CN115053935A - Application of lactobacillus salivarius in breeding of litopenaeus vannamei larvae - Google Patents
Application of lactobacillus salivarius in breeding of litopenaeus vannamei larvae Download PDFInfo
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- CN115053935A CN115053935A CN202210515384.5A CN202210515384A CN115053935A CN 115053935 A CN115053935 A CN 115053935A CN 202210515384 A CN202210515384 A CN 202210515384A CN 115053935 A CN115053935 A CN 115053935A
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- lactobacillus salivarius
- litopenaeus vannamei
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Abstract
The invention discloses an application of lactobacillus salivarius in the cultivation of young litopenaeus vannamei, belonging to the technical field of aquaculture. In order to ensure good probiotic effect of the lactobacillus salivarius on the young litopenaeus vannamei, in the process of breeding the young litopenaeus vannamei, the lactobacillus salivarius fermentation liquor is mixed with the feed and is fed into the young litopenaeus vannamei after being placed for 10min, which is beneficial to promoting the digestion and absorption of protein and increasing the synthesis of antioxidant metabolites and ecdysis endogenous components, thereby obviously improving the growth rate and weight growth rate of the young litopenaeus vannamei, promoting the growth of the young litopenaeus vannamei and better meeting the requirements of the breeding production practice of the young litopenaeus vannamei.
Description
Technical Field
The invention belongs to the technical field of aquaculture, and particularly relates to application of lactobacillus salivarius in improving the growth performance of young litopenaeus vannamei during the process of breeding the young litopenaeus vannamei.
Background
The Litopenaeus vannamei (Litopenaeus vannamei) belongs to shellfish aquatic animals and has a wide distribution range. Salt tolerance, suitable growth salinity of 0.5-40 per mill, suitable growth temperature of 23-30 ℃, suitable growth pH of 7.5-8.5, and better growth with water dissolved oxygen of more than 4.0 mg/L. The culture period from hatching to shrimp growth is generally 100-120 days.
The litopenaeus vannamei is one of the important marine products in China. In 2019, the annual output of Litopenaeus vannamei in China reaches 111.8 ten thousand tons, which accounts for 77.1 percent of the total output of the Litopenaeus vannamei cultured in seawater. However, with the popularization of the high-density intensive culture mode, the shrimp larvae are easily infected by pathogens such as viruses and vibrios in the culture process, so that diseases are frequent. Common diseases in shrimp fry cultivation are white spot syndrome (caused by white spot syndrome virus), acute hepatopancreatic necrosis (caused by vibrio parahaemolyticus), slow growth (shrimp liver enterocytozoon) and the like. Wherein the bacterial diseases mainly comprise Vibrio, in addition to Vibrio parahaemolyticus, Vibrio harveyi, Vibrio anguillarum, Vibrio alginolyticus, Vibrio campylobacter, Vibrio vulnificus, Vibrio cholerae, etc.
At present, the disease prevention and control means applied to shrimp larva cultivation mainly comprises chemical drugs such as antibiotics, and the like, but drug residues and resistance gene transfer generated by the disease prevention and control means further cause food safety problems and environmental pollution problems, and are not beneficial to the health of people and animals. Therefore, it is necessary to develop a method for effectively controlling diseases with environmental protection and safety.
The probiotic preparation is prepared by processing live microorganisms which are beneficial to animals and are harmless through a specific process, and is used for regulating and controlling the micro-ecological balance of a culture water area and aquatic animals. The method does not generate drug residue, does not cause drug-resistant gene transfer, is suitable for large-area use, and does not cause food safety problems, so the method is an effective method which is green, safe and environment-friendly and can replace chemical drugs. Among them, Lactobacillus salivarius is receiving increasing attention as a relatively novel probiotic preparation by virtue of its good probiotic effect.
Lactobacillus salivarius belongs to lactic acid bacteria, and the culture medium is fermentable carbohydrate. No flagellum, no spore production, facultative anaerobic, gram-positive bacteria. It has been reported that lactobacillus salivarius can prevent bacterial diseases of aquatic animals, enhance immunity of aquatic animals, promote growth and survival of aquatic animals, and the like. At present, research and application of lactobacillus salivarius in aquaculture are mainly focused on abalones and/or fishes such as zander, turbot and the like, and domestic application research on improving the growth performance of litopenaeus vannamei fries by lactobacillus salivarius is not reported.
Disclosure of Invention
In order to overcome the defects of the prior art, the invention aims to provide the application of the lactobacillus salivarius in the cultivation of the young litopenaeus vannamei.
The laboratory successfully separates Lactobacillus salivarius GZPH2 from the pickled vegetables sold in Guangzhou, has broad-spectrum antibacterial activity, can inhibit the growth of 39 pathogenic bacteria such as staphylococcus aureus, has no hemolysis phenomenon, does not produce histamine, resists acid and bile salt (CN 104611251A, a lactic acid bacteria strain with broad-spectrum antibacterial activity and application thereof).
The purpose of the invention is realized by the following technical scheme:
the invention provides an application of lactobacillus salivarius in the cultivation of young litopenaeus vannamei.
Furthermore, the lactobacillus salivarius is applied to improving the growth performance of the litopenaeus vannamei.
Furthermore, the lactobacillus salivarius is applied to improving the growth performance of the young litopenaeus vannamei.
Further, the lactobacillus salivarius is applied to promoting the ecdysis and/or the synthesis of secondary metabolites related to the growth of the young litopenaeus vannamei.
Preferably, the lactobacillus salivarius is used for up-regulating the metabolite short peptide and amino acid derivative of the litopenaeus vannamei fry, sesquiterpene, steroid and derivatives thereof, benzene compounds, flavonoid and/or pantothenic acid.
Further, the short peptide is a dipeptide; further at least one of Methionyl-Proline, Valyl-Proline, Tyrosyl-Proline and L-phenylallyl-L-Proline;
further, the amino acid derivative is Glutamate, gamma-methyl ester;
further, the sesquiterpene is farnesyl cysteine;
further, the steroid and the derivative thereof are at least one of 3a,21-Dihydroxy-5b-pregnane-11,20-dione, 3 b-allotetrahydrocortisone, Convalloside and 2-deoxyastasterone;
further, the benzene compound is at least one of N- [ (4-Hydroxy-3-methoxyphenyl) methyl ] octatamide, 6-Gingerol and Adipostatin A;
further, the flavonoid is Formononetin.
The application is realized by mixing lactobacillus salivarius fermentation liquor and feeding the mixed liquor into a litopenaeus vannamei fry culture water body.
Wherein the concentration of Lactobacillus salivarius in water is 10 4 ～10 6 CFU/mL; preferably 10 5 CFU/mL。
The Lactobacillus salivarius is Lactobacillus salivarius (Lactobacillus salivarius) GZPH2, and the preservation unit is as follows: china Center for Type Culture Collection (CCTCC), preservation date: 11/27/2014, storage address: wuhan university in Wuhan City, China, the preservation number: CCTCC NO: m2014598. The strain is disclosed in a patent CN 104611251A, a lactic acid bacterium with broad-spectrum bacteriostatic activity and application thereof.
Further, the concentration of the lactobacillus salivarius fermentation liquor is 8 multiplied by 10 7 ～1×10 8 CFU/mL。
The preparation method of the lactobacillus salivarius fermentation liquor comprises the following steps:
inoculating single lactobacillus salivarius colony in MRS liquid culture medium, and culturing to obtain seed liquid; then inoculating the seed solution into an MRS liquid culture medium in an inoculation amount of 1-5% v/v (preferably 2% v/v), and culturing again to obtain lactobacillus salivarius fermentation liquor; the concentration is 8X 10 7 ～1×10 8 CFU/mL。
The culture condition is that the culture is carried out for 12-36 h under the conditions of 16-40 ℃ and 200rev/min (rpm); further culturing at 37 deg.C and 200rev/min (rpm) for 24 h;
the secondary culture condition is that the culture is carried out for 12-36 h under the condition of standing at 16-40 ℃; further, the cells were cultured at 37 ℃ for 24 hours while standing still.
The lactobacillus salivarius GZPH2 is used for promoting the exuviation of the young shrimps and the synthesis of secondary metabolites related to the growth of the young shrimps, so that the functions of the organism are enhanced, the growth of the young litopenaeus vannamei is promoted, and the method is realized by mixing lactobacillus salivarius fermentation liquor with materials and then feeding the mixture into a young litopenaeus vannamei cultivation water body.
Compared with the prior art, the invention has the following advantages and effects:
in order to ensure good probiotic effect of the lactobacillus salivarius on the young litopenaeus vannamei, in the process of breeding the young litopenaeus vannamei, the lactobacillus salivarius fermentation liquor is mixed with the feed and is fed into the young litopenaeus vannamei after being placed for 10min, which is beneficial to promoting the digestion and absorption of protein and increasing the synthesis of antioxidant metabolites and ecdysis endogenous components, thereby obviously improving the growth rate and weight growth rate of the young litopenaeus vannamei, promoting the growth of the young litopenaeus vannamei and better meeting the requirements of the breeding production practice of the young litopenaeus vannamei.
Drawings
FIG. 1 is a hierarchical clustering heatmap of metabolites with significant differences between the test and blank control groups in example 2; wherein, C7_1, C7_2 and C7_3 in the abscissa represent three replicate samples of the blank control group; l7_1, L7_2, L7_3 represent three replicate samples of the test group. The ordinate represents a significantly different metabolite, showing color in terms of relative expression of the metabolite, the deeper the red the higher the metabolite expression, and the deeper the blue the lower.
FIG. 2 is a KEGG pathway enrichment plot between the blank control group and the test group in example 2; wherein the abscissa represents the pathway name and the ordinate represents the enrichment ratio. The pillar color gradient indicates the significance of the enrichment, the darker the color, the more significant enrichment of the pathway, with the p-value <0.05 labeled as x.
Detailed Description
The present invention will be described in further detail with reference to examples and drawings, but the present invention is not limited thereto.
The test methods in the following examples, in which specific experimental conditions are not specified, are generally performed according to conventional experimental conditions or according to the experimental conditions recommended by the manufacturer. The materials, reagents and the like used are all commercially available reagents and materials unless otherwise specified.
The invention provides application of lactobacillus salivarius fermentation liquor in promoting growth of litopenaeus vannamei larvae.
The present invention will be described in detail below by way of examples.
The media referred to in the examples:
MRS liquid medium (g/L): peptone 20; 20 parts of glucose; soaking beef in powder 8; 5, yeast extract; sodium acetate 5; dipotassium hydrogen phosphate 2; diammonium hydrogen citrate 2; 0.25 of manganese sulfate heptahydrate; magnesium sulfate heptahydrate 0.1; tween-80: 1; MSG (sodium glutamate): 3; sea crystals 3; final pH6.2 + -0.2; subpackaging, sterilizing at 121 deg.C and 0.1MPa for 20min, and storing.
MRS solid medium: adding 5g/L calcium carbonate into MRS liquid culture medium, wherein the gel strength is 1300g/cm and the mass ratio is 1.5% 2 The agar powder is sterilized at 121 deg.C and 0.1MPa for 20min, cooled to 60 deg.C, poured onto sterile plate, solidified, turned over, and stored for use.
The Lactobacillus salivarius used in the examples was Lactobacillus salivarius (Lactobacillus salivarius) GZPH 2.
Example 1 test of Lactobacillus salivarius fermentation broth on Litopenaeus vannamei larvae (PL7-8)
(1) Preparing a litopenaeus vannamei fry (PL 7-8): PL7-8(postlarvae, PL7-8) refers to a 7-8 day old post-litopenaeus vannamei shrimp, and the shrimp fries in the test are all provided by a shrimp fry hatchery in Guangdong province, are consistent in size, are active and healthy, and have no signs of diseases.
(2) Preparation of lactobacillus salivarius fermentation liquor: single colonies were picked from MRS plates using a sterile inoculating loop, inoculated in 50mL MRS liquid medium, and cultured at 37 ℃ under 200rev/min (rpm) for 24h, and used as seed liquid. Then, the seed solution was inoculated at an inoculum size of 2% v/v into a conical flask containing 500mL of MRS liquid medium, and the mixture was left to stand at 37 ℃ for 24 hours, whereby a concentration of 8X 10 was obtained 7 ～1×10 8 CFU/mL Lactobacillus salivarius fermentation broth.
(3) And (3) test development: the test was carried out for 7 days in 6 aquaria. 5L of clean seawater is filled into each jar, 65 shrimp fries are stocked in each jar, and 390 shrimp larvae of litopenaeus vannamei are bred in total. The specific process is as follows:
the test was divided into 2 groups, 1 being a blank control group and 1 being a test group, each group being set 3 replicates. The volume of the aquarium used for the test is 8L, the aquarium is disinfected by 0.01M potassium permanganate solution before irrigation, then is washed by sterile water for 3 times, and then is irrigated by 5L of clean seawater with the salinity of 15 per mill in each barrel. Before the culture test, the water is aerated, and the oxygen dissolving amount of the culture water is maintained to be more than 5ppm by adopting an air pump. During the test, the indoor temperature was maintained at about 28 ℃.
390 shrimp larvae were randomly divided into 2 groups of 3 replicates each containing 65 shrimp larvae. All shrimp larvae are cultured in an aquarium for 24 hours before experiment implementation so as to adapt to the environment. In a test group, the lactobacillus salivarius fermentation liquor is mixed with the feed, stands for 10min and then is fed into shrimp larvae, and the final concentration in the water body is 1 multiplied by 10 5 CFU/mL. In the blank control group, no lactobacillus salivarius fermentation broth was added. Basic bait is fed to each group of shrimp larvae, the basic bait is special feed shrimp slices for the shrimp larvae provided by a shrimp larvae incubator farm in Guangdong province, the feeding amount of the basic bait is 0.5mg of the basic bait per 10 shrimp larvae, the basic bait is fed every 8 hours every day, and residual bait is removed periodically.
The cultivation adopts a unified management mode, the health condition of the shrimp larvae is continuously observed for 7 days, and the body length and the weight of the shrimp larvae are respectively measured on the 0 th day and the 7 th day. And after the test is finished, recording the number of the live shrimps and calculating the survival rate.
(4) And (3) test results:
TABLE 1 growth Performance index between blank control and test groups
| Reference index | Blank control group | Test group |
| Length of 0 th day body | 11.17±0.17 a | 10.17±0.25 b |
| Length of day 7 | 10.60±0.24 b | 11.27±0.50 a |
| Body length growth (TLG) | -0.57±0.11 b | 1.10±0.72 a |
| Body length growth rate (PTLG) | -5.08±1.11 b | 10.90±7.40 a |
| Body weight of |
0.92±0.02 a | 0.75±0.12 b |
| Body weight on day 7 | 1.00±0.09 a | 1.04±0.06 a |
| Weight Gain (TWG) | 0.08±0.08 b | 0.29±0.18 a |
| Rate of body weight gain (PTWG) | 8.13±5.03 b | 43.48±19.76 a |
| Specific Growth Rate (SGR) | 1.16±0.72 b | 6.20±2.82 a |
| Survival Rate (SR) | 89.67±5.90 a | 89.67±2.49 a |
Note: SR (%) ═ 100 xn 7 /N 0
TLG(mm)＝L 7 -L 0
PTLG(％)＝100×(L 7 -L 0 )/L 0
TWG(mg)＝W 7 -W 0
PTWG(％)＝100×(W 7 -W 0 )/W 0
SGR(mg/day)＝100×(W 7 -W 0 )/7
Wherein N is 7 The sum of the healthy shrimp larvae after 7 days of test and the tail number of the sampled shrimp larvae in the test process; n is a radical of 0 The initial shrimp fry number is obtained; l is 7 Final average body length; l is 0 Is the initial average body length; w 7 Final average body weight; w 0 Mean initial body weight.
The difference of the upper-marked letters of the numerical values of the same row represents obvious difference, and p is less than 0.05; the converse indicates no significant difference, p > 0.05.
As can be seen from Table 1, the survival rate of the shrimp larvae in the test group was equivalent to that in the blank control group (p > 0.05).
TWG, PTWG and SGR were significantly higher in the test group than in the blank control group (p <0.05) in terms of weight gain; both TLG and PTLG were significantly higher in the test group than the blank control group (p <0.05) in terms of growth. It can be seen that the test group shrimp larvae showed significant growth advantages.
In conclusion, the lactobacillus salivarius fermentation broth can effectively improve the growth performance of the young litopenaeus vannamei (PL7-8), promote the growth of the body and the weight, and improve the economic benefit for the production and breeding industry of the young litopenaeus vannamei.
Example 2 Metabonomics study of Lactobacillus salivarius fermentation broth on Litopenaeus vannamei larvae (PL7-8)
Based on example 1, taking shrimp larvae at day 7 to perform LC-MS non-targeted metabonomics analysis, and the experimental process is as follows:
(1) sample pretreatment: accurately weighing 50mg of prawn sample, and dropwise adding 400 mu L of cold methanol solution (methanol: water-4: 1); crushing at 4 deg.C in a high throughput tissue crusher; vortex and mix well, extract ultrasonically for 10min on ice, repeat 3 times. The sample was then left to stand at-20 ℃ for 30 min. The cells were then centrifuged at 13000g at 4 ℃ for 15min, and after centrifugation, the supernatant was transferred to LC-MS liner injection vials for in-flight analysis. In order to evaluate the stability of the analytical system during the operation of the apparatus, a Quality Control sample (Quality Control, QC) is prepared. QC samples were prepared by mixing all test samples. During the analysis process of the instrument, one QC sample is inserted into every 6-8 analysis samples. During data analysis, the aggregation degree of the QC samples can be used for evaluating the stability of the instrument in the whole analysis process, and can also be used for capturing variable with large variation in an analysis system to ensure the reliability of results.
(3) LC-MS analysis: and (3) chromatographic separation: mobile phase a was water (containing 0.1% formic acid), mobile phase B was acetonitrile/isopropanol (1: 1) (containing 0.1% formic acid); the flow rate was 0.40mL/min, the amount of sample was 10. mu.L, and the column temperature was 40 ℃. Mass spectrum collection: detection was performed using ion spray voltage (ESI) as the ion source for mass spectrometry. Mass spectrum source parameters: the mass range is as follows: 70-1050; sheath gas: 40 psi; auxiliary heating gas: 10 psi; ion source heating temperature: 400 ℃; ionization voltage (positive electrode): + 3500V; ionization voltage (negative electrode): 2800V. Collision energy: 20-40-60V.
(4) Data preprocessing and library searching and identification: after the machine is completed, the LC-MS raw data is converted into a format and then introduced into prognesis QI (Waters Corporation, Milford, USA) for data preprocessing, and the procedures include baseline filtration, peak identification, peak area automatic integration, retention time correction, peak alignment, and the like. The MS and MS/MS mass spectral information is then matched to public databases (e.g., HMDB and Metlin) by Progenesis QI software library identification. Metabolites are identified by a mass to charge ratio (m/z), mass number and retention time in general. Finally, a data matrix with retention time, mass-to-charge ratio, adduct and other parameters is obtained.
(5) Statistical analysis: before statistical analysis, the data matrix is normalized by total peak area, and the variable of QC sample Relative Standard Deviation (RSD) is deleted, wherein the variable is more than or equal to 30%. Sample Principal Component Analysis (PCA) and orthogonal least squares discriminant analysis (OPLS-DA) were performed using the R software package ropls tool (version 1.6.2), and 200 permutation tests were performed to assess the accuracy of the model.
4. Differential metabolite screening
And screening the metabolites with significant difference among groups by taking VIP (variable weight value) >1 and p <0.05 as thresholds. Meanwhile, the metabolites provided with the KEGG ID numbers are mapped to pathway paths for enrichment analysis, and the biological process that lactobacillus salivarius affects the growth of shrimp larvae is deeply excavated.
5. Results of the experiment
(1) Differential metabolite screening and hierarchical clustering analysis
There were 33 significantly different metabolites that met the screening threshold. As shown in fig. 1 and table 2, the test group had 25 up-regulated metabolites and 8 down-regulated metabolites relative to the blank control group. Wherein the significantly different metabolites include sphingolipids, lipid acyl compounds, amino acids, short peptides and derivatives thereof, pregnenolone lipids, steroids and derivatives thereof, benzene compounds, flavonoids and vitamins.
a. Amino acids, short peptides and derivatives thereof: the short peptide plays an important role in improving the oxidation resistance of organisms. On the 7 th day, the compounds are remarkably up-regulated in a test group (p is less than 0.05), which shows that lactobacillus salivarius has a promoting effect on the synthesis of short peptides, improves the oxidation resistance of organisms and protects PUFAs from lipid peroxidation.
b. Steroids and their derivatives: steroids and derivatives thereof belong to isoprene substances, and the compounds are closely related to crustacean molting and growth regulation. On day 7, the steroid and its derivatives were significantly up-regulated in the L group (p <0.05), indicating that lactobacillus salivarius could promote the production of shrimp larvae steroid and its derivatives, and further shorten the molting cycle of prawns, which is consistent with the results of lactobacillus groups on increasing shrimp body length.
c. Benzene compounds: benzene compounds generally inhibit the production of superoxide and have good anti-inflammatory and antibacterial properties. Adipostatin A and 6-Gingerol contain phenolic groups in their structure, and the antioxidant properties of phenols may be related to their ability to donate electrons and act as radical scavengers by forming stable phenoxy radicals. Thus, upregulation of benzene compounds is a metabolic manifestation of the maintenance of the balance of oxygen free Radicals (ROS) in the body. On day 7, the benzene compounds were significantly up-regulated in group L (p <0.05), indicating that Lactobacillus salivarius has an effect of promoting the synthesis of benzene compounds.
d. Flavonoid: flavonoids have potential biological activities including direct antibacterial activity, synergy with antibiotics, and inhibition of bacterial virulence. On day 7, the content of flavonoids in group L was significantly higher than that in group C (p <0.05), indicating that Lactobacillus salivarius has a promoting effect on flavonoid synthesis.
TABLE 2 metabolites significantly different between test group and blank control group
Note: "VIP" refers to variable weight values, VIP >1 representing that the metabolite is able to distinguish between samples. The greater the VIP value, the more significant the contribution of the metabolite to the inter-group differences. "pvalue" was tested by independent sample T to verify whether the metabolites in the test group and the blank group were significant, pvalue <0.05 represents that the metabolites were significantly different between the two groups. "regulated" represents the variation of group L metabolites relative to group C metabolites.
(2) KEGG pathway enrichment assay
KEGG pathway enrichment analysis is shown in figure 2 and table 3. The results showed that there were 6 significantly enriched metabolic pathways (p <0.05) of Terphenonoid biosynthesis (biosynthesis of terpene skeleton), Vitamin differentiation and adsorption (digestion and absorption of vitamins), beta-Alanine metabolism (β -Alanine metabolism), pantotenoate and CoA biosynthesis (biosynthesis of pantothenic acid and CoA), Sphingolipid metabolism (Sphingolipid metabolism) and Sphingolipid signaling pathway (Sphingolipid signaling pathway), respectively. Among the enriched metabolites are Farnesylcysteine, Panthothinic Acid and Sphinganine, respectively.
Farnesyl cysteine belongs to sesquiterpenes and is a key endogenous component capable of promoting molting of crustaceans. In the biosynthesis pathway of the terpene skeleton, farnesyl cysteine is up-regulated in the pathway, which indicates that lactobacillus salivarius can activate the pathway and promote exuviation of shrimp larvae, and the result is consistent with the result that the body length yield of L groups of shrimp larvae is obviously increased.
Pantothenic acid, also known as vitamin B5, is synthesized by gut commensal bacteria in the absence of dietary supplements. In the pathway of pantothenate and CoA biosynthesis, pantothenate forms acetyl CoA under the combined action of an enzyme system consisting of pantothenate kinase, phosphopantothenate-cysteine ligase, PPC-decarboxylase, dephosphoroyl-CoA pyrophosphorylase, and dephosphorylated creatine kinase. Acetyl CoA is a key metabolic intermediate in carbon catabolic pathways and energy metabolism, including glycolysis, pyruvate oxidation, and fatty acid beta oxidation. Therefore, the application of the lactobacillus salivarius is considered to be helpful for regulating the biosynthesis of pantothenic acid and CoA in the shrimp larvae body, promoting the synthesis of acetyl CoA, participating in energy metabolism and providing enough energy for the body to grow.
In summary, from the metabolic aspect, lactobacillus salivarius can regulate the growth traits of the organism by regulating up-regulation/down-regulation of metabolites.
Table 3 KEGG pathway metabolite enrichment profile
| KEGG pathway | Metabolites |
| Terpenoid backbone biosynthesis* | Farnesylcysteine(↑) |
| Vitamin digestion and absorption* | Pantothenic Acid(↑) |
| beta-Alanine metabolism* | Pantothenic Acid(↑) |
| Pantothenate and CoA biosynthesis* | Pantothenic Acid(↑) |
| Sphingolipid metabolism* | Sphinganine(↑) |
| Sphingolipid signaling pathway* | Sphinganine(↑) |
Note: "x" indicates significant enrichment of the pathway, and "meshed" indicates upregulation of levels in the experimental group relative to the blank control group.
The above embodiments are preferred embodiments of the present invention, but the present invention is not limited to the above embodiments, and any other changes, modifications, substitutions, combinations, and simplifications which do not depart from the spirit and principle of the present invention should be construed as equivalents thereof, and all such changes, modifications, substitutions, combinations, and simplifications are intended to be included in the scope of the present invention.
Claims (10)
1. The application of the lactobacillus salivarius in the cultivation of the young shrimps of the litopenaeus vannamei is characterized in that:
the Lactobacillus salivarius is Lactobacillus salivarius GZPH 2.
2. Use of a Lactobacillus salivarius as claimed in claim 1 to improve the growth performance of Litopenaeus vannamei.
3. Use according to claim 2, characterized in that:
the lactobacillus salivarius is applied to improving the growth performance of the young litopenaeus vannamei.
4. Use of lactobacillus salivarius as claimed in claim 1 in promoting ecdysis and/or growth-related synthesis of secondary metabolites of young litopenaeus vannamei.
5. Use according to claim 4, characterized in that:
the lactobacillus salivarius is used for up-regulating metabolite short peptides and amino acid derivatives of litopenaeus vannamei larvae, sesquiterpenes, steroids and derivatives thereof, benzene compounds, flavonoids and/or pantothenic acid.
6. Use according to claim 5, characterized in that:
the short peptide is at least one of Methionyl-Proline, Valyl-Proline, Tyrosyl-Proline and L-phenylallyl-L-Proline;
the amino acid derivative is Glutamate, gamma-methyl ester;
the sesquiterpene is farnesyl cysteine;
the steroid and the derivative thereof are at least one of 3a,21-Dihydroxy-5b-pregnane-11,20-dione, 3 b-allotetrahydrocortisone, Convalloside and 2-deoxyastasterone;
the benzene compound is at least one of N- [ (4-Hydroxy-3-methoxyphenyl) methyl ] octanamide, 6-Gingerol and Adipostatin A;
the flavonoid is Formononetin.
7. Use according to any one of claims 1 to 6, characterized in that:
the application is realized by mixing lactobacillus salivarius fermentation liquor and feeding the mixed liquor into a litopenaeus vannamei fry culture water body.
8. Use according to claim 7, characterized in that:
the concentration of Lactobacillus salivarius in water is 10 4 ～10 6 CFU/mL。
9. Use according to claim 7, characterized in that:
the preparation method of the lactobacillus salivarius fermentation liquor comprises the following steps:
inoculating single lactobacillus salivarius colony in MRS liquid culture medium, and culturing to obtain seed liquid; then inoculating the seed liquid into an MRS liquid culture medium, and culturing again to obtain the lactobacillus salivarius fermentation liquid.
10. Use according to claim 9, characterized in that:
the inoculation amount of the seed liquid is 1-5% v/v;
the culture condition is that the culture is carried out for 12-36 h under the conditions of 16-40 ℃ and 200 rev/min;
the re-culture condition is to culture for 12-36 hours at 16-40 ℃ under the condition of standing.
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| CN116103208A (en) * | 2023-04-07 | 2023-05-12 | 西南科技大学 | Application of lactobacillus salivarius in antioxidation |
| CN116784421A (en) * | 2023-06-21 | 2023-09-22 | 华南理工大学 | Application of lactobacillus salivarius in bullfrog tadpole culture |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN116103208A (en) * | 2023-04-07 | 2023-05-12 | 西南科技大学 | Application of lactobacillus salivarius in antioxidation |
| CN116103208B (en) * | 2023-04-07 | 2024-01-16 | 西南科技大学 | Application of lactobacillus salivarius in antioxidation |
| CN116784421A (en) * | 2023-06-21 | 2023-09-22 | 华南理工大学 | Application of lactobacillus salivarius in bullfrog tadpole culture |
| CN116784421B (en) * | 2023-06-21 | 2024-05-31 | 华南理工大学 | Application of lactobacillus salivarius in bullfrog tadpole culture |
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