CN116369427A - Application of lactobacillus johnsonii in preparation of antioxidant and immunity-improving products - Google Patents
Application of lactobacillus johnsonii in preparation of antioxidant and immunity-improving products Download PDFInfo
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- CN116369427A CN116369427A CN202211738422.XA CN202211738422A CN116369427A CN 116369427 A CN116369427 A CN 116369427A CN 202211738422 A CN202211738422 A CN 202211738422A CN 116369427 A CN116369427 A CN 116369427A
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- lactobacillus johnsonii
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- lactobacillus
- immunity
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- 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
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- 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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- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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- A23K50/00—Feeding-stuffs specially adapted for particular animals
- A23K50/30—Feeding-stuffs specially adapted for particular animals for swines
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- 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 discloses an application of lactobacillus johnsonii in preparing antioxidant and immunity-improving products, wherein the products are microbial agents or animal feeds, and the viable count of the lactobacillus johnsonii in the animal feeds is 1 multiplied by 10 7 ～2.5×10 7 CFU/g, wherein the animal feed is piglet feed; the lactobacillus johnsonii is classified and named as Lactobacillus johnsonii RS-7, and is registered and preserved in China Center for Type Culture Collection (CCTCC) on the 29 th year 2022, wherein the preservation number is as follows: m20221532. The lactobacillus johnsonii bacterial suspension and the cell extract of the invention have certain functionsDPPH free radical, superoxide anion free radical and hydroxyl free radical scavenging ability and certain reducing ability; the lactobacillus johnsonii can be applied to preparation of products for resisting oxidization and improving immunity.
Description
Technical Field
The invention relates to the technical field of microorganisms, in particular to application of lactobacillus johnsonii in preparation of products for resisting oxidization and improving immunity.
Background
Along with the expansion of the current pig raising scale, in order to reduce the raising risk and increase the raising income, a better growing environment is provided for the piglets, and the immunity and the growing performance of the piglets are improved. The growth speed of the piglet in the early stage is high, and the piglet is a key stage of functional development of various organs and determines the growth and production performance in the later stage. During the period, the quantity and diversity of the intestinal bacteria of the pigs are greatly improved, and succession and change of the intestinal bacteria are closely related to intestinal development, health level, digestion and immune function of the pigs. When a piglet weans, the transformation of food types and stress response initiated by the transformation can cause the decline of intestinal health function and the obvious change of intestinal flora composition of the piglet within a certain period of time. During the period, the diarrhea rate of the piglets is obviously increased, the growth performance is reduced, the number and proportion of probiotics such as lactobacillus are greatly reduced, and the health and growth of the pigs are affected. Improving and maintaining intestinal health, and promoting the stability and balance of intestinal microecological systems is one of key factors for guaranteeing early rapid growth of piglets. In order to maintain animal health and reduce the adverse effects of weaning without antibiotics, additives such as probiotics are often used in animal production to stimulate the intestinal flora to achieve optimal microecological balance. The original bacteria separated from the animal can better adapt to the gastrointestinal tract environment of the same kind of animals, so that the probiotic property of the animals can be better exerted, and the maximized treatment and health care effects are obtained; lactobacillus johnsonii (Lactobacillus johnsonii, L.johnsonii) is classified in the genus Lactobacillus. Studies show that lactobacillus johnsonii has good probiotics, such as promoting the growth of piglets and reducing diarrhea of piglets, but few reports of lactobacillus johnsonii of swine origin exist in the prior art, and no related reports of antioxidation and immunity improvement of lactobacillus johnsonii of swine origin exist.
Disclosure of Invention
It is an object of the present invention to address at least the above problems and/or disadvantages and to provide at least the advantages described below.
To achieve these objects and other advantages and in accordance with the purpose of the invention, there is provided a use of lactobacillus johnsonii for preparing an antioxidant and immunity enhancing product, which is a microbial agent or an animal feed.
Preferably, the viable count of Lactobacillus johnsonii in the animal feed is 1×10 7 ～2.5×10 7 CFU/g。
Preferably, the animal feed is a piglet feed.
Preferably, the bacterial suspension and the cell extract of lactobacillus johnsonii have certain DPPH free radical, superoxide anion free radical and hydroxyl radical scavenging capacity and certain reducing capacity.
Preferably, the lactobacillus johnsonii is classified and named as Lactobacillus johnsonii RS-7, and is registered and preserved in China Center for Type Culture Collection (CCTCC) on the year 09 and 29 of 2022, wherein the preservation number is as follows: m20221532.
Preferably, the lactobacillus johnsonii is derived from fresh pig manure.
Preferably, the sequence of the 16S rDNA of lactobacillus johnsonii is shown as SEQ ID NO. 1.
The invention at least comprises the following beneficial effects: the lactobacillus johnsonii bacterial suspension and the cell extract of the invention have certain DPPH free radical, superoxide anion free radical, hydroxyl radical scavenging capability and certain reducing capability; the lactobacillus johnsonii can be applied to preparation of products for resisting oxidization and improving immunity.
Additional advantages, objects, and features of the invention will be set forth in part in the description which follows and in part will become apparent to those having ordinary skill in the art upon examination of the following or may be learned from practice of the invention.
Description of the drawings:
FIG. 1 is a diagram showing the effect of the inhibition zones of RS-3, RS-6 and RS-7 of the invention, wherein, E.coli A; salmonella B; staphylococcus aureus; 3: RS-3;6: RS-6;7: RS-7;
FIG. 2 is a colony morphology of Lactobacillus johnsonii RS-7 of the present invention;
FIG. 3 is a gram stain (. Times.1000) of Lactobacillus johnsonii RS-7 of the present invention;
FIG. 4 is a genetic evolutionary tree of Lactobacillus johnsonii RS-7 of the present invention;
FIG. 5 is an in vitro antioxidant capacity profile of Lactobacillus johnsonii RS-7 of the present invention;
FIG. 6 is a graph showing the effect of Lactobacillus johnsonii RS-7 of the present invention on the antioxidant capacity of piglets;
FIG. 7 is a graph showing the effect of Lactobacillus johnsonii RS-7 of the present invention on the antioxidant capacity of piglets;
FIG. 8 shows the growth curve of lactobacillus suis RS-7 of the present invention.
The specific embodiment is as follows:
the present invention is described in further detail below with reference to the drawings to enable those skilled in the art to practice the invention by referring to the description.
It will be understood that terms, such as "having," "including," and "comprising," as used herein, do not preclude the presence or addition of one or more other elements or groups thereof.
Example 1:
use of lactobacillus johnsonii for the preparation of an antioxidant and immunity enhancing product which is a microbial agent or an animal feed;
the lactobacillus johnsonii is classified and named as Lactobacillus johnsonii RS-7, and is registered and preserved in China Center for Type Culture Collection (CCTCC) on the 29 th year 2022, wherein the preservation number is as follows: m20221532;
lactobacillus johnsonii screening and isolation:
taking fresh pig manure of a Sichuan Mianyang local pig as a sample, aseptically weighing 0.1g of pig manure, carrying out gradient dilution by using PBS solution, uniformly coating the diluted solution on an MRS solid culture medium containing calcium carbonate, carrying out constant-temperature culture at 37 ℃ for 24 hours, picking a single bacterial colony with a calcium-dissolving ring, streaking and inoculating the single bacterial colony into the MRS solid culture medium, carrying out constant-temperature culture at 37 ℃ for 24 hours, and purifying for 3 times until a pure bacterial strain is obtained; primarily screening and separating 29 gram-positive lactobacillus with the serial numbers of RS-1 to RS-29;
screening characteristics of swine lactobacillus antagonistic antigen bacteria:
after the MRS broth solid culture medium is sterilized and cooled to about 50 ℃, pouring the broth solid culture medium into a sterile culture dish, after the flat plate is naturally aired, respectively transferring 0.1mL of indicator bacteria liquid (108 CFU/mL) to uniformly coat on the flat plate, after the flat plate is naturally aired, punching holes on the flat plate, adding 0.1mL of bacteria liquid to be tested into the holes, standing, and after the bacteria liquid permeates the flat plate, culturing at a constant temperature of 37 ℃ for 24 hours, and measuring the size of a bacteriostasis zone. The indicator bacteria are escherichia coli, staphylococcus aureus and salmonella respectively; by inhibiting Escherichia coli, staphylococcus aureus and salmonella, lactobacillus 3 strain with certain antibacterial effect, RS-3, RS-6 and RS-7 are obtained by preliminary screening, wherein the antibacterial effect of RS-7 is optimal, and the antibacterial results are shown in table 1 and figure 1. The dominant strain passing the bacteriostasis test is preserved.
TABLE 1
Identification of lactobacillus suis RS-7:
morphological identification:
in a sterile operation table, the purified lactobacillus RS-7 is streaked on an MRS plate, and is respectively cultured for 24 hours at 37 ℃, and the characteristics of the size, shape, glossiness, color, transparency degree and the like of a single colony are observed and recorded.
All isolates were gram stained, staining procedure referenced to staining kit, and bacterial status was observed and recorded under microscope.
Colony morphology of each strain is shown in fig. 2 and table 2;
gram staining observations revealed that the screened lactic acid bacteria were rod-shaped, as shown in figure 3.
TABLE 2
And (3) identifying physiological and biochemical characteristics:
the physiological and biochemical characteristics of the RS-7 strain are detected according to the steps in the biochemical detection kit, and the physiological and biochemical identification results are shown in the table 3;
control analysis was performed with reference to "Manual of identification of common bacterial System" and "Manual of identification of Bojie's bacteria", and RS-7 was primarily judged to be Lactobacillus-producing bacteria.
TABLE 3 Table 3
| Project | Results | Project | Results |
| Catalase enzyme | - | Sorbitol | - |
| VP reaction | - | Radix Schefflerae Arboricolae | - |
| Methyl red reaction | - | Glucose | - |
| Starch hydrolysis | - | Raffinose | - |
| Cellobiose | - | Sucrose | - |
| Maltose | - | Gelatin | + |
| Mannitol (mannitol) | - |
Positive is indicated by +; -negative representation
Molecular biology identification:
PCR amplification was performed using bacterial identification 16S rDNA universal primers 27F and 1492R, the primer sequences are shown in Table 4, and the primers were synthesized by Shanghai Biotechnology Co., ltd;
TABLE 4 Table 4
The PCR amplified template was activated fresh bacterial liquid, 50. Mu.L in system, as detailed in Table 5.
TABLE 5
| Composition of the components | Volume (mu L) |
| 10×PCR Buffer | 5.0 |
| dNTP | 3.0 |
| Template | 1.0 |
| 27F | 0.5 |
| 1492R | 0.5 |
| rTaq DNase | 0.3 |
| ddH 2 O | 39.7 |
The PCR amplification procedure was as follows: 95 ℃ for 5min; 45s at 95 ℃; 45s at 55 ℃;72 ℃ for 1min; repeat 35 cycles; and at 72℃for 8min. Sequencing the PCR amplified products after 1% agarose gel electrophoresis identification in Shanghai Biotechnology, and performing BLAST comparison analysis in GenBank; mapping the genetic evolutionary tree, the result shows that RS-7 is on the same branch as the reference strain L.johnsonii (ATCC 33200), and the relatedness is nearest (FIG. 4), so that the RS-7 can be judged to be lactobacillus johnsonii; and sequencing to obtain the 16S rDNA sequence of the lactobacillus johnsonii Lactobacillus johnsonii RS-7, wherein the sequence is shown in SEQ ID NO. 1;
biological characterization of lactobacillus swine RS-7:
after the strain is activated for one generation, inoculating 100mL of MRS liquid culture medium according to the inoculum size of 2%, shake culturing at 37 ℃, taking 5mL of strain liquid to be measured every 4 hours, measuring the concentration (OD value) of the strain liquid at 600nm wavelength by using an ultraviolet spectrophotometer, recording for 24 hours in total, and as shown in figure 8, determining the growth curve of lactobacillus suis RS-7, wherein RS-7 can be found to enter the logarithmic phase after 4 hours, and enter the stationary phase after 8 hours.
Example 2:
use of lactobacillus johnsonii for the preparation of an antioxidant and immunity enhancing product:
the articles of manufacture of the present application are useful for enhancing the antioxidant capacity and immunity of an animal, thereby reducing the induction of disease in the animal. The lactobacillus johnsonii application data are as follows:
in vitro antioxidant activity of lactobacillus zoonotis of the present application:
1. preparation of bacterial liquid and in-vitro antioxidation detection
1. Reagent(s)
Absolute ethyl alcohol, potassium ferricyanide, PBS buffer solution, acetic acid, ferric trichloride, ascorbic acid, hydrogen peroxide, ferrous sulfate, 3, 5-dinitrosalicylic acid (DNS), pyrogallic acid (pyrogallol), a trihydroxymethyl aminomethane-hydrochloric acid buffer solution (Tris), a phosphate buffer solution, an o-phenanthroline solution and the like, which are all analytically pure.
2. Extraction of sample bacterial liquid
Reference Gao Lie [ Gao Lie ] research on diversity and antioxidant properties of lactic acid bacteria in conventional fermented yak milk in Qinghai-Tibet plateau [ D ]]University of Lanzhou 2020.DOI:10.27204/d.cnki.glzhu.2020.002996.]Different components of the strain are prepared by the method of (a). The strain was inoculated in MRS liquid medium, cultured at 37℃for 24 hours, centrifuged at 8000g for 10min at 4℃and the supernatant and the cell were collected, respectively, and the supernatant was Fermentation Supernatant (FS). The obtained thalli are washed 2 to 3 times by sterile water, resuspended in PBS and adjusted to have the density of 1 multiplied by 10 9 CFU/mL, divided equally into 2 parts, part 1 as cell (IC); in the other 1 part, performing ultrasonic disruption under ice bath condition (250W, ultrasonic treatment for 5s, interval of 10s, total 10 min), centrifuging for 10min at 8000g, and collecting supernatant to obtain cell-free extract (CFE);
3. in vitro antioxidant Activity assay
3.1 measurement of DPPH radical scavenging ability
1mL of a sample is taken and placed in a test tube, 2mL (the concentration is 0.2 mmol/L) of DPPH absolute ethanol solution is added, after uniform mixing, the mixture is subjected to light-proof reaction at room temperature for 30min, at 4 ℃,8000g, centrifugation is carried out for 10min, the supernatant is taken, the absorbance is measured at 517nm, and deionized water is used for zeroing. Each group was averaged over 3 replicates.
DPPH clearance (%) = [1- (a sample-a blank)/a control ] ×100%
( Blank group: DPPH was replaced with equal volume of absolute ethanol; control group: the sample solution is replaced by equal volume distilled water, and the blank zeroing is carried out by equal volume distilled water and absolute ethyl alcohol mixed solution )
Liquid configuration: 0.2mmol/L DPPH absolute ethanol solution: 50mg DPPH is weighed, 634mL of absolute ethyl alcohol is added, and the mixture is shaken well.
3.2 determination of the hydroxy radical (.OH) scavenging Activity (Fenton method)
The Fenton method uses H 2 O 2 And Fe (Fe) 2+ The Fenton reaction takes place by mixing. While producing OH with very high reactivity, salicylic acid can capture OH and produce a colored substance (dihydroxybenzoic acid) that has a maximum absorption at 510 nm. But if addWhen a certain amount of substance with scavenging ability is added, competition with salicylic acid is generated, so that the generation amount of colored substance (dihydroxybenzoic acid) is reduced, the absorbance of the solution is changed to a certain extent, and the activity of the antioxidant can be measured according to the change of absorbance.
H 2 O 2 +Fe 2+ →·OH+OH - +Fe 3+
Liquid configuration: (1) FeSO 4 ·7H 2 O: precisely weighing 0.05g of ferrous sulfate heptahydrate, placing in a 100ml volumetric flask, adding distilled water to scale, and shaking to obtain 1.8nmol/L ferrous sulfate solution.
(2) Salicylic acid-ethanol: salicylic acid with the precise weighing amount of 0.0249g is placed in a 100ml volumetric flask, absolute ethyl alcohol is added to the scale, and after shaking evenly and stabilization, 1.8nmol/L salicylic acid-ethanol solution is obtained.
③0.3％H 2 O 2 Solution: precisely measuring 1.0ml 30% hydrogen peroxide, placing in 100ml volumetric flask, adding distilled water to scale, and mixing to obtain 0.3% H 2 O 2 A solution.
The method comprises the following specific steps: 2mL of FeSO 1.8mmoL/mL was added sequentially to the tube 4 1mL of sample and 0.3% H 2 O 2 0.1mL of solution is uniformly shaken, 1.5mL of salicylic acid-ethanol solution with the concentration of 1.8mmol/mL is added, the solution is uniformly shaken, and after standing for 30min at 30 ℃, the absorbance value Ai of the sample group is measured at 510 nm; h in the sample group 2 O 2 The solution is changed into distilled water with equal quantity, and the reaction is carried out, and the distilled water is taken as a sample reference group and the absorbance value Aj is measured; the samples in the sample group were replaced with distilled water in equal amounts, and the reaction was performed, and the absorbance Ao was measured as a blank group. The hydroxyl radical scavenging rate was calculated according to the following formula:
clearance (%) =ao- (Ai-Aj)/ao×100%
3.3 superoxide anion radical (O) 2 - Determination of scavenging Activity (pyrogallol method)
Pyrogallol is extremely susceptible to autoxidation in alkaline environments, yielding colored intermediates and superoxide anions O 2 - ·，O 2 - And has a catalytic action on the autoxidation reaction, and determines O based on the amount of colored substances generated 2 - The amount of formation. When the antioxidant active substance is added, the autoxidation of the pyrogallol can be weakened, so that the reduction trend of the colored substances is utilized to judge the antioxidant activity.
Liquid configuration: (1) 25nmol/L of pyrogallol: accurately weighing 0.315g of pyrogallol, placing in a 100ml volumetric flask, adding 10ml of 0.1mol/L hydrochloric acid, adding distilled water to scale, and shaking to obtain 25nmol/L pyrogallol. The pyrogallol is prepared and used at present, and reacts at 20 ℃.
(2) Tris-HCl: 3.0285g of Tris reagent is precisely weighed and placed in a 500ml volumetric flask, 114.5ml of 0.1mol/L hydrochloric acid is added, finally distilled water is added to the scale, and shaking is carried out, thus obtaining 0.05mol/L buffer solution.
The method comprises the following specific steps: taking out a test tube, adding 7.5ml of Tris-HCl buffer solution, placing the test tube in a water bath kettle at 25 ℃ for preheating for 20min, respectively adding 1ml of sample solution and 0.5ml of 25mmol/L pyrogallol solution, mixing, placing the mixed solution in the water bath kettle at 25 ℃ for reacting for 5min, adding 1ml of concentrated HCl for terminating the reaction, and measuring the absorbance Ai of the solution at 320nm wavelength under an ultraviolet-visible spectrophotometer as a sample group; the sample reference group replaces the pyrogallol solution with the same volume of distilled water and the absorbance Aj of the solution is determined; the absorbance Ao of the solution was measured with the same volume of distilled water instead of the sample solution in the blank group. O is calculated according to the following formula 2 - Clearance rate:
clearance (%) =ao- (Ai-Aj)/ao×100%
3.4 determination of the reducing Capacity
Liquid configuration: (1) phosphate buffer solution of 0.2mol/L, pH 6.6.6: first preparing first solution 0.2MNA 2 HPO 4 (17.19g Na 2 HPO 4 ·12H 2 O+240mL H 2 O) and ethyl acetate 0.2M NaH 2 PO 4 (11.23gNaH 2 PO 4 ·2H 2 O+360mL H 2 O); 40mL of solution A and 60mL of solution B are taken, and the mixed solution is the solution with the pH of 6.6.
(2) 1% potassium ferricyanide: 1g potassium ferricyanide+100 mL;
(3) trichloroacetic acid 1mL+9mL H 2 O；
(4) 0.1% ferric trichloride: 0.1g of ferric trichloride+100 mL.
A sample of 0.5mL was placed in a test tube, 0.5mL of phosphate buffer solution of 0.2mol/L, pH 6.6.6 was added, and then 0.5mL of 1% potassium ferricyanide was added, followed by rapid ice-bath cooling after 20min at 50 ℃. Then, 10% trichloroacetic acid (0.5 mL) was added, the mixture was centrifuged at 4000r/min for 10min, 1mL of the supernatant was obtained, and 1mL of distilled water and 1mL of 0.1% ferric trichloride were added and mixed uniformly. The mixture was left at room temperature for 10 minutes, and the absorbance was measured at 700 nm. Each sample was replicated 3 times. The larger the absorbance, the stronger the reducing power of the sample to be measured. Each group was averaged over 3 replicates.
The bacterial suspension (IC) and the cell extract (CFE) of the lactobacillus RS-7 have certain DPPH free radical, superoxide anion free radical, hydroxyl free radical scavenging capability and certain reducing capability, which shows that the strain has certain antioxidant activity in vitro. The results are shown in Table 6 and FIG. 5 (wherein, (A) DPPH radical scavenging (%); (B) superoxide anion radical scavenging (%); (C) hydroxyl radical scavenging ability (%); (D) reducing ability (OD 700 nm)).
TABLE 6
| Oxidation resistance index | Whole cell (IC) | Cell extract (CFE) |
| DPPH radical scavenging (%) | 10.36±1.20 | 2.52±1.39 |
| Superoxide anion radical scavengingRemoval rate (%) | 13.47±0.26 | 9.00±1.05 |
| Hydroxyl radical scavenging ability (%) | 26.25±4.99 | 16.17±6.37 |
| Reducing ability (OD 700 nm) | 0.11±0.03 | 0.11±0.01 |
Example 3:
use of lactobacillus johnsonii for the preparation of an antioxidant and immunity enhancing product:
the articles of manufacture of the present application are useful for enhancing the antioxidant capacity and immunity of an animal, thereby reducing the induction of disease in the animal. The lactobacillus johnsonii application data are as follows:
animal experiments and in vivo antioxidant and immunity capacity determination:
1. probiotics liquid preparation
When the screened probiotics are used for preparing the probiotic preparation, the strains are firstly inoculated into the corresponding culture medium to be activated for three generations. Preparing probiotic liquid by using a 20L automatic fermentation tank, calculating the viable count of the produced bacterial liquid by using a plate counting method, wherein the viable count of Lactobacillus johnsonii RS-7 in a sample is 1 multiplied by 10 10 CFU/g.
2. Animal experiment treatment
Selecting 12 healthy weaned piglets with average weight of 6.9-7.1kg and 28 days old. According to the principle that the weight is similar to the proportion of male and female, the test piglets are randomly divided into 2 groups, each group is provided with 1 circle, each circle is provided with 6 heads, and the male and female are respectively half. The control group is fed with basic diet, and the test group adds lactobacillus into the basic diet to adjust the viable count in the word material to be 1 multiplied by 10 7 CFU/g, 3 days for the pre-feeding period, 28 days for the formal trial period. All the pigs testedAccording to the conventional word-raising management of the pig farm, immunization, insect expelling and disinfection procedures during the test are all carried out according to the normal procedures of the pig farm. And after the test feeding is finished, 5mL of anterior vena cava is sampled, the blood is put into a blood tube for coagulation, and the blood is centrifuged for 10min at 3000r/min, and is preserved at the temperature of minus 20 ℃ after serum is separated.
3. Antioxidant capacity assay
The collected piglet serum samples are detected by using an antioxidant capacity index detection kit, and four indexes are measured in total: total antioxidant capacity (T-AOC), total superoxide dismutase (SOD), glutathione peroxidase (GSH-PX), malondialdehyde (MDA).
4. Immunocompetence assay
The determination method of the contents of the serum IgA, igG, igM, IL-1, the IL-10 and the sIgA is determined by an enzyme-linked immunosorbent assay (ELISA), and specific operations are performed according to the instruction of a kit;
detection of antioxidant and immunity of Lactobacillus plantarum RS-7 after application to animals:
the T-SOD, CAT, GSH-Px and T-AOC in serum are detected as indexes of the oxidation resistance of organisms, and MDA is an oxidation damage index. IgA, SIgA, igM and IgG are indicators of the immune level of the body.
Influence of RS-7 on antioxidant capacity of piglets:
the total superoxide dismutase (T-SOD), catalase (CAT), glutathione peroxidase (GSH-PX) and total antioxidant capacity (T-AOC) activities in the serum of the piglets of the experimental group (feeding lactobacillus RS-7 group) are all obviously higher than those of the control group (P <0.05or 0.01). Whereas the serum Malondialdehyde (MDA) content in the experimental group was lower than in the control group. The lactobacillus RS-7 is shown to improve the antioxidant capacity of weaned piglets. The data are shown in Table 7, FIG. 6 ((A) Total superoxide dismutase (T-SOD); (B) Total antioxidant capacity (T-AOC); (C) Catalase (CAT); (D) glutathione peroxidase (GSH-PX); (E) Malondialdehyde (MDA); X indicates very significant differences (P < 0.05); X indicates very significant differences (P < 0.01)).
TABLE 7
Note that: * Indicates that the difference is extremely significant (P < 0.05), and indicates that the difference is extremely significant (P < 0.01).
Effect of RS-7 on immunocompetence of piglets:
the serum of piglets in the experimental group (feeding lactobacillus RS-7 group) contains higher amounts of immunoglobulin A (IgA), secretory immunoglobulin A (IgA), immunoglobulin M (IgM) and immunoglobulin G (IgG) than those in the control group, wherein the experimental group contains significantly higher amounts of immunoglobulin A (IgA), immunoglobulin G (IgG) and secretory immunoglobulin A (IgA) than those in the control group (P <0.05or 0.01). The lactobacillus RS-7 is shown to improve the immunocompetence of weaned pigs. The data are shown in Table 8 and FIG. 7 (note: A) for immunoglobulin A (IgA), B) for immunoglobulin M (IgM), C for immunoglobulin G (IgG), D for secretory immunoglobulin A (IgA), where P <0.05 is very significant and P <0.01 is very significant.
TABLE 8
| Group of | IgA(g/L) | IgM(g/L) | IgG(g/L) | SIg(μg/mL) |
| Control group | 0.91±0.10 | 0.31±0.09 | 2.41±0.46 | 0.24±0.05 |
| Experimental group | 1.04±0.10* | 0.40±0.08 | 4.55±0.60** | 0.36±0.07** |
Note that: * Indicates that the difference is extremely significant (P < 0.05), and indicates that the difference is extremely significant (P < 0.01).
Although embodiments of the present invention have been disclosed above, it is not limited to the details and embodiments shown and described, it is well suited to various fields of use for which the invention would be readily apparent to those skilled in the art, and accordingly, the invention is not limited to the specific details and illustrations shown and described herein, without departing from the general concepts defined in the claims and their equivalents.
Claims (7)
1. Use of lactobacillus johnsonii for the preparation of an antioxidant and immunity enhancing product, wherein the product is a microbial agent or an animal feed.
2. Use of lactobacillus johnsonii according to claim 1 for the preparation of an antioxidant and immunity enhancing product, wherein the viable count of lactobacillus johnsonii in the animal feed is 1 x 10 7 ～2.5×10 7 CFU/g。
3. Use of lactobacillus johnsonii according to claim 1 for the preparation of an antioxidant and immunity enhancing product, wherein the animal feed is a piglet feed.
4. Use of lactobacillus johnsonii according to claim 1 for the preparation of an antioxidant and immunity enhancing product, wherein the bacterial suspension and the cell extract of lactobacillus johnsonii each have a certain DPPH free radical, superoxide anion free radical, hydroxyl radical scavenging capacity and a certain reducing capacity.
5. Use of lactobacillus johnsonii according to claim 1 for the preparation of an antioxidant and immunity enhancing product, wherein the lactobacillus johnsonii is classified under the name Lactobacillus johnsonii RS-7 and has been deposited with the chinese collection of typical cultures under the accession number cctccc No: m20221532.
6. Use of lactobacillus johnsonii according to claim 1 for the preparation of an antioxidant and immunity enhancing product, wherein the lactobacillus johnsonii is derived from fresh pig manure.
7. Use of lactobacillus johnsonii according to claim 1 for the preparation of an antioxidant and immunity enhancing product, wherein the sequence of 16S rDNA of lactobacillus johnsonii is shown in SEQ ID No. 1.
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