WO2011159178A1 - New strain of lactobacillus plantarum s, the use of the strain of lactobacillus plantarum s and the preparation for roughages ensiling - Google Patents
New strain of lactobacillus plantarum s, the use of the strain of lactobacillus plantarum s and the preparation for roughages ensiling Download PDFInfo
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- WO2011159178A1 WO2011159178A1 PCT/PL2011/000059 PL2011000059W WO2011159178A1 WO 2011159178 A1 WO2011159178 A1 WO 2011159178A1 PL 2011000059 W PL2011000059 W PL 2011000059W WO 2011159178 A1 WO2011159178 A1 WO 2011159178A1
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
- C12N1/205—Bacterial isolates
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
- A23K10/16—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions
- A23K10/18—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions of live microorganisms
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/30—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K30/00—Processes specially adapted for preservation of materials in order to produce animal feeding-stuffs
- A23K30/10—Processes specially adapted for preservation of materials in order to produce animal feeding-stuffs of green fodder
- A23K30/15—Processes specially adapted for preservation of materials in order to produce animal feeding-stuffs of green fodder using chemicals or microorganisms for ensilaging
- A23K30/18—Processes specially adapted for preservation of materials in order to produce animal feeding-stuffs of green fodder using chemicals or microorganisms for ensilaging using microorganisms or enzymes
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/225—Lactobacillus
- C12R2001/25—Lactobacillus plantarum
Definitions
- the subject of the invention is a new strain of Lactobacillus plantarum S bacteria, its use and the preparation for ensiling of contaminated roughages.
- Ochratoxin A is synthesized by moulds from Aspergillus sp. and Penicillium sp., which are often developed on agricultural products. OTA has been classified by the International Agency for Research on Cancer as carcinogenic factor of category 2B. The monitoring studies, as conducted in different years and concerning OTA presence in feedstuffs and food, indicate the existence of serious hazards for animals and humans, especially during hot and wet years, being favorable for growth of moulds.
- the purpose of the invention was to select a strain of lactic acid bacteria, characterized by the ability of growing in the environment contaminated with ochratoxin A and reducing OTA level by at least 80% in order to apply it in ensiling of the contaminated roughages.
- the strain according to the invention was isolated from the spontaneous fermentating maize plants. It was classified to Lactobacillus plantarum species on the basis of studying morphological and physiological-biochemical properties with API 50 CH kit (Biomerieux, France) and selected in order to obtain the ability of developing and synthesizing organic acids in growth media, containing ochratoxin A in concentration of 50 - 100 ppb.
- the object of the invention is the new bacterial strain, which was specified as Lactobacillus plantarum S and deposited in the IAFB Collection of Industrial Microorganisms Institute of Agricultural and Food Biotechnology in Warsaw under the number KKP 2021 p or variant thereof.
- the second object of the invention is the composition comprising an effective amount of Lactobacillus plantarum S deposited under the number KKP 2021 p or variant thereof.
- Preferred composition further comprises at least one carrier and/or at least one emulsifier and/or at least bio-stimulator, preferably is in dry form
- the third object of the invention includes granular or powder preparation for ensiling of the roughages which contains dried-on-the carrier biomass of Lactobacillus plantarum S KKP 2021 p with bacterial number equal to 9 - 10 log CFU/g of the preparation; the composition of the carriers contains extract from aloes which constitutes 0.2 - 2.0 % of dry matter of the preparation.
- Another object of the invention is a method of ensiling of roughages, which comprises the step of ensiling of roughages with use of new strain of Lactobacillus plantarum S KKP 2021 p according to the invention and/or the composition according to the invention and/or preparation according to the invention.
- Another object of the invention is the use of the new strain of Lactobacillus plantarum S KKP 2021 p or variant thereof for ensiling of roughages, intended for decontamination of roughages, contaminated with ochratoxin A and/or moulds and/or with pathogenic bacteria selected from genus Salmonella and/or genus Listeria and/or coliform bacteria and/or bacteria of the species Escherichia coli and/or Clostridium perfringens.
- Another object of the invention is the use of new strain of Lactobacillus plantarum S KKP 2021 p according to the invention and/or the composition according to the invention and/or preparation according to the invention for ensiling of roughages.
- ensiling of roughages is carried out to decontaminate or/and restrain from contamination and/or prevent contamination of roughages, contaminated with ochratoxin A and/or moulds and/or with pathogenic bacteria selected from genus Salmonella and/or genus Listeria and/or coliform bacteria and/or bacteria of the species Escherichia coli and/or Clostridium perfringens.
- mutant strain or strains obtained by rising the deposited strain as starter material wherein the mutant strain(s) retain or further improve the ability of growing and activity of parent strain i.e. ability to grow in the environment contaminated with ochratoxin A and reducing OTA level and/or reducing moulds and/or reducing pathogenic bacteria growth.
- Fig.l Shows changes in ochratoxin A content in medium during cultivation of Lactobacillus plantarum S KKP 2021 p.
- Fig.2. Shows survival of pathogenic bacteria during their incubation with the strain Lactobacillus plantarum S KKP 2021 p.
- the new strain according to the invention was isolated from the spontaneous fermentating maize plants. It was classified to Lactobacillus plantarum and deposited in the International Collection of Industrial Microorganism Cultures of the Institute of Agricultural and Food Biotechnology under the number KKP 2021 p.
- Lactobacillus plantarum S KKP 2021 p strain is characterized by the following features:
- Bacterial cells have a form of short rods occurring singly, in pairs or in chains of 0.9 - 1.2 um. They do not produce spores and are Gram+.
- the new strain is relatively heterofermentative, microaerophilic, does not produce catalase, does not reduce nitrates to nitrites, and is capable of fermenting the following sugars: saccharose, galactose, glucose, fructose, arabinose, maltose, mannose, esculin, lactose, cellobiose, gentiobiose, mellibiose, rafinose, ribose, xylose, sorbitol and trehalose.
- the sequence of 16S rDNA segment of Lactobacillus plantarum S KKP 2021 p is given on SEQ ID NO: l .
- the new strain is characterized by the ability of developing and synthesizing lactic acid in medium, containing ochratoxin A and of removing it from the environment in at least 80% as well as inhibiting growth of moulds. Moreover, it was revealed that the new strain showed, simultaneously, a very high antibacterial activity against pathogenic bacteria from Salmonella and Listeria genus, Escherichia coli and Clostridium perfringens, which are often present in agricultural products, cultivated on organically fertilized land, especially with the use of liquid organic manure.
- Antibacterial activity of the strain Lactobacillus plantarum S KKP 2021 p in relation to pathogenic strains Salmonella sp. and Escherichia coli, isolated from digestive tract of sick animals was determined by Pilet method (Pilet et al., 1995, J. Food Prot., 58, 256 - 262). After 24 hours of cultivation of the examined strain, 10 ⁇ of supernatant were introduced onto the surface of agar medium with the diameter of 10 cm, containing Salmonella sp. or Escherichia coli. After the successive 24 hours, the diameter of bacterial growth inhibition zone was measured. In the experimental conditions metabolites, produced by the strain Lactobacillus plantarum S KKP 2021 p, inhibited the growth of tested pathogenic bacteria in the range of 90 - 100%.
- Bacterial cultures cultivated for 24 h were employed as the experimental material.
- the cultures of Salmonella sp. and Escherichia coli, with the number of bacteria 6.5 and 6.0 log CFU/ml, respectively, were combined with the culture of Lactobacillus plantarum S KKP 2021 p bacteria with the number of bacteria 9.0 log CFU/ml in v/v ratio 1 :1.
- the new strain Lactobacillus plantarum S KKP 2021 p inhibited completely Salmonella sp. growth and after seven days of incubation - development of Escherichia coli (see Fig.2).
- Cultivation of biomass of Lactobacillus plantarum S KKP 2021 p was conducted in fermentors with volume of 150 1, at temperature of 30°C, at pH 5.7, regulated by ammonia water.
- the content of the components of culture medium in g/1 water was as follows: glucose - 20, yeast extract - 10, maize soak - 5, ammonium biphosphate - 0.2, ammonium sulphate - 0.35 and manganese sulphate - 0.01.
- Bacteria were cultivated for 24 h. Bacteria biomass with ca.
- 30% DM obtained after separation of culture medium at rotations of separator equal to 14000/min., was mixed with the following carriers: mix of starter sugars: saccharose, glucose, lactose, soluble potato starch, emulsifier (lecithin) and bio-stimulator (aloes extract). Then, it was dried in fluidal drier at temperature up to 40°C.
- the dried bacterial preparation contained ca. 94% DM, including: 90% sugars; 0.2% of dried aloes extract and 3.8% of dried bacterial biomass and was characterized by the number of bacteria equal to 9.0 log CFU/g (colonies- forming units).
- silages In the experimental agricultural farm, two types of silages were produced: from meadow sward and maize plants with humidity of ca. 40%, contaminated with ochratoxin A, moulds and pathogenic bacteria.
- the preparation, according to the invention obtained in example I
- the preparation, according to the invention was applied in a form of water spray.
- the samples of silages without and with the addition of the preparation were collected.
- the influence of Lactobacillus plantarum S KKP 2021 p on reduction of ochratoxin A, number of moulds and pathogenic bacteria in the ensiled feedstuffs were examined.
- the number of moulds was lowered by 77% in the silages produced from meadow sward and by 80 % in the silages from maize plants as compared to the silages obtained without the addition of the preparation Lactobacillus plantarum S KKP 2021 p.
- the contaminated with ochratoxin A and pathogenic microorganisms feeds after the process of lactic fermentation provided by bacteria of the new strain Lactobacillus plantarum S KKP 2021 p, may be employed as completely safe products in feeding of breeding animals.
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Abstract
The invention relates to the new, genetically non-modified bacterial strain Lactobacillus plantarum S, which has been deposited in the IAFB Collection of Industrial Microorganisms Institute of Agricultural and Food Biotechnology in Warsaw under the number KKP 2021 p or variant thereof. The invention relates to use of the strain Lactobacillus plantarum S KKP 2021 p or variant thereof or composition comprising thereof for ensiling of roughages, with the intention to decontaminate the feeds, contaminated with ochratoxin A, moulds and pathogenic bacteria. The feeds contaminated with moulds, ochratoxin A and pathogenic microorganisms, after the process of lactic fermentation with the participation of bacteria of the new strain Lactobacillus plantarum S, may be employed in nutrition of breeding animals as being completely safe products.
Description
New strain of Lactobacillus plantarum S, the use of the strain of Lactobacillus plantar uni S and the preparation for roughages ensiling
DESCRIPTION TECHNICAL FIELD
The subject of the invention is a new strain of Lactobacillus plantarum S bacteria, its use and the preparation for ensiling of contaminated roughages.
BACKGROUND ART
Ochratoxin A (OTA) is synthesized by moulds from Aspergillus sp. and Penicillium sp., which are often developed on agricultural products. OTA has been classified by the International Agency for Research on Cancer as carcinogenic factor of category 2B. The monitoring studies, as conducted in different years and concerning OTA presence in feedstuffs and food, indicate the existence of serious hazards for animals and humans, especially during hot and wet years, being favorable for growth of moulds.
From the patent specifications WO 91/13555 and WO 92/05706 methods of binding the mycotoxins in feedstuffs and digestive tract- of animals by mineral compounds are known. Use of the mentioned adsorbents bind also water as well as dissolved in water and important for organisms nutrients and vitamins. It is technically impossible to employ the currently known chemical and physical methods of decontamination of the feed, contaminated with OTA directly in agricultural farms, or they are not admitted to application in the EU countries. For that reason biological methods for fodder decontamination, which are based on the application of the selected strains of bacteria and/or yeasts, are demanded.
In the patent specification of US 2004/0208956 Al following strains of microorganisms are mentioned: Shingomonas, Stenotrophomonas, Ochrabactrum, Ralstonia and/or Eubacterium and yeasts: Trichosporon, Cryptococcus and Rhodotorula. These microorganisms are capable of ochratoxins detoxification by enzymatic cleaving of phenylalanine.
Naturally living in the environment and not being genetically modified new strains of lactic acid bacteria might be employed for the improvement of safety of ensiled feeds and in consequence of food. This has become the subject of constantly undertaken studies. The discussed abilities are strictly connected with the specified strains of lactic acid bacteria and cannot be identified with any species or genus of lactic acid bacteria. The mentioned group of microorganisms contains innumerous bacterial strains which are characterized - to a different
extent - by the ability to lower OTA content in the model environment. On the other hand, there is a lack of the results of their practical application for decontamination of OTA-infected feeds, especially in the manufacturing conditions of agricultural farms.
It is impossible to protect the agricultural products from contamination with toxin- producing moulds. For that reason simple biological methods for eliminating or reducing OTA content by the selected bacterial strains from Lactobacillus sp. with GRAS status are being searched. These bacteria might be employed as starter cultures in the process of feed ensiling. DISCLOSURE OF INVENTION
The purpose of the invention was to select a strain of lactic acid bacteria, characterized by the ability of growing in the environment contaminated with ochratoxin A and reducing OTA level by at least 80% in order to apply it in ensiling of the contaminated roughages.
The strain according to the invention, was isolated from the spontaneous fermentating maize plants. It was classified to Lactobacillus plantarum species on the basis of studying morphological and physiological-biochemical properties with API 50 CH kit (Biomerieux, France) and selected in order to obtain the ability of developing and synthesizing organic acids in growth media, containing ochratoxin A in concentration of 50 - 100 ppb.
The object of the invention is the new bacterial strain, which was specified as Lactobacillus plantarum S and deposited in the IAFB Collection of Industrial Microorganisms Institute of Agricultural and Food Biotechnology in Warsaw under the number KKP 2021 p or variant thereof.
The second object of the invention is the composition comprising an effective amount of Lactobacillus plantarum S deposited under the number KKP 2021 p or variant thereof. Preferred composition further comprises at least one carrier and/or at least one emulsifier and/or at least bio-stimulator, preferably is in dry form
The third object of the invention includes granular or powder preparation for ensiling of the roughages which contains dried-on-the carrier biomass of Lactobacillus plantarum S KKP 2021 p with bacterial number equal to 9 - 10 log CFU/g of the preparation; the composition of the carriers contains extract from aloes which constitutes 0.2 - 2.0 % of dry matter of the preparation.
Another object of the invention is a method of ensiling of roughages, which comprises the step of ensiling of roughages with use of new strain of Lactobacillus plantarum S KKP
2021 p according to the invention and/or the composition according to the invention and/or preparation according to the invention.
Another object of the invention is the use of the new strain of Lactobacillus plantarum S KKP 2021 p or variant thereof for ensiling of roughages, intended for decontamination of roughages, contaminated with ochratoxin A and/or moulds and/or with pathogenic bacteria selected from genus Salmonella and/or genus Listeria and/or coliform bacteria and/or bacteria of the species Escherichia coli and/or Clostridium perfringens.
Another object of the invention is the use of new strain of Lactobacillus plantarum S KKP 2021 p according to the invention and/or the composition according to the invention and/or preparation according to the invention for ensiling of roughages. Preferably ensiling of roughages is carried out to decontaminate or/and restrain from contamination and/or prevent contamination of roughages, contaminated with ochratoxin A and/or moulds and/or with pathogenic bacteria selected from genus Salmonella and/or genus Listeria and/or coliform bacteria and/or bacteria of the species Escherichia coli and/or Clostridium perfringens.
By the term variant of new strain Lactobacillus plantarum S KKP 2021 p it should be understood the mutant strain or strains obtained by rising the deposited strain as starter material, wherein the mutant strain(s) retain or further improve the ability of growing and activity of parent strain i.e. ability to grow in the environment contaminated with ochratoxin A and reducing OTA level and/or reducing moulds and/or reducing pathogenic bacteria growth.
BRIEF DESCRIPTION OF DRAWINGS
For a better understanding of the invention it will be described by the way of example only with reference to the accompanying drawings in which:
Fig.l. Shows changes in ochratoxin A content in medium during cultivation of Lactobacillus plantarum S KKP 2021 p.
Fig.2. Shows survival of pathogenic bacteria during their incubation with the strain Lactobacillus plantarum S KKP 2021 p.
DESCRIPTION OF EMBODIMENTS
The new strain according to the invention, was isolated from the spontaneous fermentating maize plants. It was classified to Lactobacillus plantarum and deposited in the
International Collection of Industrial Microorganism Cultures of the Institute of Agricultural and Food Biotechnology under the number KKP 2021 p.
Lactobacillus plantarum S KKP 2021 p strain is characterized by the following features:
• morphological - it creates colonies of the white to creamy colour, with a shape of discs of the size of 0.5 - 0.7 mm. Bacterial cells have a form of short rods occurring singly, in pairs or in chains of 0.9 - 1.2 um. They do not produce spores and are Gram+.
• physiological and biochemical - the optimal temperature of growth is 30°C. The new strain is relatively heterofermentative, microaerophilic, does not produce catalase, does not reduce nitrates to nitrites, and is capable of fermenting the following sugars: saccharose, galactose, glucose, fructose, arabinose, maltose, mannose, esculin, lactose, cellobiose, gentiobiose, mellibiose, rafinose, ribose, xylose, sorbitol and trehalose. Genetic analysis of the sequence of 16S rDNA segments and their comparison with sequences of bacterial 16S rDNA segment, deposited in the GenBank base, confirmed in 98% the taxonomic affiliation of the examined strain to Lactobacillus plantarum species, specified by biochemical properties. Genetic analysis involved a technique of molecular fingerprinting - RAPD-PCR based on randomly amplified polymorphic regions of genomic DNA. PCR reaction was carried out using the following primer: 5' GAGGGTGGCGGTTCT 3' and amplification conditions according to Andrighetto et. al. 2001 (Lett. Appl. Microbiol. 33, 26 - 30).
The sequence of 16S rDNA segment of Lactobacillus plantarum S KKP 2021 p is given on SEQ ID NO: l .
The new strain is characterized by the ability of developing and synthesizing lactic acid in medium, containing ochratoxin A and of removing it from the environment in at least 80% as well as inhibiting growth of moulds. Moreover, it was revealed that the new strain showed, simultaneously, a very high antibacterial activity against pathogenic bacteria from Salmonella and Listeria genus, Escherichia coli and Clostridium perfringens, which are often present in agricultural products, cultivated on organically fertilized land, especially with the use of liquid organic manure.
Cultivation of the new strain Lactobacillus plantarum S KKP 2021 p under model conditions in MRS medium at temperature of 30°C for 72 h resulted in reducing the content of ochratoxin A by 87.2% in relation to its initial level (see Fig. I).
Antibacterial activity of the strain Lactobacillus plantarum S KKP 2021 p in relation to pathogenic strains: Salmonella sp. and Escherichia coli, isolated from digestive tract of
sick animals was determined by Pilet method (Pilet et al., 1995, J. Food Prot., 58, 256 - 262). After 24 hours of cultivation of the examined strain, 10 μΐ of supernatant were introduced onto the surface of agar medium with the diameter of 10 cm, containing Salmonella sp. or Escherichia coli. After the successive 24 hours, the diameter of bacterial growth inhibition zone was measured. In the experimental conditions metabolites, produced by the strain Lactobacillus plantarum S KKP 2021 p, inhibited the growth of tested pathogenic bacteria in the range of 90 - 100%.
During 7 days of incubation, the survivability of the cultures of Salmonella sp. and Escherichia coli bacteria were evaluated.
Bacterial cultures cultivated for 24 h were employed as the experimental material. The cultures of Salmonella sp. and Escherichia coli, with the number of bacteria 6.5 and 6.0 log CFU/ml, respectively, were combined with the culture of Lactobacillus plantarum S KKP 2021 p bacteria with the number of bacteria 9.0 log CFU/ml in v/v ratio 1 :1. After six days of incubation, the new strain Lactobacillus plantarum S KKP 2021 p inhibited completely Salmonella sp. growth and after seven days of incubation - development of Escherichia coli (see Fig.2).
The invention is illustrated by the following examples. EXAMPLE 1
Cultivation of biomass of Lactobacillus plantarum S KKP 2021 p was conducted in fermentors with volume of 150 1, at temperature of 30°C, at pH 5.7, regulated by ammonia water. The content of the components of culture medium in g/1 water was as follows: glucose - 20, yeast extract - 10, maize soak - 5, ammonium biphosphate - 0.2, ammonium sulphate - 0.35 and manganese sulphate - 0.01. Bacteria were cultivated for 24 h. Bacteria biomass with ca. 30% DM, obtained after separation of culture medium at rotations of separator equal to 14000/min., was mixed with the following carriers: mix of starter sugars: saccharose, glucose, lactose, soluble potato starch, emulsifier (lecithin) and bio-stimulator (aloes extract). Then, it was dried in fluidal drier at temperature up to 40°C. The dried bacterial preparation contained ca. 94% DM, including: 90% sugars; 0.2% of dried aloes extract and 3.8% of dried bacterial biomass and was characterized by the number of bacteria equal to 9.0 log CFU/g (colonies- forming units).
EXAMPLE II
In the experimental agricultural farm, two types of silages were produced: from meadow sward and maize plants with humidity of ca. 40%, contaminated with ochratoxin A, moulds and pathogenic bacteria. To obtain the experimental silages, the preparation, according to the invention (obtained in example I), in the rate of 10 g/t of the cut green forage, was applied in a form of water spray. After six weeks and completion of ensiling process, the samples of silages without and with the addition of the preparation were collected. The influence of Lactobacillus plantarum S KKP 2021 p on reduction of ochratoxin A, number of moulds and pathogenic bacteria in the ensiled feedstuffs were examined. In the experimental silages with the addition of the preparation, the content of ochratoxin A in maize plants was lowered from 30.6 ppb to 4.5 ppb (85.3%); and in meadow sward - from 25.5 ppb to the level of 3.2 ppb (87.4%). In the silages, produced without the addition of the preparation, the content of ochratoxin A was lowered by 6.7 - 7.5%, depending on the initial concentration and type of the ensiled plants (tab.l)
Tab.l
Effect of the preparation on reduction of the content of ochratoxin A during the process of the roughages ensiling
Type of feedstuff and/or silage with DM The remaining
Content of ochratoxin A, ppb
ca. 40% content, %
Meadow sward - green forage 25.5 100.0
Meadow sward - silage A 23.8 93.3
Meadow sward - silage B 3.2 12.6
Maize plants - green forage 30.6 100.0
Maize plants - silage A 28.3 92.5
Maize plants - silage B 4.5 14.7
A - silages without the addition of the preparation
B - silages with the addition of the preparation
At the same time, microbiological purity of the silages produced without and with the addition of the preparation was examined; their microbiological contamination with bacteria from genus: Salmonella and Listeria, species: Escherichia coli and Clostridium perfringens as well as moulds, was determined. The results concerning the effect of the preparation on the improvement of the state of hygiene of the silages is given in tab.2.
Tab.2.
Microbiological contamination of ensiled roughages without and with the addition of the preparation
Number of pathogenic bacteria and moulds in silages, log CFU/g
Type of silage
produced from: Salmonella Listeria Escherichia Coliform Clostridium
Moulds sp. s/λ coli bacteria perfringens
Meadow sward-
2.60 3.88 3.30 2.50 3.70 5.60 A
Meadow sward- no presence 1.44 no presence no presence 1.00 1.30 B
Maize plants-A 2.30 3.60 2.70 3.30 2.30 5.20
Maize plants-B no presence 1.28 no presence 1.00 0.50 1.00
A - silages without the addition of the preparation
B - silages with the addition of the preparation
In the meadow sward silages produced with the addition of the preparation, pathogenic bacteria from genus Salmonella and the species Escherichia coli as well as coliform bacteria were eliminated in 100%. In the silages produced from maize plants with the addition of the preparation, the presence of Salmonella sp. and Escherichia coli were not found and coliform bacteria constituted only 30% of their number contained in the silage expressed in log CFU/g, produced without the preparation. The number of bacteria Clostridium perfringens in the silages with the addition of the preparation was lowered by 73 and 78%, respectively, and the number of Listeria sp. was lowered by 63 and 64 %, respectively. On the other hand, the number of moulds was lowered by 77% in the silages produced from meadow sward and by 80 % in the silages from maize plants as compared to the silages obtained without the addition of the preparation Lactobacillus plantarum S KKP 2021 p.
The contaminated with ochratoxin A and pathogenic microorganisms feeds, after the process of lactic fermentation provided by bacteria of the new strain Lactobacillus plantarum S KKP 2021 p, may be employed as completely safe products in feeding of breeding animals.
Claims
1. The new strain of Lactobacillus plantarum S, deposited in the IAFB Collection of Industrial Microorganisms Institute of Agricultural and Food Biotechnology in Warsaw under the number KKP 2021 p or variant thereof.
2. A composition of preparation comprising an effective amount of Lactobacillus plantarum S deposited under the number KKP 2021 p or variant thereof.
3. The composition according to claim 2, which further comprises at least one carrier and/or at least one emulsifier and/or at least one bio-stimulator.
4. The composition according to claim 3, wherein the bacterial biomass comprising Lactobacillus plantarum S KKP 2021 p or variant thereof is in dry form.
5. The preparation for ensiling roughages, in a granular form or powder, consisting of dried bacterial biomass and carriers, characterized in that it comprises the strain Lactobacillus plantarum S KKP 2021 p with bacterial number of 9 - 10 log CFU/g and the composition of the carriers contain aloes extract which constitutes 0.2 - 2.0 % of dry matter of the preparation.
6. A method of ensiling of roughages, wherein it comprises the step of ensiling of roughages with use of new strain of Lactobacillus plantarum S according to claim 1 and/or the composition according to claims 2-4 and/or preparation according to claim 5.
7. The use of the new strain of Lactobacillus plantarum S KKP 2021 p or variant thereof for ensiling of roughages, intended for decontamination of roughages, contaminated with ochratoxin A and/or moulds and/or with pathogenic bacteria selected from genus Salmonella and/or genus Listeria and/or coliform bacteria and/or bacteria of the species Escherichia coli and/or Clostridium perfringens.
8. The use of new strain of Lactobacillus plantarum S according to claim 1 and/or the composition according to claims 2-4 and/or preparation according to claim 5 for ensiling of roughages.
9. The use according to claim 8, wherein ensiling of roughages is carried out to decontaminate or/and restrain from contamination and/or prevent contamination of roughages, . contaminated with ochratoxin A and/or moulds and/or with pathogenic bacteria selected from genus Salmonella and/or genus Listeria and/or coliform bacteria and/or bacteria of the species Escherichia coli and/or Clostridium perfringens.
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EP11731146.4A EP2582851B8 (en) | 2010-06-16 | 2011-06-14 | New strain of lactobacillus plantarum s, the use of the strain of lactobacillus plantarum s and the preparation for roughages ensiling |
US13/684,983 US8697423B2 (en) | 2010-06-16 | 2012-11-26 | Strain of Lactobacillus plantarum S, the use of the strain of Lactobacillus plantarum S and the preparation for roughages ensiling |
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PL391534A PL214583B1 (en) | 2010-06-16 | 2010-06-16 | A new strain of Lactobacillus plantarum S, the use of the strain of Lactobacillus plantarum S and preparation for ensiling bulky feed |
PLPL391534 | 2010-06-16 |
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US13/684,983 Continuation-In-Part US8697423B2 (en) | 2010-06-16 | 2012-11-26 | Strain of Lactobacillus plantarum S, the use of the strain of Lactobacillus plantarum S and the preparation for roughages ensiling |
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US (1) | US8697423B2 (en) |
EP (1) | EP2582851B8 (en) |
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Cited By (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20140178528A1 (en) * | 2012-11-22 | 2014-06-26 | Instytut Biotechnologii Przemyslu Rolno-Spozywczego | Strain of lactobacillus buchneri a, composition, a multi-component preparation for starch-rich plant preservation, their use and a method for plant preservation |
WO2015014372A1 (en) * | 2013-07-30 | 2015-02-05 | OÜ Tervisliku Piima Biotehnoloogiate Arenduskeskus | Isolated microorganism strain lactobacillus plantarum tak 59 ncimb42150 and its use |
CN105861399A (en) * | 2016-06-17 | 2016-08-17 | 浙江大学 | Lactobacillus plantarum for preventing necrotic enteritis of chickens and application thereof |
CN112143670A (en) * | 2020-09-08 | 2020-12-29 | 中国农业科学院农产品加工研究所 | Lactobacillus brevis 8-2B and application thereof |
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KR101470995B1 (en) * | 2013-08-30 | 2014-12-09 | 주식회사 바이오리쏘스 | Propionic acid producing microorganism and roughage comprising the same |
PL241510B1 (en) | 2015-11-06 | 2022-10-17 | Probios Spolka Z Ograniczona Odpowiedzialnoscia | New Lactobacillus plantarum AMT14 strain and the composition containing the Lactobacillus plantarum AMT14 strain |
US10166262B2 (en) | 2017-04-20 | 2019-01-01 | Ewelina Sosnowska-Turek | Strain of bacteria and composition comprising the same |
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Citations (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4842871A (en) * | 1985-08-01 | 1989-06-27 | Pioneer Hi-Bred International, Inc. | Method and inoculant for preserving agricultural products for animal feed |
WO1991013555A1 (en) | 1990-03-07 | 1991-09-19 | Engelhard Corporation | Animal feed additive and method for inactivating mycotoxins present in animal feeds |
WO1992005706A1 (en) | 1990-10-01 | 1992-04-16 | Engelhard Corporation | Animal feed containing montmorillonite clay as additive |
EP1386543A1 (en) * | 2001-12-20 | 2004-02-04 | Medipharm AB | New bacterial strain and the use thereof |
US20040208956A1 (en) | 2001-12-20 | 2004-10-21 | Gerd Schatzmayr | Microorganism for biological detoxification of mycotoxins, namely ochratoxins and/or zearalenons, as well as method and use thereof |
WO2010017568A1 (en) * | 2008-07-21 | 2010-02-18 | Erber Aktiengesellschaft | Method for treating food silage for ruminants and food silage additive |
-
2010
- 2010-06-16 PL PL391534A patent/PL214583B1/en unknown
-
2011
- 2011-06-14 EP EP11731146.4A patent/EP2582851B8/en not_active Not-in-force
- 2011-06-14 WO PCT/PL2011/000059 patent/WO2011159178A1/en active Application Filing
-
2012
- 2012-11-26 US US13/684,983 patent/US8697423B2/en not_active Expired - Fee Related
Patent Citations (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4842871A (en) * | 1985-08-01 | 1989-06-27 | Pioneer Hi-Bred International, Inc. | Method and inoculant for preserving agricultural products for animal feed |
WO1991013555A1 (en) | 1990-03-07 | 1991-09-19 | Engelhard Corporation | Animal feed additive and method for inactivating mycotoxins present in animal feeds |
WO1992005706A1 (en) | 1990-10-01 | 1992-04-16 | Engelhard Corporation | Animal feed containing montmorillonite clay as additive |
EP1386543A1 (en) * | 2001-12-20 | 2004-02-04 | Medipharm AB | New bacterial strain and the use thereof |
US20040208956A1 (en) | 2001-12-20 | 2004-10-21 | Gerd Schatzmayr | Microorganism for biological detoxification of mycotoxins, namely ochratoxins and/or zearalenons, as well as method and use thereof |
WO2010017568A1 (en) * | 2008-07-21 | 2010-02-18 | Erber Aktiengesellschaft | Method for treating food silage for ruminants and food silage additive |
Non-Patent Citations (4)
Title |
---|
ANDRIGHETTO, LETT. APPL. MICROBIOL., vol. 33, 2001, pages 26 - 30 |
MALGORZATA PIOTROWSKA: "The Elimination of Ochratoxin A by Lactic Acid Bacteria Strains", POLISH JOURNAL OF MICROBIOLOGY, vol. 54, no. 4, 2005 - 2005, Poland, pages 279 - 286, XP055007437, Retrieved from the Internet <URL:http://www.pjm.microbiology.pl/archive/vol5442005279.pdf> [retrieved on 20110916] * |
PILET ET AL., J. FOOD PROT., vol. 58, 1995, pages 256 - 262 |
SUTERSKA: "EFFECT OF THE SELECTED STRAINS FROM LACTOBACILLUS GENUS ON THE LIMITATION OF MOULD AND OCHRATOXIN A CONTAMINATION OF SILAGES FROM MEADOW SWARD", JOURNAL OF RESEARCH AND APPLICATIONS IN AGRICULTURAL ENGINEERING, vol. 53, no. 4, 2009 - 2009, Poland, pages 125 - 130, XP055007439, Retrieved from the Internet <URL:http://www.pimr.poznan.pl/biul/2009_4_SKGS.pdf> [retrieved on 20110916] * |
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Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20140178528A1 (en) * | 2012-11-22 | 2014-06-26 | Instytut Biotechnologii Przemyslu Rolno-Spozywczego | Strain of lactobacillus buchneri a, composition, a multi-component preparation for starch-rich plant preservation, their use and a method for plant preservation |
US9370199B2 (en) * | 2012-11-22 | 2016-06-21 | Instytut Biotechnologii Przemyslu Rolno-Spozywczego | Strain of Lactobacillus buchneri A, composition, a multi-component preparation for starch-rich plant preservation, their use and a method for plant preservation |
WO2015014372A1 (en) * | 2013-07-30 | 2015-02-05 | OÜ Tervisliku Piima Biotehnoloogiate Arenduskeskus | Isolated microorganism strain lactobacillus plantarum tak 59 ncimb42150 and its use |
JP2016526900A (en) * | 2013-07-30 | 2016-09-08 | オウ テルヴィスリク ピーマ バイオテウノロジーテ アレンダスケスクス | Isolated microbial strain Lactobacillus plantarum TAK59 NCIMB42150 and uses thereof |
CN105861399A (en) * | 2016-06-17 | 2016-08-17 | 浙江大学 | Lactobacillus plantarum for preventing necrotic enteritis of chickens and application thereof |
CN105861399B (en) * | 2016-06-17 | 2019-11-05 | 浙江大学 | Lactobacillus plantarum and its application for preventing chicken necrotizing enterocolitis |
CN112143670A (en) * | 2020-09-08 | 2020-12-29 | 中国农业科学院农产品加工研究所 | Lactobacillus brevis 8-2B and application thereof |
CN112143670B (en) * | 2020-09-08 | 2022-07-19 | 中国农业科学院农产品加工研究所 | Lactobacillus brevis 8-2B and application thereof |
Also Published As
Publication number | Publication date |
---|---|
US20130084623A1 (en) | 2013-04-04 |
EP2582851B8 (en) | 2015-01-07 |
EP2582851A1 (en) | 2013-04-24 |
EP2582851B1 (en) | 2014-11-19 |
US8697423B2 (en) | 2014-04-15 |
PL214583B1 (en) | 2013-08-30 |
PL391534A1 (en) | 2011-12-19 |
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