CN116004420A - Lactobacillus plantarum NHE-LpB11 and application thereof - Google Patents
Lactobacillus plantarum NHE-LpB11 and application thereof Download PDFInfo
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- CN116004420A CN116004420A CN202210868552.9A CN202210868552A CN116004420A CN 116004420 A CN116004420 A CN 116004420A CN 202210868552 A CN202210868552 A CN 202210868552A CN 116004420 A CN116004420 A CN 116004420A
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- lactobacillus plantarum
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Abstract
The invention provides a lactobacillus plantarum NHE-LpB11 and application thereof, belonging to probiotics for livestock feeding, wherein the probiotics are separated from intestinal tracts of binary sows and are preserved in China general microbiological culture collection center (CGMCC) with the preservation number of 24431 and the preservation date of 2022, 2 and 28. The strain has strong antibacterial performance and strong stability of microecological preparation. Improving the reproductive performance of sows, reducing the production cost, improving the economic benefit and having good application prospect.
Description
Technical Field
The invention relates to probiotics for livestock feeding, in particular to a probiotic lactobacillus plantarum NHE-LpB11 and application thereof.
Background
In recent years, the pig raising industry in China rapidly develops, and more farms develop towards large scale and intensive directions. However, there are still some problems yet to be solved in the pig raising industry, and how to effectively improve the reproductive capacity of sows is one of the problems yet to be solved.
The reproductive capacity of a sow refers to the fertility function of the sow itself and the capacity of the offspring of fertility, which is not only related to the economic income of a farm, but also closely related to whether the pig industry is developing rapidly. There are many factors affecting the reproductive capacity of sows, and among them, feed feeding is a very important one of all factors. In the sow feeding process, if nutrients which are expected to be suitable for the physiological needs cannot be provided for the sow, the reproductive performance of the sow can be seriously affected.
The microecological preparation is used as a novel green feed additive without pollution, residue and any toxic or side effect, has unique advantages which are incomparable with antibiotics in the aspects of regulating the balance of animal intestinal flora, activating animal immune system, promoting animal healthy growth and the like, and therefore, the microecological preparation suitable for improving the reproductive performance of sows has important significance.
Disclosure of Invention
The invention aims to provide lactobacillus plantarum NHE-LpB11 and application thereof in promoting reproductive performance of sows.
In order to achieve the above purpose, the present invention provides the following technical solutions:
the invention provides a lactobacillus plantarum NHE-LpB11, which is classified and named as lactobacillus plantarum (Lactobacillus plantarum) NHE-LpB11, and is preserved in China general microbiological culture collection center (CGMCC) No.24431 in 2/28/2022, and the preservation address is North Chenxi Lu No. 1/3 in the Korean region of Beijing city.
Preferably, the lactobacillus plantarum NHE-LpB is a lactobacillus plantarum strain separated from intestinal contents of healthy binary sows, and is determined to be lactobacillus plantarum through colony morphology observation and molecular biological identification, and the 16S rDNA sequence of the lactobacillus plantarum strain is shown as SEQ ID NO. 1.
Preferably, the lactobacillus plantarum NHE-LpB has the following microbiological characteristics: the strain can grow well on MRS flat plate, and is cultured for 48 hours to form circular colony, the diameter of the colony is 2-3mm, the colony is milky white, the colony is smooth and opaque, the middle is provided with a bulge, the edge is neat, the colony is in a form as shown in figure 1, the micro-form is as shown in figure 2, the colony is gram positive, the colony is short-rod long-chain, has no spore and facultative anaerobic, the proper growth temperature range is 25-45 ℃, the optimal growth temperature is 30-37 ℃, the growth pH is 3.0-8.0, and the optimal pH is 4.5-6.5.
The culture method of the lactobacillus plantarum NHE-LpB11 comprises the following steps:
3mL of lactobacillus plantarum NHE-LpB seed solution (the viable bacteria concentration is 109 CFU/mL) is taken and inoculated into 300mL of shake flask fermentation medium for shake flask fermentation culture; and (3) after the shake flask fermentation is finished, fermenting and culturing in a fermentation tank, and inoculating 300mL of shake flask fermentation seed liquid into a fermentation medium in a 50L fermentation tank for fermenting and culturing, wherein the liquid amount of the 50L fermentation tank is 35L fermentation medium.
The shake flask fermentation medium consists of the following components: 0.5 to 4 percent of sucrose, 0.5 to 2.5 percent of glucose, 0.5 to 3.0 percent of yeast extract powder, 0.5 to 2.5 percent of beef extract powder, 0.01 to 0.5 percent of magnesium chloride, 0.01 to 1.0 percent of calcium carbonate, 0.01 to 0.5 percent of manganese sulfate and the balance of;
preferably, it is: 2% of sucrose, 0.5% of glucose, 1% of yeast extract powder, 1% of beef extract powder, 0.2% of magnesium chloride, 0.1% of calcium carbonate, 0.02% of manganese sulfate and the balance of water.
The shaking flask fermentation conditions are as follows: the inoculation amount is 1% (volume ratio), the fermentation temperature is 35 ℃, the pH is 6.8, the fermentation time is 200r/min, and the fermentation time is 14h.
The culture medium components and the fermentation conditions of the 50L fermentation tank are as follows: 1.0 to 4.0 percent of soft white sugar, 0.2 to 2.0 percent of glucose, 0.5 to 3.5 percent of yeast extract, 0.5 to 2.5 percent of beef extract powder, 0.5 to 2.5 percent of corn dry powder, 0.01 to 0.5 percent of magnesium chloride, 0.01 to 1.0 percent of calcium carbonate, 0.01 to 0.5 percent of manganese sulfate, 0.05 to 0.2 percent of tween-80 and the balance of water.
Preferably, it is: 1.5% of soft white sugar, 0.5% of glucose, 1% of yeast extract, 0.5% of beef extract powder, 0.5% of corn dry powder, 0.2% of magnesium chloride, 0.1% of calcium carbonate, 0.02% of manganese sulfate, 0.1% of tween-80 and the balance of water.
The shaking flask fermentation conditions are as follows: the liquid amount of the 50L fermentation tank is 35L culture medium, the tank pressure is controlled to be 0.05-0.06MPa, the inoculation amount is 300mL, the fermentation temperature is 35 ℃, the fermentation time is 18h, the pH value is 6.0, and the stirring rotating speed is 200r/min.
The invention provides application of lactobacillus plantarum NHE-LpB11 in preparation of broad-spectrum bactericides.
Preferably, the broad-spectrum bactericide has a bactericidal spectrum of erysipelas, pseudomonas aeruginosa, klebsiella pneumoniae, listeria immitis, streptococcus pneumoniae, enteropathogenic escherichia coli, staphylococcus aureus, salmonella typhi, salmonella, clostridium perfringens, proteus pensonii, aeromonas hydrophila and vibrio parahaemolyticus.
The invention provides application of lactobacillus plantarum NHE-LpB11 in preparation of a microecological preparation for promoting reproductive performance of sows.
Preferably, the effective viable count of lactobacillus plantarum NHE-LpB11 in the microecological preparation is more than 1.2X10 11 CFU/g。
Preferably, the sow reproductive performance includes average daily feed intake, litter size, average birth weight of piglets, constipation rate of pregnant sows, constipation rate of lactating sows, auxiliary yield and incidence of birth canal inflammation of sows.
The invention provides a microecological preparation for promoting the reproductive capacity of sows, which is characterized by comprising the following steps:
(1) Centrifuging lactobacillus plantarum NHE-LpB11 fermentation liquor to obtain lactobacillus plantarum NHE-LpB11 active bacterial sludge;
(2) Uniformly mixing the lactobacillus plantarum NHE-LpB11 active bacterial mud and water according to the weight ratio of 1:1 to obtain high-concentration bacterial suspension;
(3) Uniformly mixing the thallus suspension and a protective agent according to the weight ratio of 1:2.2 to prepare microcapsule wet powder;
(4) Wet granules are prepared by a granulator, then the wet granules are uniformly mixed with a coating agent solution, and the mixture is put into a fluidized drying bed for drying and coating, so as to obtain the NHE-LpB11 microecological preparation.
Preferably, the protective agent comprises the following components: 40% of corn starch, 2% of sodium carboxymethyl cellulose, 2% of glucan, 2.4% of inulin, 15% of skimmed milk powder, 10% of gelatinized modified starch, 5% of sucrose, 2% of peptone, 2% of trehalose, 5% of glycerol, 5% of microcrystalline cellulose and the balance of water.
Preferably, the coating agent solution comprises the following components: 10% of sodium alginate, 6.5% of hydroxymethyl cellulose, 8.5% of chitosan, 4% of mannans and the balance of water.
Preferably, the sow reproductive performance includes average daily feed intake, litter size, average birth weight of piglets, constipation rate of pregnant sows, constipation rate of lactating sows, auxiliary yield and incidence of birth canal inflammation of sows.
The beneficial effects of the invention are as follows:
1. the invention provides a lactobacillus plantarum NHE-LpB11 which has higher storage stability and can meet the use requirement; the strain has higher acid resistance, can resist gastric acid and can smoothly reach intestinal tracts to play a role; the survival rate is 97.60 percent after the strain is treated in a solution with 0.3 percent of bile salt solubility for 2 hours, which shows that the strain has higher bile salt resistance and can resist the bile salt in duodenal juice, so that the strain can reach the intestinal tract to play a role.
2. The lactobacillus plantarum NHE-LpB provided by the invention has excellent antibacterial property, can obviously inhibit the growth of erysipelas suis, pseudomonas aeruginosa, klebsiella pneumoniae, listeria immitis, streptococcus pneumoniae, enteropathogenic escherichia coli, staphylococcus aureus, salmonella typhi, salmonella, bacillus perfringens, proteus pensonii, aeromonas hydrophila and vibrio parahaemolyticus, and has broad-spectrum antibacterial property. Therefore, the feed quality stability can be ensured when the feed additive is used in the feed.
3. The microecological preparation prepared from the lactobacillus plantarum NHE-LpB11 can improve the reproductive performance of sows, including average daily feed intake, litter size, healthy litter size, average birth weight of piglets, constipation rate of pregnant sows, constipation rate of lactating sows, auxiliary yield and incidence of birth canal inflammation.
Drawings
FIG. 1 is a colony morphology of Lactobacillus plantarum NHE-LpB11 on MRS medium.
FIG. 2 is a gram stain of Lactobacillus plantarum NHE-LpB strain.
Detailed Description
The examples are presented for better illustration of the invention, but the invention is not limited to the examples. Those skilled in the art will appreciate that various modifications and adaptations of the embodiments described above are possible in light of the above teachings and are intended to be within the scope of the invention.
Unless otherwise indicated, all chemical reagents used in the examples were conventional commercial reagents, and the technical means used in the examples were conventional means well known to those skilled in the art.
The percentage "%" referred to in the present invention refers to mass percent unless otherwise specified; however, the percentage of the solution, unless otherwise specified, refers to the grams of solute contained in 100mL of solution.
Example 1
Isolation, screening and identification of Lactobacillus plantarum NHE-LpB11
1. Separation and purification of lactic acid bacteria
(1) Taking the healthy binary sows to slaughter, taking intestinal tracts, carrying the fresh-keeping to a laboratory, weighing 10g of a sampling product under the aseptic condition, placing the sampling product in a triangular flask containing 90mL of sterilized normal saline, vibrating the sampling product at the constant temperature of 37 ℃ for 1h, sequentially diluting the sampling product to 100 ten thousand times by adopting a 10-fold ratio dilution method, and selecting three dilutions of 1 ten thousand times, 10 ten thousand times and 100 ten thousand times;
(2) Absorbing 0.1mL, coating the solution on an improved MRS agar plate, inversely culturing for 48 hours at 37 ℃ after the plate is coated, picking a suspected lactobacillus colony with obvious calcium dissolving ring and larger than 5mm by using an inoculating loop, carrying out streak separation culture on the MRS agar plate, picking a colony with better separation effect after 48 hours culture, transferring the colony onto an MRS agar inclined plane by using the inoculating loop for pure culture, repeating the pure culture for 3 times, suspending strain cells in a 20% glycerol solution, and storing in a refrigerator at-80 ℃ for later use.
2. Observation of colony morphology
According to the size of the calcium dissolving ring in the step 1, the strains with strong acid production capacity are preliminarily screened out, and the strains can be preliminarily determined to be lactobacillus, and the total number of the strains is 137. Activating the glycerol tube strain stored in the step 1 for 2-3 times by using an MRS agar plate, inoculating the glycerol tube strain into an MRS broth culture medium, performing shake culture at the constant temperature of 37 ℃ for 180r/min for 18-20h, taking a clean glass slide for gram staining, performing microscopic examination, observing the microscopic morphology of the strain, and selecting non-spore bacteria with positive gram staining as standby.
3. Preparation of lactic acid bacteria suspension and fermentation broth
Culturing the lactobacillus obtained in the step 1 on an MRS agar plate at 37 ℃ for 48 hours, picking single colony from the plate, and shake culturing for 24 hours at 37 ℃ and 180r/min in 100mL MRS liquid culture medium to obtain lactobacillus suspension for later use; and continuing to perform shake culture at 37 ℃ and 180r/min for 96 hours to obtain lactobacillus fermentation liquor for later use.
4. Screening of lactic acid bacteria for acid producing ability
Inoculating the lactobacillus suspension obtained in the step 3 into MRS liquid culture medium according to an inoculum size of 1% (volume ratio), culturing for 12 hours at 37 ℃, screening out lactobacillus with the pH of fermentation liquor being less than 3.8 for 12 hours, and obtaining 23 strains of lactobacillus in total.
5. Screening of bacteriostatic lactic acid bacteria
Collecting pathogenic bacteria (such as erysipelas, pseudomonas aeruginosa, klebsiella pneumoniae, listeria monocytogenes, streptococcus pneumoniae, enteropathogenic Escherichia coli, staphylococcus aureus, salmonella typhi, salmonella, bacillus perfringens, proteus pensonii, aeromonas hydrophila, and Vibrio parahaemolyticus) with a concentration of 10 9 2mL of CFU/mL bacterial suspension is added into a pathogenic bacteria culture medium which is filled with 200mL of bacterial suspension and cooled to about 45 ℃ after sterilization, then 10mL of non-solidified bacteria-carrying culture medium is sucked, and the bacteria-carrying culture medium is transferred onto a nutrient agar plate poured with 10mL of bottom plate, so that a plurality of pathogenic bacteria plates are prepared. Each pathogen nutrient agar plate on the ultra-clean workbench is clamped with 1 sterilized oxford cup (a round small tube with the inner diameter of 6mm, the outer diameter of 8mm and the height of 10mm, 200 mu L of liquid can be added into the tube, and two ends of the tube are smooth) and placed on the plate, so that the plate is in contact with a culture medium without gaps, after a few minutes, 200 mu L of suspected lactobacillus fermentation liquor (obtained in the step 3) is respectively sucked into the oxford cup, and the oxford cup is cultivated at the constant temperature of 37 ℃ for 24 hours. Each strain is subjected to at least 3 repetitions, and the size of a bacteriostasis zone is observed and measured, wherein 6 strains with large bacteriostasis zones are respectively marked as B5, B11, B39, B68, B111 and B132.
6. Lactic acid bacteria stability screening
Inoculating the lactobacillus suspension prepared in the step 3 into a sterile MRS broth culture medium according to an inoculum size of 1% (v/v), carrying out shaking culture for 48 hours at 37 ℃ and 180r/min with a liquid loading amount of 300mL/1000mL, then filling the fermentation liquor into a closed bottle, detecting the viable count of the lactobacillus in the fermentation liquor every 2 days, continuously detecting for 30 days, screening out the viable count of the lactobacillus in the B11 fermentation liquor, and taking the viable count of the lactobacillus in the fermentation liquor as the strain for the next study, wherein the survival rate of the lactobacillus in the fermentation liquor for 30 days is 100%.
7. Identification of strain species
The strain B11 is subjected to morphological and molecular identification, the strain B11 can well grow on an MRS flat plate and is cultured for 48 hours to form circular colonies, the diameters of the colonies are 2-3mm, the colonies are milky white, smooth and opaque, the middle is provided with a bulge, the edges are tidy, the colony morphology is shown as figure 1, the microscopic morphology is shown as figure 2, the colony is gram positive, the colony is short-rod long-chain and spore-free, facultative anaerobic, the growth proper temperature is 25-45 ℃, the optimal growth temperature is 30-37 ℃, the growth pH is 3.0-8.0, and the optimal pH is 4.5-6.5. And extracting the genome DNA of the B11 strain by using a kit for extracting the bacterial DNA. Sequencing the 16S rDNA gene fragment of the B11 strain by using the primers F and R, wherein the obtained sequence is shown as SEQ ID NO. 1, and comparing the measured sequence with the 16S rDNA sequence in GenBank by BLAST analysis, so that the homology of the strain B11 and lactobacillus plantarum can be found to reach 99.93 percent. The strain B11 is determined to be lactobacillus plantarum according to the morphological characteristics and 16S rDNA characteristics of the strain B11Lactobacillus plantarum) And is formally numbered as NHE-LpB11.
8. Preservation of strains
The lactobacillus plantarum obtained by separation, purification and screeningLactobacillus plantarum) NHE-LpB11 was preserved in China general microbiological culture Collection center (CGMCC) at 28/2/2022: the institute of microbiology, national institute of sciences, no. 3, north chen west way 1, region of korea, beijing city, postal code: 100101 The preservation number is CGMCC NO.24431, and the classification and naming are carried out on lactobacillus plantarumLactobacillus plantarum）。
Wherein, the composition of the modified MRS agar culture medium is as follows: 1.0% of peptone, 0.5% of sodium acetate, 1.0% of beef extract, 2% of glucose, 0.5% of yeast extract, 0.1% of Tween 80, 0.2% of K2HPO4, 0.058% of MgSO4, 0.2% of diammonium citrate, 0.025% of MnSO4, 1.8% of agar, 1% of calcium carbonate, the balance of water and pH 7.0+/-0.2.
The composition of MRS agar medium is: 1.0% of peptone, 0.5% of sodium acetate, 1.0% of beef extract, 2% of glucose, 0.5% of yeast extract, 0.1% of Tween 80, 0.2% of K2HPO4, 0.058% of MgSO4, 0.2% of diammonium citrate, 40.025% of MnSO, 1.8% of agar and the balance of water, wherein the pH is 7.0+/-0.2.
The composition of the MRS broth medium was: 1.0% of peptone, 0.5% of sodium acetate, 1.0% of beef extract, 2% of glucose, 0.5% of yeast extract, 0.1% of Tween 80, 0.2% of K2HPO4, 0.058% of MgSO4, 0.2% of diammonium citrate, 40.025% of MnSO and the balance of water, and the pH value is 7.0+/-0.2.
The pathogen culture mediums are respectively as follows: the erysipelas pig bacillus, the listeria monocytogenes and the streptococcus pneumoniae are TSA+5% defibrinated sheep blood agar;
pseudomonas aeruginosa, klebsiella pneumoniae, enteropathogenic escherichia coli, staphylococcus aureus, salmonella typhi, salmonella, proteus penmansoni and aeromonas hydrophila are used as nutrient agar culture mediums;
vibrio parahaemolyticus is TCBS culture medium, and Bacillus perfringens is tryptone-sulfite-cycloserine agar culture medium.
The composition of TSA+5% defibrinated sheep blood agar is: 1.5% of tryptone, 0.5% of soyase, 0.5% of sodium chloride, 1.5% of agar, the balance of water, and pH 7.2+/-0.2, and 5% of defibrinated sheep blood is added when the mixture is cooled to 50 ℃ during use.
The nutrient agar medium consists of: 1% of peptone, 0.3% of beef extract, 2% of agar, 0.5% of NaCl and the balance of water, and the pH value is 7.2+/-0.2.
The composition of the TCBS agar medium was: yeast powder 0.5%, peptone 1%, sodium thiosulfate 1%, sodium citrate 1%, ox gall powder 0.5%, sodium taurocholate 0.3%, sucrose 2%, sodium chloride 1%, ferric citrate 0.1%, thymol blue 0.0004%, agar 1.5% and pH 8.6+ -0.1.
The composition of the tryptone-sulfite-cycloserine agar medium was: 1.5% of tryptone, 0.5% of soyase, 0.5% of yeast powder, 0.1% of sodium metabisulfite, 0.1% of ferric ammonium citrate, 2% of agar and the balance of water, wherein pH 7.6+/-0.2 is adopted, and 20mL/250mL of filtered and sterilized 0.5% D-cycloserine solution is added when the solution is cooled to 50 ℃ during use.
Example 2
Preparation of lactobacillus plantarum NHE-LpB11 fermentation broth
(1) Collecting seed liquid of Lactobacillus plantarum NHE-LpB11 (preservation number is CGMCC No. 24431)(viable bacteria concentration is 10) 9 CFU/mL) 3mL, inoculating in 300mL shake flask fermentation medium for shake flask fermentation culture; and (3) after the shake flask fermentation is finished, fermenting and culturing in a fermentation tank, and inoculating 300mL of shake flask fermentation seed liquid into a fermentation medium in a 50L fermentation tank for fermenting and culturing, wherein the liquid amount of the 50L fermentation tank is 35L fermentation medium. After fermentation, detecting the viable count of the fermentation broth to be 4.7X10 10 CFU/mL。
The shake flask fermentation medium consists of the following components:
2% of sucrose, 0.5% of glucose, 1% of yeast extract powder, 1% of beef extract powder, 0.2% of magnesium chloride, 0.1% of calcium carbonate, 0.02% of manganese sulfate and the balance of water.
The shaking flask fermentation conditions are as follows: the inoculation amount is 1% (volume ratio), the fermentation temperature is 35 ℃, the pH is 6.8, the fermentation time is 200r/min, and the fermentation time is 14h.
The culture medium components and the fermentation conditions of the 50L fermentation tank are as follows:
1.5% of soft white sugar, 0.5% of glucose, 1% of yeast extract, 0.5% of beef extract powder, 0.5% of corn dry powder, 0.2% of magnesium chloride, 0.1% of calcium carbonate, 0.02% of manganese sulfate, 0.1% of tween-80 and the balance of water.
The shaking flask fermentation conditions are as follows: the liquid amount of the 50L fermentation tank is 35L culture medium, the tank pressure is controlled to be 0.05-0.06MPa, the inoculation amount is 300mL, the fermentation temperature is 35 ℃, the fermentation time is 18h, the pH value is 6.0, and the stirring rotating speed is 200r/min.
Example 3
Preparation of lactobacillus plantarum NHE-LpB11 microecological preparation
Preparing NHE-LpB fermentation liquor by using lactobacillus plantarum NHE-LpB11 according to the method of example 2, centrifuging the fermentation liquor at 13000r/min for 10min to obtain lactobacillus plantarum NHE-LpB11 active bacterial sludge, and uniformly mixing the lactobacillus plantarum active bacterial sludge and water according to the weight ratio of 1:1 to obtain a high-concentration bacterial suspension. Uniformly mixing lactobacillus plantarum NHE-LpB11 thallus suspension and a protective agent according to the weight ratio of 1:2.2, preparing into microcapsule wet powder, preparing into wet particles by a granulator, uniformly mixing with a coating agent solution, putting into a fluidized drying bed, and drying and coating to obtain the preparation of the NHE-LpB11 microecological preparation, wherein the effective viable count reaches 1.2 multiplied by 10 11 CFU/g。
The components of the protective agent are as follows: 40% of corn starch, 2% of sodium carboxymethyl cellulose, 2% of glucan, 2.4% of inulin, 15% of skimmed milk powder, 10% of gelatinized modified starch, 5% of sucrose, 2% of peptone, 2% of trehalose, 5% of glycerol, 5% of microcrystalline cellulose and the balance of water.
The components of the coating agent solution are as follows: 10% of sodium alginate, 6.5% of hydroxymethyl cellulose, 8.5% of chitosan, 4% of mannans and the balance of water.
Example 4
Probiotic verification of lactobacillus plantarum NHE-LpB11
(1) Collecting pathogenic bacteria (such as erysipelas, pseudomonas aeruginosa, klebsiella pneumoniae, listeria monocytogenes, streptococcus pneumoniae, enteropathogenic Escherichia coli, staphylococcus aureus, salmonella typhi, salmonella, bacillus perfringens, proteus pensonii, aeromonas hydrophila, and Vibrio parahaemolyticus) with a concentration of 10 9 2mL of CFU/mL bacterial suspension is added into a pathogenic bacteria culture medium which is filled with 200mL of bacterial suspension and cooled to about 45 ℃ after sterilization, then 10mL of non-solidified bacteria-carrying culture medium is sucked, and the bacteria-carrying culture medium is transferred onto a nutrient agar plate poured with 10mL of bottom plate, so that a plurality of pathogenic bacteria plates are prepared.
(2) 1 sterilized oxford cup (round small tube with inner diameter of 6mm, outer diameter of 8mm and height of 10 mm) is clamped by sterile forceps on each pathogen nutrient agar plate on an ultra-clean workbench, 200 mu L of liquid can be added into the tube, two ends of the tube are smooth) is placed on the plate, the plate is contacted with a culture medium without gaps, 200 mu L of the fermentation liquor prepared in the preserved example 2 is respectively dripped into each small tube after 10 minutes, the fermentation liquor is not overflowed, the culture is carried out for 36-96 hours at 37 ℃, and then the diameter of a bacteriostasis zone is measured. Three replicates per experiment were averaged and the results are shown in table 1.
The pathogen culture mediums are respectively as follows: the erysipelas pig bacillus, the listeria monocytogenes and the streptococcus pneumoniae are TSA+5% defibrinated sheep blood agar;
pseudomonas aeruginosa, klebsiella pneumoniae, enteropathogenic escherichia coli, staphylococcus aureus, salmonella typhi, salmonella, proteus penmansoni and aeromonas hydrophila are used as nutrient agar culture mediums;
vibrio parahaemolyticus is TCBS culture medium, and Bacillus perfringens is tryptone-sulfite-cycloserine agar culture medium.
TSA+5% defibrinated sheep blood agar composition was: 1.5% of tryptone, 0.5% of soyase, 0.5% of sodium chloride, 1.5% of agar, the balance of water, and pH 7.2+/-0.2, and 5% of defibrinated sheep blood is added when the mixture is cooled to 50 ℃ during use.
The nutrient agar medium consists of: 1% of peptone, 0.3% of beef extract, 2% of agar, 0.5% of NaCl and the balance of water, and the pH value is 7.2+/-0.2.
The TCBS agar medium consists of: yeast powder 0.5%, peptone 1%, sodium thiosulfate 1%, sodium citrate 1%, ox gall powder 0.5%, sodium taurocholate 0.3%, sucrose 2%, sodium chloride 1%, ferric citrate 0.1%, thymol blue 0.0004%, agar 1.5% and pH 8.6+ -0.1.
The composition of the tryptone-sulfite-cycloserine agar medium is as follows: 1.5% of tryptone, 0.5% of soyase, 0.5% of yeast powder, 0.1% of sodium metabisulfite, 0.1% of ferric ammonium citrate, 2% of agar and the balance of water, wherein pH 7.6+/-0.2 is adopted, and 20mL/250mL of filtered and sterilized 0.5% D-cycloserine solution is added when the solution is cooled to 50 ℃ during use.
TABLE 1 antibacterial Effect of Lactobacillus plantarum NHE-LpB11 on pathogenic bacteria
| Pathogenic bacteria | Antibacterial diameter (mm) |
| Erysipelothrix rhusiopathiaeErysipelas suis | 24.21 |
| Pseudomonas aeruginosaPseudomonas aeruginosa | 22.31 |
| Klebsiella pneumoniaeKlebsiella pneumoniae | 25.13 |
| Listeria monocytogenesListeria ivanovii | 25.32 |
| Streptococcus pneumoniaeStreptococcus pneumoniae | 22.14 |
| Enteric pathogenic escherichia coliEscherichia coli | 27.42 |
| Staphylococcus aureusStBphylococcus Bureus | 28.78 |
| Salmonella typhiSalmonella typhimurium | 29.13 |
| Salmonella bacteriaSalmonella | 31.07 |
| Shigella genusShigella Castellani | 28.32 |
| Clostridium perfringensClostridium perfringen | 33.24 |
| Proteus pensoniiProteus penneri | 18.91 |
| Aeromonas hydrophilaAeromonas hydrophila | 26.32 |
| Vibrio parahaemolyticusVibrio parahaemolyticus | 17.23 |
It can be seen that the lactobacillus plantarum NHE-LpB has remarkable inhibition effects on erysipelas, pseudomonas aeruginosa, klebsiella pneumoniae, listeria immaturus, streptococcus pneumoniae, enteropathogenic escherichia coli, staphylococcus aureus, salmonella typhi, salmonella, clostridium perfringens, proteus peng, aeromonas hydrophila and vibrio parahaemolyticus.
Example 5
Stress resistance verification of lactobacillus plantarum NHE-LpB11 microecological preparation
1. Storage stability verification
Placing the lactobacillus plantarum NHE-LpB11 microecological preparation into a sealed bag, and then placing the sealed bag filled with the lactobacillus plantarum microecological preparation into a constant temperature and humidity incubator, wherein the temperature is set to 37 ℃ and the relative humidity is 60%. The sample is sampled and tested for the effective viable count of the lactobacillus plantarum weekly, and the result shows that the lactobacillus plantarum has an accelerated survival rate of 100 percent and 91 percent after being stored for 21 days and 28 days at 37 ℃ by comparing with the initial effective viable count. The microbial ecological agent prepared by the strain NHE-LpB11 has higher storage stability and can meet the use requirement.
2. Determination of artificial gastric juice resistance:
accurately weighing 10.000g of lactobacillus plantarum NHE-LpB11 microecological preparation sample, placing the sample in 90mL (250 mL triangular flask) of prepared artificial gastric juice, and vibrating at constant temperature of 37 ℃ and 200r/min for 60min; and after the shaking is finished, 10mL of sample solution is taken to adjust the pH value to 7.0, 90mL of physiological saline is added, the shaking is performed for 30min at a constant temperature of 200r/min at 37 ℃, and then the colony culture count of the dilution plate is performed. The results are shown in Table 2. As can be seen from Table 2, the survival rate of the lactobacillus plantarum NHE-LpB11 microecological preparation treated in artificial gastric juice (containing enzyme) with pH of 1.5, pH of 2.0 and pH of 2.5 respectively for 3 hours is over 94 percent, which indicates that the lactobacillus plantarum NHE-LpB11 microecological preparation has higher acid resistance, can resist gastric acid and can smoothly reach intestinal tracts to play a role.
The preparation method of the artificial gastric juice comprises the following steps: preparation of artificial gastric juice referring to preparation method in the "pharmacopoeia of the people's republic of China" 2010 edition, 16.4mL of diluted hydrochloric acid is taken, about 800mL of water and 10g of pepsin are added, shaking is carried out, water is added to be weighed and released into 1000mL, pH values are respectively adjusted to 1.5, 2.0 and 2.5, and microporous filter membrane is sterilized (0.22 μm) for standby.
TABLE 2 survival of Lactobacillus plantarum NHE-LpB11 microecologics 3h after treatment in artificial gastric juice
| Treatment of | pH1.5 | pH2.0 | pH2.5 |
| Initial Activity CFU/mL | 1.25×10 11 | 1.25×10 11 | 1.25×10 11 |
| Post-treatment Activity CFU/mL | 1.18×10 11 | 1.21×10 11 | 1.23×10 11 |
| Survival after treatment% | 94.40 | 96.80 | 98.40 |
3. Determination of bile salt resistance
Accurately weighing 1.000g of lactobacillus plantarum NHE-LpB11 microecological preparation sample, placing the sample in 99mL (250 mL triangular flask) solutions with different bile salt concentrations, wherein the bile salt concentrations are 0.03%, 0.15% and 0.3% respectively, then vibrating at a constant temperature of 37 ℃ and 200r/min for 120min, taking 1mL of sample solution after the vibration is completed, adding 99mL of physiological saline, vibrating at a constant temperature of 37 ℃ and 200r/min for 30min, and then performing colony culture counting on a dilution plate. The results are shown in Table 3. The lactobacillus plantarum NHE-LpB11 microecological preparation is treated in a solution with 0.3% of bile salt solubility for 2 hours, and the survival rate is 97.60%, which shows that the strain has higher bile salt resistance and can resist bile salt in duodenal juice, so that the strain can reach intestinal tracts to play a role.
The preparation method of the bile salt solution with different concentrations comprises the following steps: 1.8mL, 9mL and 18mL of 5% bile salt solution are respectively added into PBS solution with pH of 7.4, the volume is fixed to 300mL, and the PBS solution containing 0.03%, 0.15% and 0.30% of bile salt is uniformly mixed.
The preparation method of the 5% bile salt solution comprises the following steps: accurately weighing 5.0g of bile salt, dissolving with 100mLPBS solution to constant volume, and sterilizing at 121deg.C for 20min.
The preparation method of the PBS solution comprises the following steps: sodium chloride 0.8%, potassium chloride 0.02%, disodium hydrogen phosphate 0.363%, potassium dihydrogen phosphate 0.024%, the balance being water. Adjusting pH to 7.4 with 6mol/L HCl, sterilizing at 121deg.C for 20min, and keeping.
Table 3 survival of NHE-LpB11 micro-ecological formulations after 2h treatment in bile salt solutions of different concentrations
| Treatment of | 0.03% | 0.15% | 0.3% |
| Initial Activity CFU/mL | 1.25×10 11 | 1.25×10 11 | 1.25×10 11 |
| Post-treatment Activity CFU/mL | 1.25×10 11 | 1.25×10 11 | 1.22×10 10 |
| Survival after treatment% | 100 | 100 | 97.60 |
Example 6
Acute toxicity test of Lactobacillus plantarum NHE-LpB11
The safety evaluation of lactobacillus plantarum is carried out by adopting an acute toxicity test and referring to the national standard GB15193.3-2003 maximum tolerance dosage method. 60 common Kunming mice are taken, the male and female mice are half, 18-20g are kept for 1 week conventionally, the mice are subjected to stomach irrigation three times a day, 0.25g/mL lactobacillus plantarum NHE-LpB bacterial liquid (equivalent to 15000mg/kg body weight) is continuously irrigated for 2 weeks, and whether the mice are poisoned or die is observed.
During the test, the mice have good mental state and no poisoning and death phenomena, so that the maximum tolerance dose MTD of the strain of the invention in the acute toxicity test is more than 15000mg/kg, and the strain can be determined to be of a non-toxic grade according to the grading standard, and has higher safety.
Example 7
Effect of Lactobacillus plantarum NHE-LpB11 microecologics on sow reproductive performance
60 long-by-large binary hybrid lactating sows with close gestation and good health condition are selected, and are equally divided into two groups of 30 groups of 6 columns of 5 groups of sows, the test is finished from 15 days before pregnancy to 28 days after weaning of the lactating piglets, the control group is taken as basic ration, and 200 g/ton of lactobacillus plantarum NHE-LpB11 microecologics prepared in the example 3 are added into the test group taken as basic ration. The test pigs adopt a closed colony house, are fed 3 times a day, eat and drink water freely, clean the colony house every day, and feed for 80 days. The feeding management and the immunization program are the same as the daily management of a pig farm, and the pigs are regularly disinfected and found to be ill and treated in time. The test results show that the lactobacillus plantarum NHE-LpB11 microecological preparation remarkably improves the litter size (P < 0.05), the average daily feed intake of the sows in the test group is remarkably increased by 12.37 percent (P < 0.05) compared with that of the sows in the control group, and the average birth weight of piglets is increased by 120g (P < 0.05); the constipation rate of sows in gestation period and sows in lactation period is obviously reduced (P < 0.05), the health level and the autoimmune power of the sows are obviously improved, the auxiliary yield is reduced (P < 0.05), and the occurrence rate of postpartum birth canal inflammation of the sows is effectively reduced (P < 0.05).
TABLE 4 influence of Lactobacillus plantarum NHE-LpB11 microecologics on sow reproductive performance
| Project | Control group | Test group |
| Average daily feed intake of sow (kg/head/day) | 4.93±0.29 b | 5.54±0.39 a |
| Farrowing number (head/nest) | 10.26±1.16 b | 13.37±1.02 a |
| Health number (head/nest) | 8.89±0.23 b | 12.35±0.78 a |
| Average piglet birth weight (kg/head) | 1.31±0.09 b | 1.43±0.05 a |
| Pregnant sow constipation rate (%) | 23.33±0.07 a | 3.34±0.07 b |
| Constipation rate of lactating sow (%) | 36.67±0.32 a | 10.12±0.21 b |
| Yield aid (%) | 33.33±0.45 a | 12.5±2.12 b |
| Incidence of birth canal inflammation (%) | 29.17±0.34 a | 12.5±0.09 b |
Note that: the differences are represented by different letters of the same column, with lower case letters representing the differences being significant (P < 0.05).
Finally, it is noted that the above embodiments are only for illustrating the technical solution of the present invention and not for limiting the same, and although the present invention has been described in detail with reference to the preferred embodiments, it should be understood by those skilled in the art that modifications and equivalents may be made thereto without departing from the spirit and scope of the technical solution of the present invention, which is intended to be covered by the scope of the claims of the present invention.
Claims (10)
1. Lactobacillus plantarum NHE-LpB11, classified and named Lactobacillus plantarum (Lactobacillus plantarum) NHE-LpB11, is preserved in China general microbiological culture Collection center of China Committee for culture Collection of microorganisms for 2 months and 28 days in 2022, and has a preservation number of CGMCC No.24431 and a preservation address of China institute of microbiological culture Collection, national institute No. 3, national institute of sciences 1, north West road, korean, beijing.
2. Use of lactobacillus plantarum NHE-LpB11 as claimed in claim 1 for the preparation of a broad spectrum fungicide.
3. The use according to claim 2, wherein the broad-spectrum fungicide has a bactericidal spectrum of erysipelas, pseudomonas aeruginosa, klebsiella pneumoniae, listeria immitis, streptococcus pneumoniae, enteropathogenic escherichia coli, staphylococcus aureus, salmonella typhi, salmonella, bacillus perfringens, proteus pensonii, aeromonas hydrophila, vibrio parahaemolyticus.
4. Use of lactobacillus plantarum NHE-LpB11 as claimed in claim 1 for the preparation of a probiotic for promoting reproductive performance in sows.
5. The use according to claim 4, wherein the effective viable count of lactobacillus plantarum NHE-LpB11 in the probiotic is greater than 1.2x10 11 CFU/g。
6. The use according to claim 4 wherein the reproductive performance of sows comprises average daily feed intake, litter size, average birth weight of piglets, constipation rate of pregnant sows, constipation rate of lactating sows, aid-rate and incidence of birth canal inflammation.
7. The microecological preparation for promoting the reproductive capacity of the sow is characterized by comprising the following steps of:
(1) Centrifuging lactobacillus plantarum NHE-LpB11 fermentation liquor to obtain lactobacillus plantarum NHE-LpB11 active bacterial sludge;
(2) Uniformly mixing the lactobacillus plantarum NHE-LpB11 active bacterial mud and water according to the weight ratio of 1:1 to obtain high-concentration bacterial suspension;
(3) Uniformly mixing the thallus suspension and a protective agent according to the weight ratio of 1:2.2 to prepare microcapsule wet powder;
(4) Wet granules are prepared by a granulator, then the wet granules are uniformly mixed with a coating agent solution, and the mixture is put into a fluidized drying bed for drying and coating, so as to obtain the NHE-LpB11 microecological preparation.
8. The probiotic formulation of claim 7, wherein the protectant comprises the following components: 40% of corn starch, 2% of sodium carboxymethyl cellulose, 2% of glucan, 2.4% of inulin, 15% of skimmed milk powder, 10% of gelatinized modified starch, 5% of sucrose, 2% of peptone, 2% of trehalose, 5% of glycerol, 5% of microcrystalline cellulose and the balance of water.
9. The probiotic formulation of claim 7, wherein the coating solution comprises the following components: 10% of sodium alginate, 6.5% of hydroxymethyl cellulose, 8.5% of chitosan, 4% of mannans and the balance of water.
10. The microecological formulation of claim 7, wherein the reproductive performance of the sow comprises average daily feed intake, litter size, average birth weight of the piglet, constipation rate of pregnant sows, constipation rate of lactating sows, labor rate and incidence of birth canal inflammation of the sow.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116731939A (en) * | 2023-08-15 | 2023-09-12 | 山东利邦牧业股份有限公司 | Lactobacillus plantarum and application thereof |
| CN117448213A (en) * | 2023-10-24 | 2024-01-26 | 山东宝来利来生物工程股份有限公司 | Lactobacillus plantarum for inhibiting clostridium perfringens and its progeny and application |
| CN117660250A (en) * | 2023-12-08 | 2024-03-08 | 华中农业大学 | Low-temperature lactobacillus and application thereof |
Citations (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101974449A (en) * | 2010-09-13 | 2011-02-16 | 郑州大学 | Lactobacillus plantarum and application thereof |
| CN104789511A (en) * | 2015-05-08 | 2015-07-22 | 内蒙古农业大学 | Lactobacillus plantarum AB-2 with broad-spectrum antifungal properties and application thereof |
| CN104830731A (en) * | 2015-05-08 | 2015-08-12 | 内蒙古农业大学 | Lactobacillus plantarum AB-1 with broad-spectrum bacteriostasis and application thereof |
| CN105132322A (en) * | 2015-09-02 | 2015-12-09 | 广州格拉姆生物科技有限公司 | Lactobacillus plantarum and application thereof |
| KR20160027796A (en) * | 2014-09-02 | 2016-03-10 | 대한민국(농촌진흥청장) | Lactobacillus plantarum jsa22 strain and composition comprising the same |
| CN106399196A (en) * | 2016-11-04 | 2017-02-15 | 北京好实沃生物技术有限公司 | Lactobacillus plantarum HEW-A490 and application thereof |
| CN107312726A (en) * | 2017-05-27 | 2017-11-03 | 武汉科缘生物发展有限责任公司 | One lactobacillus plantarum ZN 3 and application |
| CN107788243A (en) * | 2017-10-31 | 2018-03-13 | 北京小小牧人生物科技有限公司 | A kind of high microsteping sow feed and preparation method and application |
| CN113388551A (en) * | 2021-07-16 | 2021-09-14 | 新希望六和股份有限公司 | Pediococcus pentosaceus NHB-PpA9601 and application thereof |
| CN114437991A (en) * | 2022-03-02 | 2022-05-06 | 青海省畜牧兽医科学院 | Lactobacillus plantarum 160 and application thereof |
-
2022
- 2022-07-21 CN CN202210868552.9A patent/CN116004420B/en active Active
Patent Citations (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101974449A (en) * | 2010-09-13 | 2011-02-16 | 郑州大学 | Lactobacillus plantarum and application thereof |
| KR20160027796A (en) * | 2014-09-02 | 2016-03-10 | 대한민국(농촌진흥청장) | Lactobacillus plantarum jsa22 strain and composition comprising the same |
| CN104789511A (en) * | 2015-05-08 | 2015-07-22 | 内蒙古农业大学 | Lactobacillus plantarum AB-2 with broad-spectrum antifungal properties and application thereof |
| CN104830731A (en) * | 2015-05-08 | 2015-08-12 | 内蒙古农业大学 | Lactobacillus plantarum AB-1 with broad-spectrum bacteriostasis and application thereof |
| CN105132322A (en) * | 2015-09-02 | 2015-12-09 | 广州格拉姆生物科技有限公司 | Lactobacillus plantarum and application thereof |
| CN106399196A (en) * | 2016-11-04 | 2017-02-15 | 北京好实沃生物技术有限公司 | Lactobacillus plantarum HEW-A490 and application thereof |
| CN107312726A (en) * | 2017-05-27 | 2017-11-03 | 武汉科缘生物发展有限责任公司 | One lactobacillus plantarum ZN 3 and application |
| CN107788243A (en) * | 2017-10-31 | 2018-03-13 | 北京小小牧人生物科技有限公司 | A kind of high microsteping sow feed and preparation method and application |
| CN113388551A (en) * | 2021-07-16 | 2021-09-14 | 新希望六和股份有限公司 | Pediococcus pentosaceus NHB-PpA9601 and application thereof |
| CN114437991A (en) * | 2022-03-02 | 2022-05-06 | 青海省畜牧兽医科学院 | Lactobacillus plantarum 160 and application thereof |
Non-Patent Citations (1)
| Title |
|---|
| 庞清刚等: "植物乳杆菌对母猪繁殖性能、仔猪生长性能及免疫水平的影响", 黑龙江畜牧兽医, no. 9, 10 May 2018 (2018-05-10) * |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116731939A (en) * | 2023-08-15 | 2023-09-12 | 山东利邦牧业股份有限公司 | Lactobacillus plantarum and application thereof |
| CN116731939B (en) * | 2023-08-15 | 2023-11-07 | 山东利邦牧业股份有限公司 | Lactobacillus plantarum and application thereof |
| CN117448213A (en) * | 2023-10-24 | 2024-01-26 | 山东宝来利来生物工程股份有限公司 | Lactobacillus plantarum for inhibiting clostridium perfringens and its progeny and application |
| CN117448213B (en) * | 2023-10-24 | 2024-06-04 | 山东宝来利来生物工程股份有限公司 | Lactobacillus plantarum for inhibiting clostridium perfringens and its progeny and application |
| CN117660250A (en) * | 2023-12-08 | 2024-03-08 | 华中农业大学 | Low-temperature lactobacillus and application thereof |
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