IE59968B1 - Improvements in or relating to acidifying green fodder - Google Patents

Improvements in or relating to acidifying green fodder

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Publication number
IE59968B1
IE59968B1 IE124787A IE124787A IE59968B1 IE 59968 B1 IE59968 B1 IE 59968B1 IE 124787 A IE124787 A IE 124787A IE 124787 A IE124787 A IE 124787A IE 59968 B1 IE59968 B1 IE 59968B1
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IE
Ireland
Prior art keywords
lactobacillus plantarum
dsm
green fodder
silage
plantarum dsm
Prior art date
Application number
IE124787A
Other versions
IE871247L (en
Original Assignee
Sanofi Ceva Gmbh
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by Sanofi Ceva Gmbh filed Critical Sanofi Ceva Gmbh
Publication of IE871247L publication Critical patent/IE871247L/en
Publication of IE59968B1 publication Critical patent/IE59968B1/en

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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
    • 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
    • C12R2001/225—Lactobacillus
    • C12R2001/25—Lactobacillus plantarum

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  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Health & Medical Sciences (AREA)
  • Zoology (AREA)
  • Microbiology (AREA)
  • Wood Science & Technology (AREA)
  • Biotechnology (AREA)
  • Polymers & Plastics (AREA)
  • Organic Chemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Genetics & Genomics (AREA)
  • Virology (AREA)
  • General Chemical & Material Sciences (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Biomedical Technology (AREA)
  • Animal Husbandry (AREA)
  • Medicinal Chemistry (AREA)
  • Food Science & Technology (AREA)
  • Fodder In General (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
  • Medicines Containing Material From Animals Or Micro-Organisms (AREA)
  • Storage Of Fruits Or Vegetables (AREA)

Abstract

Process for ensiling green fodder using bacterial preparations of lactic acid-producing bacteria, in which Lactobacillus plantarum DSM 3676 and/or Lactobacillus plantarum DSM 3677 is used.

Description

The invention relates to a process for acidifying green fodder by using bacterial preparations of lactic acid-producing bacteria.
Silage making from green fodder has enormous economic 5 importance, and various processes and additives have been proposed for accelerating the fermentation of green fodder batches or for improving silage making. These include the addition of lactic acid-producing bacteria, because the fermentation of lactic acid is an essential process in silage making from green fodder.
At the end of fermentation, a good silage should contain about 2 to 3% lactic acid by weight. The bacteria added should be able to ferment all or as much as possible of the sugar contained in the green fodder. But this aim has not been adequately achieved by the known bacteria which have previously been added to silage (cf OE~A 252 040).
« The problem which the present invention seeks to solve, therefore, is to provide bacterial preparations or bacteria which yield improved silage, or which in particular effect a stronger acidification in a quicker time, while substantially avoiding a butyric acid fermentation. - 3 The invention accordingly provides a process for the acidification of green fodder using bacterial preparations of lactic acid-producing bacteria, which is characterized by the use of Lactobacillus plantarum DSM 3675 and/or t ι Lactobacillus plantarum DSM 3677.
It has surprisingly been found that the two microorganisms DSM 3676 and DSM 3677, when used together, have a synergistic action. It is therefore preferred to use a mixture of 10 to 90% Lactobacillus plantarum DSM 3675 and 90 to 10% Lactobaci11 us plantarum DSM 3677, referred to the A mixture of 30 to 70% DSM 3676 especially 40 to 60% DSM 3676 and particularly preferred. number of cells present and 70 to 30% DSM 3677, 60% to 40% DSM 3577, is The invention relates also to the use of Lactobacillus plantarum DSM 3676 and/or Lactobacillus plantarum DSM 3677 for the ensiling of green fodder.
The two microorganisms mentioned above, used in accordance with the invention, have not been described heretofore.
One of these microorganisms was originally labelled by the applicant with the designation B5 and deposited on 12 March 1986 in the German collection of microorganisms, Grisebachstr 8, D-3400 Goettingen, and received the deposit number DSM 3676. (Deposit according to the Budapest Treaty on the internetional recognition of deposits of microorganisms for the purposes of patent procedures).
The second microorganism was originally labelled by the applicant with the designation W2 and likewise deposited on , 12 March 1986 at the same depository. It was accorded the deposit number DSM 3677. » Both microorgan isms were identified as Lactobaci11 us plantarum, subspecies arabinosus. They exhibit the following properties: . 4 no gas formation DL - lactic acid no ammonia from arginine optimum temperature c 35C no growth at 45’C growth at 15"C cell wall component: diami nopimo1inic acid sugar fermentation g1ucose mannitol g1 veer i n arab i nose sucrose pos i t ive positive negative pos i t ive pos i t ive 15 1actose positive mel1ibiose positive dextrin positive ( in the case of W2) (weaker in the case of B5) starch negat i ve 20 esculin (? illegible) pos itive maltose positive melicitose positive xylose negative ribose positive 25 t r i h a 1 o s e pos it ive raffinose positive sorbitol positive salicinose pos i t i ve rhamnose positive (in the case of W2) 30 (negative in the case of B5) Differences: W2 forms mucus B5 forms no mucus - 5 Acetate tolerance: 1.5% for W2 0.5% for B5 Morphology/: W2 = single rods B5 = short rods in chains Acidification in MRS pH 3.81 for B5 pH 3.79 for W2 Metabolism determined for B5: 2 mol lactate are formed from one mol glucose, 1 mol lactate plus 0.65 mol acetate and traces of ethanol are formed from 1 mol dripose.
The microorganisms can be cultivated in conventional manner. Suitable culture conditions in MRS medium: pH before sterilization: 6.9 steri1ization 20 minutes at 121’C pH after steri1ization : 6.6 behaviour towards oxygen: microaerophi1ic (1) On preservative agar with refrigerator storage.
Incubation temperature: 25 to 32°C Duration of incubation: 24 hours Sub-cultivation interval: about 3 months.
Suitable storage conditions: a) master culture in preservative agar (conventional for Lactobacilli). After initial growth, storage in / refrigerator. b) Freeze-dried culture has almost unlimited storage life * at -20’C.
Conditions for viability testing: Poured plate or master culture in MRS-media.
The new microorganisms are characterized by special capabilities in the intensity and speed of fermentation of sugars, such as are present in green fodder, at normal silage temperatures.
The microorganisms were isolated from green fodder silage in the early phase of fermentation.
The following tables describe the results achieved during acidification of grass or lucerne using the microorgan isms according to the invention, and silage media of the present state of the art. The lower the pH value, the stronger is the acidification achieved, and the better are the results for the s i1 age.
The following abbreviations are used in the tables: TS: dry substance Kofa-Lac (registered trade mark): commercial state-of-theart preparation Kofa-Plus: commercial state-of-the-art preparation based on chemicals 5 5 The numbers 10 or 10 represent the numbers of cells per 1 ml or 1 g of the fresh batch (inoculation strength). Unster: unsterile Biomax (registered trade mark): commercial state-of-theart preparation The figures given in the tables are the respective pH values.
Table 1. Acidification speed (drop in pH) in sterile and unsterile, artificial (MRS-A to C according to inoculation strength) and natural media (2 batches) at 20" and 30°C. 6 Inoculation strength per 10 ml: 5 x 10 - 5 x 10 cells.
Test No Agent from Test No Nutrient medium Grass + lucerne unster. maize A unster.
MRS/A sterile MRS/B sterile Grass + lucerne unster Maize + lucerne sterile| Maize B unster.
Maize B sterile MRS/C sterile °C/24h I 20"C/24h Ί- 1 pH I 6.28 "T" 1 5.78 T 1 6.49 1 1 6.48 1 T 1 6.34 1 1 1 6.41 1 ! 1 5.81 (sterile, 0 1 1 1 1 1 1 1 1 1 1 1 1 1 Ί |Kofa-Lac | 5.22 1 1 5.43 1 1 5.65 1 1 6.00 1 6.00 1 1 5.80 1 1 5.48 1 |Biomax j I 5.87 1 1 I 5.52 1 1 I 6.26 1 1 I 1 1 I 1 1 I 1 1 I |W 2 j I 4.98 1 1 4.37 1 1 5.31 1 1 5.82 1 1 5.39 1 1 5.70 1 1 I 4.67 |B 5 | 4.51 1 1 4.23 1 1 4.75 1 1 4.97 1 1 4.95 1 1 5.05 1 1 4.54 I 6.02 6.49 |Kofa-Lac I Biomax I · |W 2 I IB 5 31 1 3.51 1 4.02 1 4.00 | | I 4.28 38 1 3.59 1 4.11 1 1 1 I 1 1 I 16 1 3.47 1 3.97 1 3.99 | 1 4.22 33 1 3.88 1 4.19 1 4.18 | I 4.33 unster non-sterile .41 .22 .21 6.33 6.00 4.95 .70 .27 4.80 .80 .40 4.94 ,43 .88 4.98 4.50 3.98 3.99 4.18 4.02 3.97 4.18 3.95 4.03 3.87 4.13 4.09 4.07 4.23 Table 2 Grass TS ca- 18% Ts, Week glass 22°C and 30°C. pH development in days Variables 3O’C 1 1 1 0 | 1 1 1 1/2 1 1 4 1 0 1 1 1 1 5.21 1 I 4.72 (Kofa-Lac 1 1 4.64 I 4.46106 (85 I 5.68 t 4.54 J VC (W2 1 1 | 1 4.53 1 4.41 1 (Kofa-L ac 1 1 1 1 4.73 1 1 4.62 IO5 (B5 f 1 4.69 I 4.50 (W2 1 1 | 1 4.56 I 4.51 I Kof a~P 1 us 1 1 1 1 1 1 4.84 I 1 4.45 1 ★ w » .41 | 4 .48 | 4 .48 .76 .35 ic 22'C not yet opened fermentation gas losses 1.65 - 2.25 g/vessel. no confirmed differences in inoculation variants.
Table 3 Acidification in pressed liquor (pH measured directly) Grass TS c 18% Pressed liquor 1 1 1 30eC | 22°C 1 21b | 45h | 4d | 21h I 45 h I 4d 1 1 0 1 1 ! 4.71 | 4.04 1 I 4.05 1 I 5.67 1 | 4.44 1 I 4.13 | (Kofa-Lac I 4.07 | 3.95 I 3.93 | 5.65 I 4.20 I 4.12 I 106 (B5 I 3.95 | 3.95 | 3.93 | 4.84 | 4.15 i 4.10 10 1 (W2 I 4.02 | 3.97 I 3.97 I 4.95 | 4.12 | 4.09 ί 1 I (Kofa-Lac 1 1 1 4.28 | 4.00 1 I 3.98 1 1 1 1 1 1 1 105 (Bg I 4.02 | 3.97 I 3.93 1 1 1 1 (W2 1 I 4.03 | 1 1 4.00 I 3.98 1 1 1 1 1 1 1 - ίο Table 4 Acidification in pressed liquor Fresh lucerne Pressed liquor ’C 22°C 1 Id | 1 1 2d 0 1 1 1 5.18 | 4.76 (Kofa-lac | 4.46 | 4.44 106 (B5 | 4.35 | 4.30 (W2 | 4.41 | 4.36 (RP I 4.57 1 1 1 4.41 (Kofa-L ac 1 1 | 4.70 1 4.61 105 (B5 | 4.52 | 4.48 (W2 | 4.50 | 4.48 4d | 1 Id | 2d 1 | 4d 1 1 4.76 | 5.95 1 | 5.15 1 I 5.03 4.43 | 5.91 | 4.80 I 4.79 4.30 t 5.61 I 4.55 t 4,53 4.35 | 5.69 | 4.60 I 4.60 4.40 | 1 I 1 1 4.66 | 1 1 1 1 4.47 | 1 1 4.47 | 1 1 The cultivation of the bacteria can be carried out in conventional manner (cf OE-A-262 040). If necessary a substance having an alkaline reaction can be added to the nutrient medium during cultivation, in order to prevent the pH value from dropping too low. Examples of such alkaline substances are potassium hydroxide solution, sodium hydroxide solution, soda solution. The separation of the cells can be carried out in conventional manner. For example the cells can be separated in a centrifuge. A protective substance, for example a protective colloid, eg powdered milk can then be added to them. Other protective substances can also be used, for example lactose, ascorbic acid.
The cells can then be freeze-dried. A stable preparation is obtained, which contains the active bacteria and can be stored for a long time without losing its activity.
The preparation can be added to the silage mix in known manner by hand, or by the use of metering devices, powder sprays and the like.
US-A-4 528 199 and AT-A-262 040 describe use of Lactobacillus plantarum preparations for ensiling green xodder. As compared with the preparations described therein, the bacteria used according to the invention have unexpectedly improved properties with regard to the speed of ensiling.

Claims (8)

1. C1 aims r 1. A process for acidifying green fodder using . preparations of lactic acid-producing bacteria, characterized in that Lactobacillus plantarum DSM 3676 5 and/or Lactobacillus plantarum DSM 3677 is/are used.
2. A process according to claim 1, characterized in that a mixture of 10 to 90% Lactobacillus plantarum DSM 3676 and 90 to 10% Lactobacillus plantarum OSH 3677, based on the number of cells, is used. 10
3. A process according to claim 1, characterized in that a mixture of 30 to 70% Lactobacillus plantarum DSM 3676 and 70 to 30% Lactobacillus plantarum DSM 3677, based on the number of cells, is used.
4. Use of Lactobacillus plantarum DSM 3676 and/or 15 ' Lactobacillus plantarum DSM 3677 for making silage from green fodder.
5. A biologically pure culture of Lactobacillus plantarum ' Q DSM 3676.
6. » a biologically pure culture of Lactobacillus plantarum 20 DSM 3677.
7. A process for acidifying green fodder, substantially as herein described with reference to any of tables 1 to 4.
8. Silage whenever produced from green fodder acidified by a process claimed in a preceding claim.
IE124787A 1986-05-14 1987-05-13 Improvements in or relating to acidifying green fodder IE59968B1 (en)

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
DE19863616181 DE3616181A1 (en) 1986-05-14 1986-05-14 PROCESS FOR ACIDIFYING GREEN FORAGE

Publications (2)

Publication Number Publication Date
IE871247L IE871247L (en) 1987-11-14
IE59968B1 true IE59968B1 (en) 1994-05-04

Family

ID=6300784

Family Applications (1)

Application Number Title Priority Date Filing Date
IE124787A IE59968B1 (en) 1986-05-14 1987-05-13 Improvements in or relating to acidifying green fodder

Country Status (5)

Country Link
EP (1) EP0250786B1 (en)
AT (1) ATE80978T1 (en)
DE (2) DE3616181A1 (en)
IE (1) IE59968B1 (en)
NO (1) NO169573C (en)

Families Citing this family (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
FI82354C (en) * 1988-11-14 1991-03-11 Valio Meijerien Preservation of fresh food
DE3916563A1 (en) * 1989-05-20 1990-11-22 Atochem Werke Gmbh COMBINATION DEVICE AND METHOD FOR ACIDIFYING GREEN FORAGE AND PREVENTING AEROBIC DEGRADING PROCESSES IN GAERFUTTER
DE4034749C2 (en) * 1990-11-01 2002-11-07 Addcon Agrar Gmbh Combination preparation and method for acidifying green fodder and preventing aerobic degradation processes in fermented fodder
DE4112866A1 (en) * 1991-04-19 1992-10-22 Sanofi Ceva Gmbh Inhibiting Listeria in fermented fodder, esp. silage - by adding alkali salt of sulphurous acid and nitrite giving synergistic effect
ES2559842T3 (en) * 2008-07-21 2016-02-16 Erber Aktiengesellschaft Procedure for the treatment of forage silage for ruminants, as well as additive for forage silage

Family Cites Families (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
AT262040B (en) * 1964-07-13 1968-05-27 Theodor Dr Beck Process for the production of a lactic acid bacterial preparation
IT1109471B (en) * 1976-08-17 1985-12-16 Deral Sa PROCEDURE AND PRODUCT FOR THE PRESERVATION AND ENHANCEMENT OF GREEN VEGETABLES AND OF THE WET PRODUCTS UNDER AGRO-FOOD INDUSTRIES
US4528199A (en) * 1983-01-26 1985-07-09 University Of Georgia Research Foundation, Inc. Silage production from fermentable forages
FR2542013B1 (en) * 1983-03-01 1986-01-03 Abc Bio Ind PROCESS FOR THE PREPARATION AND STORAGE OF A PROTEIN HYDROLYSAT USEFUL IN PARTICULAR IN THE AGRI-FOOD FIELD

Also Published As

Publication number Publication date
DE3781943D1 (en) 1992-11-05
DE3616181A1 (en) 1987-11-26
NO169573C (en) 1992-07-15
NO871988D0 (en) 1987-05-13
ATE80978T1 (en) 1992-10-15
IE871247L (en) 1987-11-14
NO169573B (en) 1992-04-06
EP0250786B1 (en) 1992-09-30
EP0250786A2 (en) 1988-01-07
NO871988L (en) 1987-11-16
EP0250786A3 (en) 1989-07-05

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