a lactic acid bacteria that has an important industrial function (Parente E., Ciocia F., Ricciardi A., Zotta T., Felis GE, Torriani S. 2010. Diversity of stress tolerance in Lactobacillus plantarum, Lactobacillus pentosus and Lactobacillus paraplantarum: a multivariate screening study. International Journal of Food Microbiology 144, 270-279). These bacteria are commonly found in fermented foods, olives, cheese, wine and silage (Tanganurat W., Quinquis B., Leelawatcharamas V., Bolotin A., 2009. Genotypic andphenotypiccharacterization of Lactobacillus plantarum strains isolated from Thai fermented fruits and vegetables, Journal Basic Microbiology 49, 377-385). Therefore, strains of this species, having probiotic properties, are used in the production of functional and therapeutic food and as potential oral live vaccines (Shah N.P., 2007. Functional cultures and health benefits. International Dairy Journal 17, 1262-1277).

The mechanism of action of probiotic bacteria is multifactorial and specific for individual strains (Tuohy K.M., Probert H.M., Smejkal C.W., Gibson G.R., 2003. Using probiotics and prebiotics to improve gut health. Drug Discovery 8, 692-700). One of the better-known modes of action of probiotics against pathogens is antagonism based on the release of bacteriostatic and / or bactericidal substances into the environment by probiotics described by Kesarcodi-Watson et al. (Kesar-codi-Watson A., Kaspar Hl., Lategan MJ, Gibson L., 2008. Probiotics in aquaculture: The need, principles andmechanisms of action and screeningprocesses. Aquaculture, 274, 1-14). The presence of probiotics in the intestine or on the surface of the host's skin is believed to inhibit the growth of potentially pathogenic bacteria (Verschuere L., Heang H., Criel G., Dafnis S., Sorgeloos P., Verstraete W., 2000. Protection of Artemia against the pathogenic effects of Vibrio proteolyticus CW8T2 by selected bacterial strains. Applied and Environmental Microbiology, 66, 1139-1146). This antibacterial effect is caused by individual or combined substances produced by bacteria, such as: antibiotics, bacteriocins, lysozyme, proteases, hydrogen peroxide, ammonia, diacetyl, siderophores, single organic acids (Verschuere L., Rombaut G., Sorgeloos P., Verstraete W., 2000. Probiotic bacteria as biological control agents in aquaculture. Microbiology and Molecular Biology Review, 64, 655-671). The effectiveness of antagonism based on the release of inhibitors into the environment is largely dependent on the conditions under which the experiment is conducted. Biedrzycka et al. (Biedrzycka E., Markiewicz LH, Bielecka M., Siwicki AK, 2007. Shaping the micro-ecosystem of the digestive tract, Controlling the development of the digestive system in newborn mammals, edited by Zabielski R., Wydawnictwo Rolne i Leśne, 5, 126-140) showed that the antagonistic activity of probiotic strains is not only the production of inhibitors, but also the prevention of colonization through the coaggregation of probiotic bacteria cells with pathogen cells and competition for the site of attachment to the host mucosa. Various mechanisms are involved in this process, e.g. electrostatic interactions, hydrophobic interactions, lipoteichoic acids (Gómez R., Geovanny D., Balcazar JL, Shen M, 2007. Probiotics as control agents in aquaculture, Journal of Ocean University of China, 6 , 76-79). These properties enable and help bacteria to maintain a significant advantage in the digestive tract of humans and animals. The dynamic and highly complex response of microorganisms is extremely important for intestinal epithelial cells and the host immune system (Shi HN, Walker A. Bacterial colonization and the development of intestinal defenses, 2004, Can. J Gastroenterol, 18, 493-500). It enables the maintenance of homeostasis and initiates the appropriate reactions of the organism against pathogens.

The digestive tract of humans and farm animals (poultry, cattle, pigs, horses, sheep, goats, etc.) may differ anatomically and functionally. However, a certain similarity of the intestinal microflora was found, both in terms of the amount and the presence of the same dominant groups of bacteria. Among the microflora colonizing the intestines of animals, there are many commensal bacteria, which are pathogenic to the animal organism and are the cause of zoonoses. The most common pathogenic bacteria of animal origin include: Salmonella, E. coli 0157: H7, Campylobacter, others such as Yersinia enterolitica, Listeria monocytogenes, Bacillus cereus, Staphylococcus aureus, Shigella sp. And Clostridium sp., Which may or may not be carried by food of animal origin. In a pioneering work entitled Probiotics: Intestinal inoculants for production animals provides a discussion of the available probiotics for the rearing of various animals, including chickens, pigs, and calves (Fox Veterinary Medicine, 08/1988). WO 89/05849 describes acid and bile tolerant Lactobacillus bacteria from the gastrointestinal tract of pigs which, according to the authors, can be used to ferment milk which can then be fed to piglets to prevent diarrhea. The publications of US 5 705 160, EP 353581, PL 179 838, PL 195 089, PL 214 583 describe the properties of many Lactobacillus plantarum strains having the properties of producing large amounts of bacteriostatic substances.

Infections with Salmonella bacteria are one of the most common human diseases transmitted through food, contaminated poultry products are one of the main sources of infection. Efforts to control Salmonella infections in domestic poultry are due, with few exceptions, to public health concerns and, to a lesser extent, the desire to significantly increase poultry production efficiency. Therefore, it is considered justified and very desirable to use natural additives in poultry nutrition, the effect of which will be to reduce the number of Salmonella bacteria in the content of the intestinal digestive tract.

It has been shown that the applied probiotic preparations exert a beneficial effect on the skin through their action on the "gut-brain-skin" axis. According to this assumption, an improper diet, poor in dietary fiber, stress, and antibiotic therapies cause excessive growth of potentially pathogenic and / or pathogenic microorganisms in the gut. As a result, the intestinal barrier is weakened and toxic substances that cause inflammation are absorbed into the bloodstream. Which in people predisposed to acne vulgaris, rosacea and allergies may lead to exacerbation of skin lesions. Therefore, it is justified to use probiotic bacteria with proven properties, in creams or ointments as the so-called a specific "shield" protecting against pathogens. This action is based on inhibiting the colonization of pathogens present on the skin by blocking their adherence with the simultaneous production of antibacterial substances. In addition, probiotic bacteria inhibit the immune response and, as a result, reduce inflammation in the skin. In addition, they break down sebum and other skin secretions, thus making it easier to absorb the nutrients contained in the cream.

The aim of the invention is to isolate a new strain of Lactobacillus plantarum with probiotic properties, and in particular showing bactericidal properties against potential and / or pathogens in humans and animals. The strain will have optimal effects on the digestive system and skin of humans and animals.

The essence of the invention is a new Lactobacillus plantarum AMT12 strain and a composition containing a new Lactobacillus plantarum AMT12 strain and carrier, distinguished by outstanding pro-biotic properties, with particular emphasis on antagonistic activity against bacteria belonging to the species Staphylococcus aureus, Salmonella enterica, Escherichia coli, which are pathogens of humans and animals. Used in the production of creams and ointments, parapharmaceutical, pharmaceutical and food preparations / products as well as food and water additives for humans and animals. The Lactobacillus plantarum AMT12 strain has been deposited in the Polish Collection of Microorganisms at the Institute of Immunology and Experimental Therapy of the Polish Academy of Sciences in Wrocław, deposit number B / 00107.

Strain identification

The Lactobacillus plantarum AMT12 strain comes from a plant environment. The strain was deposited in accordance with the provisions of the Budapest Treaty at the Polish Collection of Microorganisms (PCM) at the Institute of Immunology and Experimental Therapy of the Polish Academy of Sciences. The deposit was made on 03-02-2016 and marked with the number B / 00107.

Determination of the species affiliation of the Lactobacillus plantarum AMT12 strain In order to investigate the species affiliation of the Lactobacillus plantarum AMT12 strain, genotyping was performed by PCR using species-specific primers.

PCR genotyping

Identification of the isolated strains was performed using the DNA sequencing method. The secured material was used for DNA isolation.

DNA isolation

DNA isolation was performed according to the procedure: 1. 1 ml of bacterial culture was transferred to an Eppendorf tube, centrifuged (1 min, maximum rotation) and the supernatant was poured out. This operation was repeated 3 times. 2. 100 µl of LT lysis buffer and 10 µl of proteinase K and lysozyme were added to the tube with cells. 3. The tube was incubated at 50 ° C for 60 minutes (mixing by vortexing every 15 minutes). 4. After incubation, the tube was vortexed vigorously for 20 seconds. 5. The sample was centrifuged for 3 minutes (12,000 revolutions, Eppendorf centrifuge). 6. The supernatant was poured into a previously described DNA purification minicolumn. 7. Centrifuge for 1 minute (12k rpm, Eppendorf centrifuge). 8. 500 µΙ of A1 washing solution was added to the minicolumn. 9. Centrifuge for 1 minute (12k rpm, Eppendorf centrifuge). 10. The minicolumn was transferred to a new tube (2 ml) and 300 µl of washing solution A1 was added to the minicolumn. 11. Centrifugation for 3 minutes (12k revolutions, Eppendorf centrifuge). 12. The minicolumn was transferred to a new tube (1.5 ml) and 60 µL of deionized water was added to the bed at the bottom of the minicolumn. 13. Incubate the sample for 5 minutes at room temperature. 14. Centrifuge for 1 minute (12k revolutions, Eppendorf centrifuge). 15. The minicolumn was removed and the purified DNA in the tube was stored in the refrigerator. 16. DNA quality and quantity were analyzed by spectrophotometric method.

PCR and DNA sequencing

The isolated DNA was amplified by PCR

Components of the PCR mixture: 1. 10 x DNA polymerase buffer 6 μΙ 2. MgCb 25 mM, 2.4 μΙ 3. Free nucleotides 2 mM, 1.3 μΙ 4. primer R 20 pmol, 0.5 μΙ 5. primer F 20 pmol, 0.5 μΙ 6. Η2Ο15μΙ 7. DNA polymerase 2u / 1 μΙ, 0.15 μΙ

Starter R: 341: 5 '- CCTACGGGAGGCAGCAG -3' (Muyzer et al. 1993)

Starter F: 16SR: 5 '- TACCTTGTTACGACTTCACCCCA-3' (Rossau et al. 1991)

The obtained PCR products were subjected to a sequencing reaction which was carried out in a specialized laboratory of GENOMED (Warsaw, 12 Ponczowa Street). Elmer ABI 373 Automated DNA Sequencer (PE Applied Biosystems, FosterCity, CA, USA). As a result of sequencing, the DNA sequence of the 16S rRNA gene fragment of each of the analyzed strains was obtained.

GTCTGATGGAGCACGCCGCGTGAGTGAAGAAGGGTTTCGGCTCGTAAAACT

CTGTTGTTAAAGAAGAACATATCTGAGAGTAACTGTTCAGGTATTGACGGT

ATTTAACCAGAAAGCCACGGCTAACTACGTGCCAGCAGCCGCGGTAATACG

TAGG TGGCAAGCGTTGTCCGGATTTATTGGGCG TAAAGCGAGCGCAGGCGG

TTTTTTAAGTCTGATGTGAAAGCCTTCGGCTCAACCGAAGAAGTGCATCGG

AAACTGGGAAACTTGAGTGCAGAAGAGGACAGTGGAACTCCATGTGTAGC

GGTGAAATGCGTAGATATATGGAAGAACACCAGTGGCGAAGGCGGCTGTC

TGGTCTGTAACTGACGCTGAGGCTCGAAAGTATGGGTAGCAAACAGGATTA

GATACCCTGGTAGTCCATACCGTAAACGATGAATGCTAAGTGTTGGAGGGT

TTCCGCCCTTCAGTGCTGCAGCTAACGCATTAAGCATTCCGCCTGGGGAGT

ACGGCCGCAAGGCTGAAACTCAAAGGAATTGACGGGGGCCCGCACAAGCG

GTGGAGCATGTGGTTTAATTCGAAGCTACGCGAAGAACCTTACCAGGTCTT

GACATACTATGCAAATCTAAGAGATTAGACGTTCCCTTC

Each of the approximately 650 bp sequences obtained was identical. Comparative analysis with the DNA sequences deposited with the Gene Bank (NCBI) showed that the analyzed sequence is identical to the sequence of Lactobacillus plantarum.

The conducted studies with the genotypic method confirmed that the tested AMT12 strain belongs to the species Lactobacillus plantarum.

In vitro study of the antagonistic activity of Lactobacillus plantarum AMT12 against pathogens of the gastrointestinal tract and skin Strains of the species Salmonella enterica subsp. enterica serovar Enteritidis (strains: Salmonella enterica subsp. enterica serovar Enteritidis KOS 64, Salmonella enterica subsp. enterica serovar Enteritidis 65 / s / 10) and Escherichia coli (strains: Escherichia coli 0157: H7 - endocrine haemorrhagic strain, Escherleichia coli 1917 - a strain isolated from a commercial dietary supplement called Mutaflor), Staphylococcus aureus ATTC 33862. The pathogenic strains used in in vitro studies came from the National Salmonella Center in Gdynia (Salmonella enterica subsp. enterica serovar Enteritidis KOS 64), Veterinary Hygiene Department (Salmonella enterica subsp enterica serovar Enteritidis 65 / s / 10 - field strain, isolated from sick birds) and the Institute of Animal Reproduction and Food Research of the Polish Academy of Sciences in Olsztyn (Escherichia coli 0157: H7, Escherichia coli Nissle 1917, Staphylococcus aureus ATTC 33862).

The L. plantarum AMT12 strain came from the own collection of microorganisms of PROBIOS Sp. z o. o. The pathogenic strains were stored in the form of a lyophilisate at 4 ° C and immediately prior to testing, they were activated by two passages in a liquid tryptone-soy medium (TSB, Merck, cat. no. 1054590500). In order to determine the antibacterial capacity of L. plantarum AMT12 bacteria against selected pathogens belonging to the species Salmonella enterica subsp. enterica serovar Enteritidis, Escherichia coli and Staphylococcus aureus were cultivated together in a liquid tryptic-soy medium (TSB, Merck, cat. No. 1054590500) ensuring good growth of the inhibitory strain - Lactobacillus plantarum AMT12 and inhibited - Salmonella enterica subsp. enterica serovar Enteritidis KOS 64, Salmonella enterica subsp. enterica serovar Enteritidis 65 / s / 10, Escherichia coli O157: H7, Escherichia coli Nissle 1917, Staphylococcus aureus ATTC 33862.

The co-cultures (tested samples) were inoculated with a lyophilized strain of Lactobacillus plantarum AMT12 at the level of 109 colony forming units / ml (described further by the commonly accepted Polish abbreviation cfu / ml) and an active monoculture of the pathogenic strain of the species Salmonella enterica subsp. enterica serovar Enteritidis in the amount of 105 to 106 cfu / ml, Escherichia coli in the amount of 106 to 107 cfu / ml, Staphylococcus aureus in the amount of 106 cfu / ml. Control samples in the experiment were single strains of pathogenic bacteria (Salmonella enterica subsp. Enterica serovar Enteritidis KOS 64, Salmonella enterica subsp.enterica serovar Enteritidis 65 / s / 10, Escherichia coli O157: H7, Escherichia coli Nissle 1917, Staphylococcus aureus ATTC 33862 aureus) and Lactobacillus plantarum AMT12 monoculture using the inoculum level and liquid medium as in co-cultures.

Common and single cultures were prepared in four parallel tubes in triplicate. Incubation was carried out in aerobic conditions at 37 ° C for 0 hours (the so-called blank test determining the inoculum) for 24, 48 and 72 hours. After incubation, the number of viable Lactobacillus plantarum bacteria and pathogens in co-cultures and controls was determined by the plate method using appropriate agar media (Table No. 1). The material to be tested was diluted with 1% peptone water using the serial tenfold dilution method and plated on the bottom of a petri dish, then covered with liquefied agar medium at a temperature of about 45 ° C. Immediately after the medium solidified, the plates were turned upside down and incubated at 37 ° C for 24 or 48 hours aerobically or relatively anaerobically.

The culture conditions are given in Table 1.

After incubation, the colonies of bacteria in co-cultures were counted and compared with the number of bacteria in control (single) cultures.

Table 1

Cultivation conditions of the tested bacterial strains

The results of in vitro studies of the antagonistic activity of the Lactobacillus plantarum AMT12 strain against pathogenic factors of the gastrointestinal tract and skin are presented in Table 2.

Table 2

In vitro study of the antagonistic activity of Lactobacillus plantarum AMT12 against pathogens of the gastrointestinal tract and skin

* A! B - absent in 1 ml of culture In the conducted in vitro studies a total reduction in the number of Salmonella enterica subsp. enterica serovar Enteritidis KOS64, Salmonella enterica subsp. enteńca serovar Enteritidis 65 / s / IO, Escherichia coli O157: H7, Escherichia coli Nissle 1917 / Staphylococcus aureus ATTC 33862 after 24-hour incubation with Lactobacillus plantarum AMT12.

Determination of the multiplication capacity of Lactobacillus plantarum AMT12 in the presence of mycotoxins, deoxynivalenone and zearalenone. Liquid MRS medium (Merck, cat. Aldrich Cat # 32943-5MG) or Zearalenone (Sigma-Aldrich Cat # 32939-5MG) with a final assay concentration of 2 μg / ml each.

The cultivation was carried out in three parallel tubes under aerobic conditions at 37 ° C for 24 and 48 hours. Additionally, a control culture of the tested strain was carried out in parallel without the addition of the tested mycotoxins under aerobic conditions and optimal growth temperature, ie 37 ° C. The number of viable cells was determined immediately after inoculation and after 24 and 48 hours. The incubation of the bacteria in the petri dishes was carried out at 37 ° C for 48 hours under relatively anaerobic conditions.

The data are presented in Table 3

Table 3

Determination of the multiplication capacity of Lactobacillus plantarum AMT12 in the presence of mycotoxins, deoxynivalenone and zearalenone

Lactobacillus plantarum AMT12 showed the ability to grow in the presence of the tested mycotoxins, i.e. deoxynivalenone and zearalenone, in relation to the control culture of the AMT12 strain carried out in parallel (Table 3). On the first day of incubation, the number of L. plantarum AMT12 was at the level of 109 colony-forming units / ml (CFU / ml). At 48 hours of incubation, all tested samples showed a decrease in the number of AMT12 strain bacteria to the level of 108 cfu / ml.

Based on the conducted research, it can be assumed that the Lactobacillus plantarum AMT12 strain has unique growth and multiplication properties in the presence of one of the most dangerous mycotoxins, deoxynivalenone or zearalenone.

Determination of the growth capacity of Lactobacillus plantarum AMT12 in the presence of selected antibiotics most commonly used in animal therapy

Testing the growth capacity of Lactobacillus plantarum AMT12 in the presence of selected 20 antibiotics was performed using the co-culture method. For this purpose, the liquid medium M RS (Merck, catalog no. 1106610500) was inoculated with Lactobacillus plantarum AMT12 at a dose of 2.1 × 106 colony-forming units / ml and one of the 20 antibiotics at a concentration corresponding to the dose of antibiotic used in animals per kilogram of weight body (Table No. 4).

The cultivation was carried out in three parallel tubes under aerobic conditions at 37 ° C for 24 hours. Additionally, a control culture of the tested strain was carried out in parallel without the addition of the tested antibiotics. The number of viable cells was determined immediately after inoculation and after 24 hours.

Data on the growth capacity of Lactobacillus plantarum AMT12 in the presence of selected antibiotics used in animal therapy are presented in Table 4.

Table 4

Determination of the growth capacity of Lactobacillus plantarum AMT12 in the presence of selected antibiotics most commonly used in animal therapy

Lactobacillus plantarum AMT12 showed a unique broad spectrum of antibiotic resistance to the tested antibiotics. In the presence of 11 antibiotics (tiamulin hydrogen fumarate, enrofloxacin, colistin, tilmicosin, toltrazuril, amprolium, levamisole hydrochloride, flubendazole, neomecin, sodium sulphanopyrazine, lincomycin), out of 20 tested antibiotics, an excellent multiplication capacity was found the strain achieved in a control culture. However, in the presence of florfenicol, doxycillin, phenoxymethylpenicillin and tylosin, L. plantarum AMT12 abundance remained at the inoculum level. In the case of co-cultures with sulfmethoxazole, amoxicillin, lincomycin in combination with spectrinomycin, tylvalosin, the number of AMT12 bacteria decreased by 1 to 3 orders of magnitude compared to the control. Lactobacillus plantarum AMT12 showed no antibiotic resistance to only one two-component preparation, i.e. amoxicillin with the addition of clavulanic acid.

Determination of the multiplication capacity of Lactobacillus plantarum AMT12 at the following temperatures: 15 and 20 ° C. Liquid MRS medium (Merck, cat. No. 1106610500) was inoculated with the Lactobacillus plantarum AMT12 strain at a dose of about 107 colony-forming units / ml. The cultivation was carried out in three parallel tubes under aerobic conditions at temperatures of 15 and 20 ° C for 24, 48 and 72 hours. Additionally, a control culture of the tested strain was carried out in parallel under aerobic conditions and optimal growth temperature, ie 37 ° C. The number of viable cells was determined immediately after inoculation and after 24, 48 and 72 hours.

Table 5

Determination of the possibility of growth of the Lactobacillus plantarum AMT12 strain at 15 and 20 ° C

Lactobacillus plantarum AMT12 showed the ability to grow at 15 and 20 ° C. The tested Lactobacillus plantarum AMT12 strain was slightly slower to multiply on the first day of incubation at 15 ° C, in which it reached the number of 1.4 x 108 colony-forming units / ml (CFU / ml), in relation to the control culture carried out in parallel - 1.5 x 109 cfu / ml. At 48 and 72 hours of incubation, its number was 1.6 x 109 and 2.3 x 109 cfu / ml, respectively. However, in the control culture, on the following days of incubation, a decrease in the number of cells to the level of ~ 108 cfu / ml was found. On the other hand, the size of the L. plantarum AMT12 population at 20 ° C was comparable to the number of cells achieved by the AMT12 strain at the optimal growth temperature, ie 37 ° C.

After 24 and 48 hours of incubation at 20 ° C, its number was respectively: 2.0 χ 109, 2.9 x 109 cfu / ml and remained unchanged on the next day of incubation.

It was found that the Lactobacillus plantarum AMT12 strain has unique growth properties at low temperatures, i.e. below 16 ° C.

Survival determination of Lactobacillus plantarum AMT12 strain at low pH and in the presence of bile salts

The survival of the Lactobacillus plantarum AMT12 strain at low pH was determined by reducing the acidity of the Lactobacillus plantarum AMT12 culture in the stationary growth phase to a pH value of 3. However, in the case of determining the survival of the Lactobacillus plantarum AMT12 strain in the presence of bile salts, the pH values of the Lactobacillus plantarum AMT12 culture were initially increased. to a value of 6, then added bile salts in an amount of 3%

breeding. Determinations of the survival of the Lactobacillus plantarum AMT12 strain at low pH were performed before lowering the pH of the culture (control test) just after lowering the pH of the culture to a value equal to 3, the so-called 0 minutes followed by 40 and 180 minutes incubation at 37 ° C in anaerobic conditions. The survival of Lactobacillus plantarum AMT12 in the presence of bile salt was performed before the addition of bile salt (control test), just after the addition of bile salt, the so-called 0 minutes and after 1.3 and 6 hours of incubation at 37 ° C in anaerobic conditions. Viable Lactobacillus plantarum AMT12 cells were determined in colony forming units (CFU / ml) using the pour plate technique. The survival of the Lactobacillus plantarum AMT12 strain was expressed as a percentage of the number of Lactobacillus plantarum AMT12 cells after 180 minutes for the survival determination at pH 3 and after 6 hours for the determination of the number of Lactobacillus plantarum AMT12 in the presence of bile salt, compared to the number of the Lactobacillus strain, respectively plantarum AMT 12 under control.

Table 6

Survival of Lactobacillus plantarum AMT12 at pH = 3

Table 7

Survival of Lactobacillus plantarum AMT12 strain in the presence of 3% bile salts

The tested Lactobacillus plantarum AMT12 strain showed 100% survival at low pH = 3 and 89% survival in the presence of bile salt concentration.

The results showed high resistance of the Lactobacillus plantarum AMT12 strain to low pH and to bile salts, which proves that this strain is adapted to the conditions in the digestive tract.

The following examples illustrate the invention.

Example 1

The probiotic composition, which is an additive to poultry feed or water, is a product in the form of freeze-dried bacteria and a binder, and as a bacterial strain it contains the Lactobacillus plantarum AMT12 PCM B / 00107 strain in the amount of 2.5 x 106 colony forming units - cfu / ml.

Example 2

The probiotic composition, which is an additive to fodder or water for aquaculture, is a product in the form of freeze-dried bacteria and a binder, and contains as a bacterial strain Lactobacillus plantarum AMT 12 PCM B / 00107 in the amount of 2.0 x 107 colony forming units - cfu / ml.

Claims (2)

Patent claims 1. Lactobacillus plantarum AMT12 strain, PCM B / 00107. 2. Composition for the production of creams and ointments, parapharmaceutical, pharmaceutical, food preparations / products as well as food and water additives for humans and animals, consisting of a new strain of carrier bacteria and a filler, characterized in that the bacterial strain comprises Lactobacillus plantarum AMT12 deposited in PCM B / 00107 in an amount of 103 to 1013 colony forming units / g.