CN111876365B - Lactobacillus rhamnosus and application thereof - Google Patents
Lactobacillus rhamnosus and application thereof Download PDFInfo
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- CN111876365B CN111876365B CN202010951401.0A CN202010951401A CN111876365B CN 111876365 B CN111876365 B CN 111876365B CN 202010951401 A CN202010951401 A CN 202010951401A CN 111876365 B CN111876365 B CN 111876365B
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- 241000218588 Lactobacillus rhamnosus Species 0.000 title claims abstract description 29
- 238000000855 fermentation Methods 0.000 claims abstract description 24
- 230000004151 fermentation Effects 0.000 claims abstract description 24
- 241001465754 Metazoa Species 0.000 claims abstract description 10
- 238000004321 preservation Methods 0.000 claims abstract description 9
- 230000012010 growth Effects 0.000 claims abstract description 7
- 230000001580 bacterial effect Effects 0.000 claims abstract description 4
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 6
- 239000001963 growth medium Substances 0.000 claims description 6
- 239000007788 liquid Substances 0.000 claims description 6
- 229930006000 Sucrose Natural products 0.000 claims description 3
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 claims description 3
- 230000001737 promoting effect Effects 0.000 claims description 3
- 239000011780 sodium chloride Substances 0.000 claims description 3
- 239000005720 sucrose Substances 0.000 claims description 3
- 230000001502 supplementing effect Effects 0.000 claims description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 3
- 230000000813 microbial effect Effects 0.000 claims description 2
- 238000009629 microbiological culture Methods 0.000 claims description 2
- 230000001954 sterilising effect Effects 0.000 claims description 2
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims 1
- 238000012136 culture method Methods 0.000 claims 1
- 239000000843 powder Substances 0.000 claims 1
- 210000000936 intestine Anatomy 0.000 abstract description 9
- 210000002784 stomach Anatomy 0.000 abstract description 9
- 238000000034 method Methods 0.000 abstract description 6
- 238000009825 accumulation Methods 0.000 abstract description 3
- 244000052616 bacterial pathogen Species 0.000 abstract description 3
- 230000006870 function Effects 0.000 abstract description 3
- 239000002207 metabolite Substances 0.000 abstract description 3
- 230000000644 propagated effect Effects 0.000 abstract description 3
- 108020004465 16S ribosomal RNA Proteins 0.000 abstract description 2
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 description 6
- 241000894006 Bacteria Species 0.000 description 5
- 239000003814 drug Substances 0.000 description 5
- 241000238553 Litopenaeus vannamei Species 0.000 description 4
- 229940079593 drug Drugs 0.000 description 4
- 241000186660 Lactobacillus Species 0.000 description 3
- 239000003242 anti bacterial agent Substances 0.000 description 3
- 229940088710 antibiotic agent Drugs 0.000 description 3
- 201000010099 disease Diseases 0.000 description 3
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 3
- 239000005556 hormone Substances 0.000 description 3
- 229940088597 hormone Drugs 0.000 description 3
- 235000014655 lactic acid Nutrition 0.000 description 3
- 239000004310 lactic acid Substances 0.000 description 3
- 239000001888 Peptone Substances 0.000 description 2
- 108010080698 Peptones Proteins 0.000 description 2
- 238000009395 breeding Methods 0.000 description 2
- 230000001488 breeding effect Effects 0.000 description 2
- 230000007547 defect Effects 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 230000036541 health Effects 0.000 description 2
- 230000006872 improvement Effects 0.000 description 2
- 229940039696 lactobacillus Drugs 0.000 description 2
- 235000019319 peptone Nutrition 0.000 description 2
- 208000031295 Animal disease Diseases 0.000 description 1
- 241000238557 Decapoda Species 0.000 description 1
- 241000192125 Firmicutes Species 0.000 description 1
- 240000001046 Lactobacillus acidophilus Species 0.000 description 1
- 235000013956 Lactobacillus acidophilus Nutrition 0.000 description 1
- 244000199866 Lactobacillus casei Species 0.000 description 1
- 235000013958 Lactobacillus casei Nutrition 0.000 description 1
- 241000186605 Lactobacillus paracasei Species 0.000 description 1
- 240000006024 Lactobacillus plantarum Species 0.000 description 1
- 235000013965 Lactobacillus plantarum Nutrition 0.000 description 1
- 241000194020 Streptococcus thermophilus Species 0.000 description 1
- 230000008827 biological function Effects 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- 235000014633 carbohydrates Nutrition 0.000 description 1
- 238000000354 decomposition reaction Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000003912 environmental pollution Methods 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 239000013505 freshwater Substances 0.000 description 1
- 210000001035 gastrointestinal tract Anatomy 0.000 description 1
- 229940039695 lactobacillus acidophilus Drugs 0.000 description 1
- 229940017800 lactobacillus casei Drugs 0.000 description 1
- 229940072205 lactobacillus plantarum Drugs 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 230000021332 multicellular organism growth Effects 0.000 description 1
- 230000035790 physiological processes and functions Effects 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 239000010865 sewage Substances 0.000 description 1
- 241000894007 species Species 0.000 description 1
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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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- 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
- A23K50/00—Feeding-stuffs specially adapted for particular animals
- A23K50/80—Feeding-stuffs specially adapted for particular animals for aquatic animals, e.g. fish, crustaceans or molluscs
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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- C12N1/20—Bacteria; Culture media therefor
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/11—Lactobacillus
- A23V2400/175—Rhamnosus
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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
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Abstract
The invention relates to a Lactobacillus rhamnosus strain and application thereof, wherein the strain Lactobacillus rhamnosus (Lactobacillus rhamnosus) has a preservation number of CGMCC No.20472, and a 16s rDNA base sequence of the strain is shown as SEQ ID NO: 1 is shown. The invention also relates to a method for using the strain in cultivation, which comprises the following specific steps: (1) a fermentation medium formula; (2) fermentation processes (including fermentation parameters and methods); (3) a fermentation liquor cultivation application method; the invention has the advantages that the bacterial strain enters the intestines and stomach of the animal along with the feed, is rapidly propagated in the intestines and stomach and occupies the micro-ecological advantage, inhibits the propagation of pathogenic bacteria, avoids the accumulation of harmful metabolites, improves the functions of the intestines and stomach and promotes the growth of the animal.
Description
Technical Field
The invention relates to a strain and application thereof, in particular to lactobacillus rhamnosus and application thereof.
Background
At present, most farmers generally pay attention to improvement of feeding conditions and improvement of feeding level, but hopefully use animal health for medication, medication in case of diseases and medication in case of no diseases, so that the dosage of antibiotics and hormones far exceeds the requirement of animal disease treatment, drug-resistant bacteria are increased, diseases are difficult to control, feeding cost is increased, and product quality is reduced. And products using excessive antibiotics and hormones can also harm human health, and the excessive antibiotics and hormones flow into the environment along with excrement and sewage to cause environmental pollution.
Lactic acid bacteria are a general term for a group of bacteria that can produce large amounts of lactic acid using fermentable carbohydrates. The bacteria are widely distributed in nature and have abundant species diversity. Except for a very small number, most of them are flora which are indispensable in human body and have important physiological functions, and are widely present in intestinal tracts of human body. The lactobacillus has extremely high application value in important fields closely related to human life, such as industry, agriculture and animal husbandry, food, medicine and the like. However, the lactobacillus used in the breeding industry is mainly focused on lactobacillus acidophilus, lactobacillus casei, lactobacillus paracasei, lactobacillus plantarum, streptococcus thermophilus, etc. and more excellent lactobacillus species with special biological functions need to be developed, which is limited by research level and production difficulty.
Disclosure of Invention
The invention aims to solve the defects of the prior art and provides lactobacillus rhamnosus.
The second purpose of the invention is to solve the defects of the prior art and provide the application of lactobacillus rhamnosus, the lactobacillus rhamnosus strain (LR-003) is added into the feed, enters the intestines and stomach of the animal along with the feed, is rapidly propagated in the intestines and stomach and occupies microecological advantages, inhibits the propagation of pathogenic bacteria, avoids the accumulation of harmful metabolites, improves the functions of the intestines and stomach and promotes the growth of the animal.
In order to achieve the purpose, the invention adopts the following technical scheme:
a Lactobacillus rhamnosus (Lactobacillus rhamnosus) strain, strain code: LR-003, the preservation number is CGMCC No.20472, the preservation unit is China general microbiological culture Collection center (CGMCC), the preservation unit address is: the microbial research institute of the national academy of sciences No. 3, Xilu No. 1, Beijing, Chaoyang, and the preservation date is 07-30 days in 2020.
The strain is gram-positive bacteria, physiological, biochemical and 16rDNA sequence determination shows that the strain LR-003 is Lactobacillus rhamnosus (Lactobacillus rhamnosus), and the 16s rDNA base sequence of the strain is shown as SEQ ID NO: 1 is shown.
The invention also discloses a method for applying the lactobacillus rhamnosus strain to the breeding industry, which comprises the following steps:
1. preparing a culture medium: 1% (g/ml) of sucrose, 2% (g/ml) of peptone, 0.5% (g/ml) of sodium chloride, supplementing water, and sterilizing for later use;
2. liquid submerged fermentation culture: inoculating the single bacterial colony of the strain into the culture medium, performing shake culture at 100r/min, wherein the fermentation temperature is 37 ℃, the fermentation time is 36 hours, and the OD value of the fermentation liquid is 5;
3. application for promoting animal growth: the lactobacillus rhamnosus fermentation liquor needs to be mixed with feed for feeding, wherein the usage amount of the fermentation liquor is 1 per mill (mL/g) of the feed.
The invention has the advantages that: the strain LR-003 is lactic acid bacteria, has good safety and strong decomposition capability, enters the intestines and stomach of animals along with feed, is rapidly propagated in the intestines and stomach and occupies microecological advantages, inhibits the propagation of pathogenic bacteria, avoids the accumulation of harmful metabolites, improves the functions of the intestines and stomach, and promotes the growth of the animals. The method is simple and easy to implement, stable in effect and environment-friendly.
Drawings
FIG. 1 is a diagram: a growth promoting effect diagram of the strain of lactobacillus rhamnosus CGMCC No.20472 for Penaeus vannamei Boone.
Detailed Description
Example 1:
an application of Lactobacillus rhamnosus (Lactobacillus rhamnosus) strain:
a. preparing a culture medium: 1% (g/ml) of sucrose, 2% (g/ml) of peptone, 0.5% (g/ml) of sodium chloride, supplementing water, and autoclaving for standby;
b. liquid submerged fermentation culture: inoculating the single bacterial colony of the strain into the culture medium, performing shake culture at 100r/min, wherein the fermentation temperature is 37 ℃, the fermentation time is 36 hours, and the OD value of the fermentation liquid is 5;
c, in 2020, performing a penaeus vannamei growth promotion test on a test base of a fresh water aquatic product research institute in Zhejiang lake, Pinghu, Zhejiang, mixing lactobacillus rhamnosus fermentation liquor with feeds (purchased from Macro feeds, Inc. in Yangzhou and Penaeus vannamei) at each stage for feeding, wherein the usage amount of the fermentation liquor is 1 per mill (mL/g) of the feeds, adding the fermentation liquor in the whole process from the feeding of the feeds, and sampling and counting the average value of the length of the penaeus vannamei after 90 days;
d. the experimental results are shown in figure 1. From the figure 1, it is obvious that the shrimp body growth of the lactobacillus rhamnosus CGMCC No.20472 strain treated group is faster than that of the control group, the average value of the control group is 16.3cm, the treated group is 19.0cm and the length is increased by 16.6 percent in the third month of culture.
The above-described embodiments are only preferred embodiments of the present invention, and are not intended to limit the present invention in any way, and other variations and modifications may be made without departing from the spirit of the invention as set forth in the claims.
Sequence listing
<110> Senzo bioengineering (Huzhou) Co., Ltd
<120> Lactobacillus rhamnosus and application thereof
<130> 2020
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1433
<212> DNA
<213> Lactobacillus rhamnosus (Lactobacillus rhamnosus)
<400> 1
aatgcaagtc gaacgagttc tgattattga aaggtgcttg catcttgatt taattttgaa 60
cgagtggcgg acgggtgagt aacacgtggg taacctgccc ttaagtgggg gataacattt 120
ggaaacagat gctaataccg cataaatcca agaaccgcat ggttcttggc tgaaagatgg 180
cgtaagctat cgcttttgga tggacccgcg gcgtattagc tagttggtga ggtaacggct 240
caccaaggca atgatacgta gccgaactga gaggttgatc ggccacattg ggactgagac 300
acggcccaaa ctcctacggg aggcagcagt agggaatctt ccacaatgga cgcaagtctg 360
atggagcaac gccgcgtgag tgaagaaggc tttcgggtcg taaaactctg ttgttggaga 420
agaatggtcg gcagagtaac tgttgtcggc gtgacggtat ccaaccagaa agccacggct 480
aactacgtgc cagcagccgc ggtaatacgt aggtggcaag cgttatccgg atttattggg 540
cgtaaagcga gcgcaggcgg ttttttaagt ctgatgtgaa agccctcggc ttaaccgagg 600
aagtgcatcg gaaactggga aacttgagtg cagaagagga cagtggaact ccatgtgtag 660
cggtgaaatg cgtagatata tggaagaaca ccagtggcga aggcggctgt ctggtctgta 720
actgacgctg aggctcgaaa gcatgggtag cgaacaggat tagataccct ggtagtccat 780
gccgtaaacg atgaatgcta ggtgttggag ggtttccgcc cttcagtgcc gcagctaacg 840
cattaagcat tccgcctggg gagtacgacc gcaaggttga aactcaaagg aattgacggg 900
ggcccgcaca agcggtggag catgtggttt aattcgaagc aacgcgaaga accttaccag 960
gtcttgacat cttttgatca cctgagagat caggtttccc cttcgggggc aaaatgacag 1020
gtggtgcatg gttgtcgtca gctcgtgtcg tgagatgttg ggttaagtcc cgcaacgagc 1080
gcaaccctta tgactagttg ccagcattta gttgggcact ctagtaagac tgccggtgac 1140
aaaccggagg aaggtgggga tgacgtcaaa tcatcatgcc ccttatgacc tgggctacac 1200
acgtgctaca atggatggta caacgagttg cgagaccgcg aggtcaagct aatctcttaa 1260
agccattctc agttcggact gtaggctgca actcgcctac acgaagtcgg aatcgctagt 1320
aatcgcggat cagcacgccg cggtgaatac gttcccgggc cttgtacaca ccgcccgtca 1380
caccatgaga gtttgtaaca cccgaagccg gtggcgtaac ccttttaggg agc 1433
Claims (3)
1. Application of lactobacillus rhamnosus strain in promoting growth of aquatic animals needing bait feeding, wherein the taxonomic name of the lactobacillus rhamnosus strain is lactobacillus rhamnosus (L) ((L))Lactobacillus rhamnosus) And the strain code is as follows: LR-003, the preservation number is CGMCC number 20472, the preservation unit is China general microbiological culture Collection center (CGMCC), and the preservation unit address is as follows: the microbial research institute of the national academy of sciences No. 3, Xilu No. 1, Beijing, Chaoyang, and the preservation date is 07-30 days in 2020.
2. The use as claimed in claim 1, characterized in that a fermentation broth of lactobacillus rhamnosus is mixed with the feed for feeding, wherein the volume ml of the fermentation broth used is 1% o of the mass g of the feed.
3. The use according to claim 2, wherein the culture method of the fermentation broth of lactobacillus rhamnosus is:
(1) the formula of the culture medium is as follows: 1% of sucrose mass concentration, 1% of yeast powder mass concentration and 0.5% of sodium chloride mass concentration, supplementing water, and sterilizing for later use;
(2) liquid submerged fermentation culture: and (3) inoculating the single bacterial colony of the strain into the culture medium, performing shaking culture at 120 r/min, wherein the fermentation temperature is 35 ℃, the fermentation time is 36 hours, and the OD value of the fermentation liquid is 6.
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| CN110628663B (en) * | 2019-08-02 | 2021-12-17 | 华南理工大学 | Lactobacillus rhamnosus and high-density culture method and application thereof |
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