CN115044514A

CN115044514A - Lactobacillus plantarum, pet health product and using method thereof

Info

Publication number: CN115044514A
Application number: CN202210773062.0A
Authority: CN
Inventors: 韩冰; 王金全; 陶慧; 王振龙; 王秀敏; 刘杰
Original assignee: Feed Research Institute of Chinese Academy of Agricultural Sciences
Current assignee: Feed Research Institute of Chinese Academy of Agricultural Sciences
Priority date: 2022-06-30
Filing date: 2022-06-30
Publication date: 2022-09-13

Abstract

The invention belongs to the field of microorganisms and pet health products, and provides lactobacillus plantarum, a pet health product and a using method thereof. The preservation number of the Lactobacillus plantarum (Lactobacillus plantarum) is CGMCC No. 24558. The pet health product comprises the bacterial powder of the lactobacillus plantarum. The product of the invention has good effects of reducing the content of helicobacter pylori and increasing the quantity of bifidobacteria in intestinal tracts for cats and dogs, and can improve the intestinal health of pets. The pet health care product which is prepared by matching the feed additive with the specific components has a synergistic effect, can assist in treating intestinal diseases of pets, reduce the number of helicobacter pylori in the bodies of the pets, increase the number of beneficial bacteria in the intestinal tracts, remarkably improve the intestinal health of the pets, reduce diarrhea, promote fat digestion and utilization, regulate fat metabolism function and improve the digestive function of the pets.

Description

Lactobacillus plantarum, pet health product and using method thereof

Technical Field

The invention relates to the field of microorganisms and pet health products, in particular to lactobacillus plantarum, a pet health product and a using method thereof.

Background

With the improvement of living standard in recent years, raising pets has become a common living mode for relieving mental stress and promoting physical and mental health of people, and pet owners pay more and more attention to pet health.

Helicobacter pylori is a common bacterium in the digestive tract of pets and humans. The bacterium can survive well under the acidic condition of the stomach. In human, however, helicobacter pylori may cause chronic gastritis and even gastric cancer in severe cases, and it is recognized that helicobacter pylori is a high risk factor for gastric cancer. In pets, discomfort symptoms such as gastrointestinal tract and the like appear when helicobacter pylori infection occurs, but generally, the helicobacter pylori infection cannot easily cause diseases, and the current mechanism is not clear. However, H.pylori can be transmitted by oral-oral, oral-faecal, human-pet transmission routes, and the pet owner is at risk of contracting H.pylori.

At present, the treatment of H.pylori in humans is mostly completely treated by triple or quadruple antibiotic therapy, but there is no clear treatment regimen for pets. Therefore, the invention provides a scheme which can play a role in assisting in treating helicobacter pylori of pets, and plays an extremely important role in improving the intestinal health of pets.

Disclosure of Invention

Aiming at the problems in the prior art, the invention provides lactobacillus plantarum, a pet health product and a using method thereof.

In a first aspect, the invention provides a strain of Lactobacillus plantarum (Lactobacillus plantarum), which is preserved in the common microorganism center of the China committee for culture Collection of microorganisms with the preservation date of 2022 years, 3 months and 21 days, and the preservation number of the strain is CGMCC No. 24558. The preservation address is as follows: the institute of microbiology, national academy of sciences, No. 3, West Lu No.1, Beijing, Chaoyang, Beicheng, area, Beichen.

According to the lactobacillus plantarum provided by the invention, the nucleotide sequence of 16SrDNA of the lactobacillus plantarum is as follows: SEQ ID NO. 1.

According to the lactobacillus plantarum provided by the invention, the lactobacillus plantarum is obtained by separating and purifying the excrement of cats.

In a second aspect, the invention provides the use of the lactobacillus plantarum described above for improving the intestinal health of cat and dog pets.

The lactobacillus plantarum provided by the invention has a good effect of reducing the content of helicobacter pylori on pets such as cats and dogs, can improve the intestinal health of pets, and has a good application prospect.

In a third aspect, the invention provides a pet health product, which comprises the lactobacillus plantarum bacterial powder.

According to the pet health product provided by the invention, the viable bacteria content of the lactobacillus plantarum in the pet health product is not less than 10 ⁸ CFU/g。

According to the pet health product provided by the invention, the preparation method of the lactobacillus plantarum bacterium powder comprises the following steps:

inoculating lactobacillus plantarum into a 200mLMRS culture medium, and culturing at 37 ℃ for 24h to prepare a seed solution; inoculating the seed solution into 1L culture medium according to the inoculation amount of 2%, culturing for 48h, centrifuging the culture solution at 10000rpm for 5min, washing the precipitate with sterile water for three times, and making into bacterial powder by vacuum freeze-drying with skimmed milk powder, galactose, trehalose, etc. as protective agent.

According to the pet health product provided by the invention, the pet health product further comprises bile acids, vitamin D and plant extracts.

According to the invention, the research discovers that the synergistic effect can be generated by compounding the bile acid substances with the lactobacillus plantarum powder, the vitamin D and the plant extract, and the obtained pet health-care product can obviously reduce the content of helicobacter pylori in excrement, improve the content of probiotics such as bifidobacterium and the like in the excrement and improve the intestinal health of pets. Moreover, the obtained pet health product can obviously improve the digestion utilization rate of fat, regulate fat metabolism, obviously reduce the content of cholesterol in blood, improve the contents of amylase, alanine transferase and aspartate transferase, well improve the fat metabolism function of pets and promote the health of pets.

It should be noted that, even though the name of the product is not pet health products (such as feed additive, helicobacter pylori inhibitor, etc.), the product is not the pet health product of the present invention as long as the effective components of the product comprise lactobacillus plantarum powder, bile acids, vitamin D and plant extract.

According to the pet health product provided by the invention, the mass ratio of the lactobacillus plantarum powder to the bile acids to the vitamin D to the plant extract is 1-6:0.001-3:0.01-0.2: 0.01-1.

According to the pet health product provided by the invention, the bile acid is one or more of allocholic acid, hyocholic acid, hyodeoxycholic acid or chenodeoxycholic acid.

According to the pet health product provided by the invention, the plant extract comprises one or more of turmeric extract, licorice extract, green tea extract and mushroom extract.

According to the pet health product provided by the invention, the pet health product is used for pets such as cats and dogs.

According to the pet health product provided by the invention, the dosage form of the pet health product is tablet or powder. When the powder is prepared, the preparation method comprises mixing the raw materials in proportion; when the tablet is prepared, the raw materials are mixed and then are pressed by a tablet machine.

In a fourth aspect, the invention also provides a using method of the pet health product.

The using method is that the pet health care product is evenly mixed in pet feed or water with the dosage of 100-500mg per day, or directly orally taken by the pet for 21-42 days.

Further preferably, the pet health product is uniformly mixed in the pet feed or water in an amount of 200-300mg per day, or is directly orally taken by the pet for 28-35 days.

It should be noted that the raw materials involved in the present invention are all materials allowed to be used in the feed additive list, and the lactobacillus plantarum comes from pet sources and can be directly used for pets without generating exclusivity.

The invention provides lactobacillus plantarum, a pet health product and a using method thereof. The lactobacillus plantarum has a good effect of reducing the content of helicobacter pylori on cats and dogs, and can improve the intestinal health of pets. The pet health care product which is prepared by matching the feed additive with the specific components has a synergistic effect, can assist in treating intestinal diseases of pets, reduce the number of helicobacter pylori in the bodies of the pets, remarkably improve the health of the intestinal tracts of the pets, reduce the diarrhea condition, promote the digestion and utilization of fat, regulate the fat metabolism function and improve the digestion function of the pets.

Drawings

FIG. 1 shows the results of intestinal microbial diversity at day 28 of different treatments (blank control, example 5 sample group) (the legend on the right corresponds to the different regions of the bar graph on the left from top to bottom);

FIG. 2 shows the results of the abundance of helicobacter at day 28 for the different treatments (blank control, example 5 sample);

fig. 3 shows the results of bifidobacterium abundance at day 28 for different treatments (blank control, example 5 sample).

Detailed Description

In order to make the objects, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention are clearly and completely described below, and it is obvious that the described embodiments are a part of the embodiments of the present invention, but not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

The starting materials used in the present invention are commercially available unless otherwise specified.

Example 1 isolation and identification of Strain L11

Mixing 10g of feces of healthy cats with 100mL of sterile MRS liquid culture medium, standing at 37 ℃ for 48h, transferring a small amount of liquid culture to an MRS solid culture medium for culture, performing separation and purification for multiple times to obtain a single colony, and naming the obtained single colony as L11.

Inoculating a single L11 colony into a liquid culture medium, culturing for 24h, extracting the genome, using the genome as a template, performing PCR amplification by using a universal primer of 16SrDNA, sequencing the sequence obtained after amplification, and performing BLAST comparison on the sequence in an NCBI database, wherein the result shows that the strain L11 has higher homology with lactobacillus plantarum, the nucleotide sequence of the 16SrDNA is shown as SEQ ID NO.1, and the preservation number is CGMCC No. 24558.

Example 2 characterization of Strain L11

Escherichia coli is selected as an indicator bacterium, and an antibacterial test is carried out by using L11 fermentation liquor (L11 of lactobacillus plantarum is inoculated into a 200mLMRS culture medium and cultured for 24 hours at 37 ℃ to prepare seed liquid, and the seed liquid is inoculated into 1L culture medium according to the inoculum concentration of 2% and cultured for 48 hours to obtain L11 fermentation liquor). Bacteriostatic tests show that the strain has good bacteriostatic effect on escherichia coli. The acid production performance of the bacterium is high, and through HPLC analysis, the fermentation liquor of the bacterium contains high-content small molecular organic acids such as acetic acid and lactic acid, and the content of the generated lactic acid is up to 24.0 g/L.

Example 3 preparation of Pet health products

The embodiment provides a pet health product, and the preparation method thereof comprises the following steps:

centrifuging the L11 fermentation liquid at 10000rpm for 5min, washing the precipitate with sterile water for three times, using skimmed milk powder, galactose and trehalose as protective agents, and freeze-drying in a low-temperature vacuum freeze dryer to obtain bacterial powder for later use.

Mixing the obtained bacterial powder with hyocholic acid, a licorice extract and vitamin D according to a mass ratio of 1: 0.01: 0.02: 0.01, and making into tablet or powder to obtain pet health product.

Example 4

Mixing the obtained bacterial powder with chenodeoxycholic acid, turmeric extract, liquorice extract and vitamin D according to the mass ratio of 1: 0.02: 0.06: 0.01: 0.01, and making into tablet or powder to obtain pet health product.

Example 5

Mixing the obtained bacterial powder with allocholic acid, a mushroom extract and vitamin D according to a mass ratio of 1: 0.005: 0.2: 0.05, and making into tablet or powder to obtain pet health product.

Example 6

Mixing the obtained bacterial powder with hyodeoxycholic acid, green tea extract and vitamin D according to the mass ratio of 1: 0.06: 0.8: 0.02 mixing, and making into tablet or powder for pet health product.

Comparative example 1

This comparative example provides a pet health product which differs from example 3 in that the same amount (in terms of viable count) of raw powder of lactobacillus plantarum obtained in CN112940983A example 4 was used in place of the powder of lactobacillus plantarum of the present invention.

Comparative example 2

This comparative example provides a pet care product which differs from example 3 in that the same amount (in terms of viable count) of the helicobacter pylori inhibitor obtained in CN113980878A example 8 was used in place of the lactobacillus plantarum powder used in the present invention.

Comparative example 3

The comparative example provides a pet health product, which comprises lactobacillus plantarum powder, a green tea extract, a licorice extract and vitamin D in example 3 of the invention, and the mass ratio of the lactobacillus plantarum powder to the green tea extract to the licorice extract to the vitamin D is 1: 0.1: 0.05: 0.01.

pet experimental verification

Selecting 48 adult cats with similar ages and healthy bodies, dividing the adult cats into 8 groups (6 cats in each group) for pet test verification, freely eating and drinking water during feeding, feeding samples according to the dosage required by the adult cats, and adding the pet health care products (respectively products of examples 3, 4, 5 and 6 and comparative examples 1, 2 and 3) into the feeds of the other groups except the blank control group to ensure that the amount of the pet health care products taken by each pet per day is 100-500 mg.

Tests for body weight, fat digestibility, cholesterol and intestinal microbial diversity were performed on days 0 and 28, respectively, and the results of the tests are shown in tables 1-2 and fig. 1-3.

TABLE 1 Effect of Pet health products on adult Cat growth index

TABLE 2 Effect of Pet health products on adult Cat blood Biochemical indicators

As can be seen from tables 1 and 2, the use of the pet health product of the present invention is more beneficial to the reduction of fat, the control of body weight, and the prevention of cholesterol increase in pets.

As can be seen from FIGS. 1 to 3, the use of the pet health product of the present invention can reduce the abundance of helicobacter (harmful bacteria) and increase the abundance of bifidobacterium (beneficial bacteria).

Finally, it should be noted that: the above examples are only intended to illustrate the technical solution of the present invention, but not to limit it; although the present invention has been described in detail with reference to the foregoing embodiments, it will be understood by those of ordinary skill in the art that: the technical solutions described in the foregoing embodiments may still be modified, or some technical features may be equivalently replaced; and such modifications or substitutions do not depart from the spirit and scope of the corresponding technical solutions of the embodiments of the present invention.

Sequence listing

<110> institute of feed of Chinese academy of agricultural sciences

<120> Lactobacillus plantarum, pet health product and use method thereof

<130> KHP221113725.4

<160> 1

<170> SIPOSequenceListing 1.0

<210> 1

<211> 1480

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 1

accttggggg ggtgccctat acatgcaagt cgaacgaact ctggtattga ttggtgcttg 60

catcatgatt tacatttgag tgagtggcga actggtgagt aacacgtggg aaacctgccc 120

agaagcgggg gataacacct ggaaacagat gctaataccg cataacaact tggaccgcat 180

ggtccgagct tgaaagatgg cttcggctat cacttttgga tggtcccgcg gcgtattagc 240

tagatggtgg ggtaacggct caccatggca atgatacgta gccgacctga gagggtaatc 300

ggccacattg ggactgagac acggcccaaa ctcctacggg aggcagcagt agggaatctt 360

ccacaatgga cgaaagtctg atggagcaac gccgcgtgag tgaagaaggg tttcggctcg 420

taaaactctg ttgttaaaga agaacatatc tgagagtaac tgttcaggta ttgacggtat 480

ttaaccagaa agccacggct aactacgtgc cagcagccgc ggtaatacgt aggtggcaag 540

cgttgtccgg atttattggg cgtaaagcga gcgcaggcgg ttttttaagt ctgatgtgaa 600

agccttcggc tcaaccgaag aagtgcatcg gaaactggga aacttgagtg cagaagagga 660

cagtggaact ccatgtgtag cggtgaaatg cgtagatata tggaagaaca ccagtggcga 720

aggcggctgt ctggtctgta actgacgctg aggctcgaaa gtatgggtag caaacaggat 780

tagataccct ggtagtccat accgtaaacg atgaatgcta agtgttggag ggtttccgcc 840

cttcagtgct gcagctaacg cattaagcat tccgcctggg gagtacggcc gcaaggctga 900

aactcaaagg aattgacggg ggcccgcaca agcggtggag catgtggttt aattcgaagc 960

tacgcgaaga accttaccag gtcttgacat actatgcaaa tctaagagat tagacgttcc 1020

cttcggggac atggatacag gtggtgcatg gttgtcgtca gctcgtgtcg tgagatgttg 1080

ggttaagtcc cgcaacgagc gcaaccctta ttatcagttg ccagcattaa gttgggcact 1140

ctggtgagac tgccggtgac aaaccggagg aaggtgggga tgacgtcaaa tcatcatgcc 1200

ccttatgacc tgggctacac acgtgctaca atggatggta caacgagttg cgaactcgcg 1260

agagtaagct aatctcttaa agccattctc agttcggatt gtaggctgca actcgcctac 1320

atgaagtcgg aatcgctagt aatcgcggat cagcatgccg cggtgaatac gttcccgggc 1380

cttgtacaca ccgcccgtca caccatgaga gtttgtaaca cccaaagtcg gtggggtaac 1440

cttttaggaa ccagccgcct aaggttgaca gaaattgccc 1480

Claims

1. The Lactobacillus plantarum is characterized in that the preservation number of the Lactobacillus plantarum (Lactobacillus plantarum) is CGMCC No. 24558.

2. The lactobacillus plantarum of claim 1, wherein the lactobacillus plantarum has the nucleotide sequence shown in SEQ ID No.1 for 16 SrDNA.

3. Use of lactobacillus plantarum as defined in claim 1 or 2 for improving gut health in cats and dogs pets.

4. A pet health product comprising the powder of lactobacillus plantarum of claim 1 or 2.

5. The pet health product of claim 4, wherein the viable bacteria content of Lactobacillus plantarum in the pet health product is not less than 10 ⁸ CFU/g。

6. The pet food of claim 4, further comprising bile acids, vitamin D, and plant extracts.

7. The pet health product of claim 6, wherein the mass ratio of the Lactobacillus plantarum powder, the bile acids, the vitamin D and the plant extract is 1-6:0.001-3:0.01-0.2: 0.01-1.

8. The pet health product of claim 7, wherein the bile acid is one or more of allocholic acid, hyocholic acid, hyodeoxycholic acid or chenodeoxycholic acid;

and/or the plant extract comprises one or more of curcuma extract, liquorice extract, green tea extract and mushroom extract.

9. The pet health product of any one of claims 4-8, wherein the pet health product is for a cat, a dog pet;

and/or the dosage form of the pet health product is a tablet or powder.

10. The method for using a pet health product according to any of claims 4-9, wherein the pet health product is mixed in the pet food or water in an amount of 100 mg/day or orally administered directly to the pet for 21-42 days continuously.