EP3198040A1 - Probiotic fermented feed additives - Google Patents
Probiotic fermented feed additivesInfo
- Publication number
- EP3198040A1 EP3198040A1 EP15784768.2A EP15784768A EP3198040A1 EP 3198040 A1 EP3198040 A1 EP 3198040A1 EP 15784768 A EP15784768 A EP 15784768A EP 3198040 A1 EP3198040 A1 EP 3198040A1
- Authority
- EP
- European Patent Office
- Prior art keywords
- lab
- mscl
- feed
- baltbio02
- lactic acid
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Withdrawn
Links
Classifications
-
- 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
-
- 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
- A23K10/12—Animal feeding-stuffs obtained by microbiological or biochemical processes by fermentation of natural products, e.g. of vegetable material, animal waste material or biomass
-
- 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
- A23K10/35—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms from potatoes
-
- 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
-
- 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
Definitions
- the present invention relates to biotechnologies and is intended for the field of feed production and may be applied for producing preparations of high antimicrobial activity, i.e. fermented feed additives, feed supplements, feed premixtures, feed materials and compound feeds.
- Lactic acid bacteria are normally considered to be beneficial (nonpathogenic) microorganisms which live in various environments such as that of soil, plants, insects, animals and human microflora. These microorganisms are antagonistic to decay microflora and excrete a lactic acid as well as the substances which are identical to antibiotics, i.e. bacteriocins or substances similar to bacteriocins. Lactic acid bacteria promote the synthesis of B-group vitamins (biotin, thiamine, riboflavin, nicotinic acid, vitamin B12) and the vitamin K.
- B-group vitamins biotin, thiamine, riboflavin, nicotinic acid, vitamin B12
- lactic acid bacteria reaches the peak when the pH level of the media is 4-5.
- Their spectrum of activity includes many pathogenic bacteria: Escherichia Coli, Salmonella, Shigella, Pasteurella, Bacillus and Staphylococcus.
- Probiotics of lactic acid bacteria normalize the livestock gastrointestinal microflora, stimulate digestive processes, help their bodies to assimilate the nutrients contained in a feed, have influence on the livestock resistance to infections and their yield.
- Pediococcus spp. One of the most perspective genera of LAB is Pediococcus spp. and, in particular, two closely related group bacteria of this genus Pediococcus pentosaceus and Pediococcus acidilactici. Their brief morphological description is provided in the manual (Bergey's Manual of Systematic Bacteriology, vol.3, Furmicutes, Springer, 2009 p.p. 518, 519). The use of these bacteria in the production of fermented feed is known.
- the production method of dry (solid) fermented feeds is known (see the patent WO2013029682). It suggests to use one or several LAB from the group Enterococcus faecium, Lactobacillus rhamnosus, Lactobacillus plantarum, Pediococcus acidililactili and Pediococcus pentosaceus.
- the method and equipment of drying of fermented feeds is widely described in this invention.
- the disadvantage of this invention is high temperature necessary for drying fermented feeds: the temperature of the air supplied to the drying chamber exceeds +120°C, and the temperature of the dried fermented feeds exceeds +40°C; this significantly reduces the viability of probiotic bacteria.
- the homofermented liquid feeds and the methods of their production are also known (see the patent EP2056681). It suggests to use the same LAB in fermented feed production as the abovementioned invention, however, it additionally indicates specific LAB strains, i.e. Enterococcus faecium MCIMB 30122, Lactobacillus rhamnosus NCIMB 30121, Lactobacillus plantarum LSI (NCIMB 30083), Pediococcus acidililactili NCIMB 30086 and Pediococcus pentosaceus HTS (LMG P- 22549).
- the described method of production and formulas are not suitable for dry fermented feed production.
- the closest analogue for the proposed invention is described in the patent US20140037786. It is the method of dry fermented feed production in which such LAB strains as Enterococcus faecium MCIMB 30122, Lactobacillus rhamnosus NCIMB 30121, Lactobacillus plantarum LSI (NCIMB 30083), Pediococcus acidililactili NCIMB 30086 and Pediococcus pentosaceus HTS (LMG P-22549) are used.
- LAB strains as Enterococcus faecium MCIMB 30122, Lactobacillus rhamnosus NCIMB 30121, Lactobacillus plantarum LSI (NCIMB 30083), Pediococcus acidililactili NCIMB 30086 and Pediococcus pentosaceus HTS (LMG P-22549) are used.
- the objective of the invention is production of the fermented feed additives of high biological activity by using the newly selected LAB strains isolated from the spontaneously fermented cereal material. This objective is achieved by using, together or separately, the LAB Pediococcus acidilactici BaltBioOl and/or Pediococcus pentosaceus BaltBio02 in production of fermented feed additives.
- the BaltBio lactic acid bacteria strains are isolated from the starter cultures of the spontaneously fermented cereal crops grown in Lithuania. The technical works of isolation of the strains have been performed by UAB Bioseka under contracts. The strains have been identified by UAB Biosta.
- the starter cultures were produced under the laboratory conditions by changing the moisture content of the medium and the temperature of fermentation.
- the starter cultures were prepared from rye flour type 1370, oat flour and water in the ratio of 1 : 1 or 1:2, the temperature was cyclically varied within the range from 25 to 35 °C.
- the modified MRS medium (mMRS) was prepared by dissolving 10.0 g of tryptone peptone and meat extract, 5.0 g of yeast extract and sodium acetate, 7.0 g of glucose, fructose and maltose, 2.0 g of sodium glyconate, K2HPO4 and triammonium citrate, 1.0 ml of Tween 80, 0.2 g of MgS0 4 -7 H 2 0, 0.05 g of MnS0 4 -4 H 2 0, 0.5 g of cystein chloride, 15.0 g of agar in 1 1 of distilled water. The obtained solution (pH 6.3) was sterilized for 15 min at the temperature of 121 °C.
- the MRS agar medium (MRS) was prepared by dissolving 66.2 g of MRS agar in 1 1 of distilled water. The obtained solution was boiled until the agar was completely dissolved and sterilized for 15 min at the temperature of 118 °C.
- the LAB were characterized by identifying their morphology by performing the tests of carbohydrate fermentation and growth at different temperatures.
- Cocci were divided into two groups on the basis of sucrose and maltose fermentation: the first group consisted of 26, and the second group consisted of 10 bacterial strains. All spherical- shaped bacteria hydrolyzed trehalose and glucose. It is known that P. pentosaceus and P. acidilactici differ in fermentation of maltose and trehalose and their potential for growth is also different at the temperature of 50°C, P. pentosaceus ferment both carbohydrates and are unable to grow at the said temperature, while P. acidilactici ferment only trehalose and are able to grow at the temperature of 50 °C.
- bacteriocins of the isolated strains P. acidilactici and P. pentosaceus against pathogenic microorganisms Bacillus thuringiensis, Escherichia coli, Enterococcus faecalis, Staphylococcus aureus, Bacillus macerans, Salmonella enteritidis, Micrococcus sp, Yersinia enterocolitica, Listeria sp., Pseudomonas aeroginosa), Pediococcus acidilactici BaltBioOl and Pediococcus pentosaceus BaltBio02 were selected as the most promising microorganisms for ensuring good health of animals by using them for feed additives and other feed products.
- the antimicrobial activity of metabolites of the selected lactic acid bacteria P. acidilactici BaltBioOl, P. pentosaceus BaltBio02 against pathogenic bacteria (Bacillus thuringiensis, Escherichia coli, Enterococcus faecalis, Staphylococcus aureus, Bacillus macerans, Salmonella enteritidis, Micrococcus sp, Yersinia enterocolitica, Listeria sp., Pseudomonas aeroginosa) has been found.
- LAB lactic acid bacteria
- LAC are able to completely stop the growth of pathogenic bacteria (bactericidal effect) or inhibit or partly inhibit their growth (bacteriostatic effect).
- the antimicrobial effect of the LAC strains against Gram-/+ bacteria depended on the LAB strain and indicator microorganism.
- P. acidilactici BaltBioOl demonstrated the bactericidal effect against all examined indicator bacteria (measured diameter of the zones of inhibition in the agar medium varied from 10 ⁇ 0 to 24.5 ⁇ 0.7 mm), while P. pentosaceus BaltBio02 demonstrated the bacteriostatic effect.
- the LAB metabolites demonstrated fungistatic and fungicidal effect on the growth of Fusarium poae, while the LAB had only fungistatic effect on the growth of Penicillium chrysogenum and Aspergillus niger.
- LAB Prior to experiment, LAB were kept at the temperature of -70°C (Sanyo, Japan). The thawed LAB were amplified in MRS agar (Sigma Aldrich, Germany) or any other alternative medium by keeping P. pentosaceus BaltBio02 at the temperature of 35 °C and P. acidilactici BaltBioOl - at the temperature of 32 °C for 48 hours, until the level of LAB reached 10 8 -10 9 CFU/ml.
- the pure LAB are amplified in the alternative medium which is made of potato juice by autoclaving them, cooling to a temperature of 30°C and introducing pure LAB cultures (1 ml pure bacteria cultures amplified in MRS bouillon into 0.5 litre of juice).
- the alternative medium with LAB is kept at the temperature of 30 +/- 2 °C for 24; 48 and 72 hours, until the level of LAB reaches 9.60xl0 10 CFU/ml.
- LAB prepared for fermentation may comprise from 0.5% to 10% of the substratum prepared before fermentation.
- P. acidilactici BaltBioOl or P. pentosaceus BaltBio02 or their mixture may be used.
- the recommended duration of fermentation is from 8 hours to 72 hours and the temperature shall be from 20°C to 40°C.
- Biologically active fermented feed additive shall contain LAB from 10 6 to 10 9 CUF/g and pH shall be from 3.0 to 6.0.
- Example 1 Assessment of the Effect of the Product Fermented by the Mixture of Lactic Acid Bacteria Pediococcus Acidilactici BaltbioOl and Pediococcus Pentosaceus Baltbio02 on the Health Status of Calves.
- Intense calf rearing which is now widely applied, causes gut microflora imbalance, since the appropriate microflora, which is formed from the environment and is obtained with maternal milk in the gastrointestinal tract of the livestock reared under the natural conditions, fails to develop in the gastrointestinal tract of the intensively reared livestock. For various reasons, changes in the composition of microorganisms of small intestine result in growth of coliform baccili which replace the lactic acid bacteria.
- the LAB materials are able to normalize the gut microflora, improve the defensive features of the mucosa, promote nutrient assimilation and natural resistance as well as reduce the stress-induced consequences.
- milk composition fat content %, protein content % , lactose %, somatic cell count, (SCC) thousand/ml), urea, (mg/per cent).
- SCC somatic cell count
- urea urea
- Example 3 Milk Yield and Chemical Composition when Feeding Cows with the Pediococcus pentosaceus Additive. Methodology of the Experiment of Feeding Lithuanian Black-and- White Cows with the LAB Pediococcus pentosaceus BaltBio02 Amplified and Prepared for Fermentation.
- the experimental cows were fed with the same ration. However, in addition to the feed, these cows were given 100 g of the experimental additive consisting of Pediococcus pentosaceus
- the milk yield was measured three times, i.e. at the beginning of the experiment, in the middle of the experiment (after a half of the experimental period) and at the end of the experiment by carrying out the control milkings.
- the quantity of milk with natural fat content was recalculated into the quantity of 4 per cent fat (corrected) milk on the basis of the respective ratios.
- Milk samples were taken from each cow individually in accordance with guidance on milk sampling (LST EN ISO 707: 1999+P:2003 Milk and milk products. Guidance on sampling) at the beginning, in the middle and at the end of the experiment. Milk fat content, milk protein content, lactose and urea were measured by using the device "LactoScope FTIR" (FT1.0.
- the LAB strains P. acidilactici BaltBioOl and P. pentosaceus BaltBio02 which have been isolated from the Lithuanian cereal raw material and tested, are microbiologically safer, have high biological activity and improve the health status of animals, increase their yield and may be successfully used in fermented feed production.
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- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Polymers & Plastics (AREA)
- Biotechnology (AREA)
- Health & Medical Sciences (AREA)
- Zoology (AREA)
- Microbiology (AREA)
- Food Science & Technology (AREA)
- Animal Husbandry (AREA)
- Physiology (AREA)
- Molecular Biology (AREA)
- Wood Science & Technology (AREA)
- Biomedical Technology (AREA)
- Biochemistry (AREA)
- Genetics & Genomics (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Organic Chemistry (AREA)
- General Engineering & Computer Science (AREA)
- Botany (AREA)
- Mycology (AREA)
- General Health & Medical Sciences (AREA)
- Virology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Medicinal Chemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Sustainable Development (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
- Fodder In General (AREA)
Abstract
Description
Claims
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
LT2014109A LT6270B (en) | 2014-09-24 | 2014-09-24 | Probiotic fermented feed additives |
PCT/IB2015/057168 WO2016046706A1 (en) | 2014-09-24 | 2015-09-17 | Probiotic fermented feed additives |
Publications (1)
Publication Number | Publication Date |
---|---|
EP3198040A1 true EP3198040A1 (en) | 2017-08-02 |
Family
ID=54347594
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
EP15784768.2A Withdrawn EP3198040A1 (en) | 2014-09-24 | 2015-09-17 | Probiotic fermented feed additives |
Country Status (4)
Country | Link |
---|---|
EP (1) | EP3198040A1 (en) |
EA (1) | EA201790685A1 (en) |
LT (1) | LT6270B (en) |
WO (1) | WO2016046706A1 (en) |
Families Citing this family (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
LT6271B (en) | 2014-09-24 | 2016-05-10 | Uab "Baltijos Biotechnologijos" | Fermented feed of plant origin |
LT6480B (en) | 2017-04-10 | 2017-12-11 | Aleksandro Stulginskio Universitetas | Dehydrated meadow grass meal |
WO2018224420A1 (en) | 2017-06-06 | 2018-12-13 | Chr. Hansen A/S | A pediococcus acidilactici strain capable of increasing milk yield |
CN109439565A (en) * | 2018-09-14 | 2019-03-08 | 湖南师范大学 | One plant of Pediococcus pentosaceus bacterial strain, its probiotics and preparation method thereof |
CN111406856B (en) * | 2020-03-11 | 2023-04-07 | 天津科技大学 | Functional fermented beverage for dispelling effects of alcohol and protecting liver and preparation method thereof |
CN112159460B (en) * | 2020-09-29 | 2021-12-17 | 厦门大学 | Scylla paramamosain antibacterial peptide Spamprin4-23And uses thereof |
CN118185811B (en) * | 2024-03-13 | 2024-08-02 | 广东海洋大学 | Fermented feed rich in antibacterial peptide, preparation method and application |
Family Cites Families (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
SU378405A1 (en) * | 1971-01-07 | 1973-04-18 | Всесоюзный научно исследовательский институт биосинтеза белковых веществ | METHOD OF CULTIVATION OF MICROORGANISMSI2 |
US20050130288A1 (en) * | 2003-12-11 | 2005-06-16 | Eromlife Co., Ltd. | Novel microorganism Pediococcus pentosaceus EROM101, having immune enhancement, anticancer and antimicrobial activities |
JP5438510B2 (en) | 2006-07-14 | 2014-03-12 | ローン レガース | Homo fermentation product |
EP2688419A1 (en) | 2011-03-22 | 2014-01-29 | Fermentationexperts AS | Improved fermentation of protein-rich feed |
WO2013029682A1 (en) | 2011-09-01 | 2013-03-07 | Schaeffler Technologies AG & Co. KG | Drive device |
CN103173373B (en) * | 2012-07-12 | 2015-05-20 | 北京伟嘉人生物技术有限公司 | Pediococcus acidilactici with high bacteriostatic activity and application thereof |
CN103156091B (en) * | 2013-03-20 | 2014-09-10 | 哈尔滨市乳家科技饲料有限公司 | Method for manufacturing biologically-fermented straw nutrition forage grass at low temperature for beef |
-
2014
- 2014-09-24 LT LT2014109A patent/LT6270B/en not_active IP Right Cessation
-
2015
- 2015-09-17 EA EA201790685A patent/EA201790685A1/en unknown
- 2015-09-17 WO PCT/IB2015/057168 patent/WO2016046706A1/en active Application Filing
- 2015-09-17 EP EP15784768.2A patent/EP3198040A1/en not_active Withdrawn
Non-Patent Citations (2)
Title |
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None * |
See also references of WO2016046706A1 * |
Also Published As
Publication number | Publication date |
---|---|
LT6270B (en) | 2016-05-10 |
LT2014109A (en) | 2016-04-11 |
EA201790685A1 (en) | 2017-07-31 |
WO2016046706A1 (en) | 2016-03-31 |
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