CN103947886A - Formula and preparation method of large yellow croaker lactobacillus plantarum feed - Google Patents

Formula and preparation method of large yellow croaker lactobacillus plantarum feed Download PDF

Info

Publication number
CN103947886A
CN103947886A CN201410188158.6A CN201410188158A CN103947886A CN 103947886 A CN103947886 A CN 103947886A CN 201410188158 A CN201410188158 A CN 201410188158A CN 103947886 A CN103947886 A CN 103947886A
Authority
CN
China
Prior art keywords
lactobacillus plantarum
large yellow
feed
yellow croaker
fish
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
CN201410188158.6A
Other languages
Chinese (zh)
Inventor
廖志勇
缪雄伟
林利
李小花
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Wenzhou University
Original Assignee
Wenzhou University
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Wenzhou University filed Critical Wenzhou University
Priority to CN201410188158.6A priority Critical patent/CN103947886A/en
Publication of CN103947886A publication Critical patent/CN103947886A/en
Pending legal-status Critical Current

Links

Classifications

    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
    • Y02A40/80—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in fisheries management
    • Y02A40/81—Aquaculture, e.g. of fish
    • Y02A40/818—Alternative feeds for fish, e.g. in aquacultures
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
    • Y02P60/00—Technologies relating to agriculture, livestock or agroalimentary industries
    • Y02P60/80—Food processing, e.g. use of renewable energies or variable speed drives in handling, conveying or stacking
    • Y02P60/87—Re-use of by-products of food processing for fodder production

Abstract

The invention discloses a formula and a preparation method of large yellow croaker lactobacillus plantarum feed. The formula of the large yellow croaker lactobacillus plantarum feed comprises feeding feed and basal feed, wherein the feeding feed comprises 1*10<6> cfu/kg of lactobacillus plantarum, 0.1g/kg of thallus, 9.999g/kg of milk, 540g/kg fish meal, 183g/kg of starch, 37g/kg of fish paste, 20g/kg of gluten, 20g/kg of fish oil, 64g/kg of shrimp shell powder, 94g/kg of defatted soybean meal, 35g/kg of mineral mixture and 7g/kg of vitamin mixture, the percentage by mass of the feeding feed is 25.2%, and the basal feed comprises 47.3% of crude protein, 8.9% of crude fat, 10.2% of ash content and 8.4% of moisture content in percentage by mass. The preparation method comprises the following steps: carrying out separation culture, collecting the lactobacillus plantarum, mixing the defatted milk, adding the basal feed, mixing and preparing. The large yellow croaker lactobacillus plantarum feed can be used for increasing growth rate of body mass of large yellow croakers and feeding rate of the large yellow croakers, improving non-specific immunity performances such as complement activity, lysozyme activity, phagocytic activity and respiration of leukocytes in head and kidney of the large yellow croakers and enhancing pathogenic bacteria resistance and facilitates development of large yellow croaker breeding industries.

Description

A kind of large yellow croaker Lactobacillus plantarum feed formula and preparation method thereof
Technical field
The invention belongs to large yellow croaker cultural technique field, relate in particular to a kind of large yellow croaker Lactobacillus plantarum feed formula and preparation method thereof.
Background technology
Large yellow croaker is the fish that China is exclusive, economic worth is very high, is one of kind the famousst and precious in China's marine fish culture.In recent years, the cultivation scale of large yellow croaker expands rapidly due to the success of artificial breeding, and due to overfishing, natural resources is day by day exhausted.After artificial breeding is obtained success, the net cage scale of large yellow croaker expands rapidly.The unreasonable planning of cage culture causes cultured area and breeding density sharply to strengthen, make the bearing capacity and the water body exchange self-purification capacity that cultivate waters suffer serious harm, the kind of fish diseases also increases thereupon, is greatly restricting the sound development of large yellow croaker aquaculture.Along with cultured area and breeding density sharply strengthen, breeding environment is destroyed, and the kind of disease also increases thereupon, morbidity number of times is increased, harmfulness increases rapidly, is faced with many problems relevant with nutrient fodder, is greatly restricting the sound development of large yellow croaker aquaculture.For example, large yellow croaker cultivation is normal to be occurred and viral relevant with bacterium epidemic situation, causes serious economic loss.Because the feed coefficient of current use is not up to standard, the probability that is converted into fish protein is low, causes bait to run off in a large number, and water quality is caused to severe contamination simultaneously.Especially part raiser Misuse forbidding fish medicine and feed inferior, cause medicament residue and water quality large area to worsen, cause that the large yellow croaker speed of growth slows down, disease takes place frequently, survival rate declines, and quality of large yellow croaker declines, food security do not ensure, the health of fish and the raising of its immunity are just particularly important for aquaculture.Probio is the microbiological feed adding ingredient that a class produces beneficial effect by improving host animal intestinal microecology to it, host animal health is existed to the microorganism of beneficial effect.Probio series products mostly has raising breeding performonce fo animals to animal, improves health, and improves intestinal microecology, improves the effects such as immunity.
At present, traditional large yellow croaker feed exists the speed of growth slow, and sick rate is high, immunity and poor quality, and survival rate is low, the problem that food security does not ensure.
Summary of the invention
The object of the embodiment of the present invention is to provide a kind of large yellow croaker Lactobacillus plantarum feed formula and preparation method thereof, is intended to solve at present, and traditional large yellow croaker feed exists the speed of growth slow, sick rate is high, immunity and poor quality, survival rate is low, the problem that food security does not ensure.
The embodiment of the present invention is achieved in that a kind of large yellow croaker Lactobacillus plantarum feed formula, and this large yellow croaker Lactobacillus plantarum feed formula comprises: Lactobacillus plantarum 1 × 10 6the basal feed of the feed for nursing of cfu/kg, thalline 0.1g/kg, milk 9.999g/kg, fish meal 540g/kg, starch 183g/kg, fish cream 37g/kg, gluten 20g/kg, fish oil 20g/kg, shrimp shell meal 64g/kg, defatted soybean meal 94g/kg, mineral intermixture 35g/kg, vitamin mixtures 7g/kg composition and crude protein, thick fat, ash content and water composition;
According to mass percent, feed for nursing is 25.2%, crude protein 47.3%, thick fat 8.9%, ash content 10.2%, moisture 8.4%.
Further, this large yellow croaker Lactobacillus plantarum feed formula is: Lactobacillus plantarum 1 × 10 8cfu/kg, thalline 0.1g/kg, milk 9.99g/kg, fish meal 540g/kg, starch 183g/kg, fish cream 37g/kg, gluten 20g/kg, fish oil 20g/kg, shrimp shell meal 64g/kg, defatted soybean meal 94g/kg, mineral intermixture 35g/kg, vitamin mixtures 7g/kg, crude protein 47.3%, thick fat 8.9%, ash content 10.2%, moisture 8.4%.
Further, this large yellow croaker Lactobacillus plantarum feed formula is: Lactobacillus plantarum 1 × 10 10cfu/kg, thalline 10g/kg, fish meal 540g/kg, starch 183g/kg, fish cream 37g/kg, gluten 20g/kg, fish oil 20g/kg, shrimp shell meal 64g/kg, defatted soybean meal 94g/kg, mineral intermixture 35g/kg, vitamin mixtures 7g/kg, crude protein 47.3%, thick fat 8.9%, ash content 10.2%, moisture 8.4%.
Another object of the embodiment of the present invention is to provide a kind of preparation method of large yellow croaker Lactobacillus plantarum feed, and the preparation method of this large yellow croaker Lactobacillus plantarum feed comprises the following steps:
Separate and obtain Lactobacillus plantarum from Korea S's pickled cabbage, in the sterile flask that 37 DEG C are incubated at containing MRS meat soup, after 24 hours, centrifugal 5 minutes of 15400 × g, collects Lactobacillus plantarum mixture;
Mix with skim milk in 1:4 ratio, freeze drying, by the viability of MRS agar plate numeration mensuration Lactobacillus plantarum mixture;
According to Lactobacillus plantarum 1 × 10 6cfu/kg, thalline 0.001g/kg, milk 9.999g/kg, fish meal 540g/kg, starch 183g/kg, fish cream 37g/kg, gluten 20g/kg, fish oil 20g/kg, shrimp shell meal 64g/kg, defatted soybean meal 94g/kg, mineral intermixture 35g/kg, vitamin mixtures 7g/kg configuration large yellow croaker feed for nursing;
Adding composition is the basal feed of 47.3% crude protein, 8.9% thick fat, 10.2% ash content and 8.4% moisture, is mixed and made into large yellow croaker Lactobacillus plantarum feed.
Further, this large yellow croaker Lactobacillus plantarum feed is fed large yellow croaker twice in one day.
Large yellow croaker Lactobacillus plantarum feed formula provided by the invention and preparation method thereof, adopting content is 10 6-10 10the formula feed of cfu/kg Lactobacillus plantarum, body weight gain rate and the grazing rate of large yellow croaker are promoted, and strengthen the resistance to germ Streptococcussp, promote complement activity, lysozyme activity, activate the phagocytic capacity and the head-kidney leucocyte respiratory of large yellow croaker nospecific immunity performance.The present invention is conducive to the aquaculture development of large yellow croaker.The preparation method of the Lactobacillus plantarum feed of large yellow croaker is simple, large yellow croaker fast growth, and immunity is strong, and sick low with the death rate, edible safety is good.
Brief description of the drawings
Fig. 1 is preparation method's flow chart of the large yellow croaker Lactobacillus plantarum feed that provides of the embodiment of the present invention;
Fig. 2 is that the large yellow croaker feeding that the embodiment of the present invention provides contains 0,1 × 10 4, 1 × 10 6, 1 × 10 8with 1 × 10 10the body weight gain rate schematic diagram in cfu/kg Lactobacillus plantarum feed 1-4 week;
Fig. 3 is that the large yellow croaker feeding that the embodiment of the present invention provides contains 0,1 × 10 4, 1 × 10 6, 1 × 10 8with 1 × 10 10the feeding efficiency schematic diagram in cfu/kg Lactobacillus plantarum feed 1-4 week;
Fig. 4 is PCR qualification schematic diagram after feed surrounding in the large yellow croaker feeding table one that provides of the embodiment of the present invention;
Fig. 5 is the complement activity (ACH of the large yellow croaker feeding that provides of the embodiment of the present invention containing Lactobacillus plantarum feed 1-4 week 50) schematic diagram;
Fig. 6 is that the large yellow croaker feeding that the embodiment of the present invention provides breaks out breathing power schematic diagram containing Lactobacillus plantarum, the head-kidney leucocyte in feed 1-4 week;
Fig. 7 is the head-kidney leukocytes phagocytic active schematic diagram of the large yellow croaker feeding that provides of the embodiment of the present invention containing Lactobacillus plantarum feed 1-4 week;
Fig. 8 is serum lysozyme activities reached (ACH50) schematic diagram of the yellow croaker feeding that provides of the embodiment of the present invention containing Lactobacillus plantarum feed 1-4 week.
Detailed description of the invention
In order to make object of the present invention, technical scheme and advantage clearer, below in conjunction with embodiment, the present invention is further elaborated.Should be appreciated that specific embodiment described herein, only in order to explain the present invention, is not intended to limit the present invention.
Below in conjunction with drawings and the specific embodiments, application principle of the present invention is further described.
The large yellow croaker Lactobacillus plantarum feed formula of the embodiment of the present invention: Lactobacillus plantarum 1 × 10 6the basal feed of the feed for nursing of cfu/kg, thalline 0.1g/kg, milk 9.999g/kg, fish meal 540g/kg, starch 183g/kg, fish cream 37g/kg, gluten 20g/kg, fish oil 20g/kg, shrimp shell meal 64g/kg, defatted soybean meal 94g/kg, mineral intermixture 35g/kg, vitamin mixtures 7g/kg composition and crude protein, thick fat, ash content and water composition;
According to mass percent, feed for nursing is 25.2%, crude protein 47.3%, thick fat 8.9%, ash content 10.2%, moisture 8.4%.
Formula of the present invention can also be: Lactobacillus plantarum 1 × 10 8cfu/kg, thalline 0.1g/kg, milk 9.999g/kg, fish meal 540g/kg, starch 183g/kg, fish cream 37g/kg, gluten 20g/kg, fish oil 20g/kg, shrimp shell meal 64g/kg, defatted soybean meal 94g/kg, mineral intermixture 35g/kg, vitamin mixtures 7g/kg, crude protein 47.3%, thick fat 8.9%, ash content 10.2%, moisture 8.4%;
Formula of the present invention can also be: Lactobacillus plantarum 1 × 10 10cfu/kg, thalline 10g/kg, fish meal 540g/kg, starch 183g/kg, fish cream 37g/kg, gluten 20g/kg, fish oil 20g/kg, shrimp shell meal 64g/kg, defatted soybean meal 94g/kg, mineral intermixture 35g/kg, vitamin mixtures 7g/kg, crude protein 47.3%, thick fat 8.9%, ash content 10.2%, moisture 8.4%;
As shown in Figure 1, the preparation method of the large yellow croaker Lactobacillus plantarum feed of the embodiment of the present invention comprises the following steps:
S101: separate and obtain Lactobacillus plantarum from Korea S's pickled cabbage, in the sterile flask that 37 DEG C are incubated at containing MRS meat soup, after 24 hours, centrifugal 5 minutes of 15400 × g, collects Lactobacillus plantarum mixture;
S102: mix with skim milk in 1:4 ratio, freeze drying, by the viability of MRS agar plate numeration mensuration Lactobacillus plantarum mixture;
S103: according to Lactobacillus plantarum 1 × 10 6cfu/kg, thalline 0.001g/kg, milk 9.999g/kg, fish meal 540g/kg, starch 183g/kg, fish cream 37g/kg, gluten 20g/kg, fish oil 20g/kg, shrimp shell meal 64g/kg, defatted soybean meal 94g/kg, mineral intermixture 35g/kg, vitamin mixtures 7g/kg configuration large yellow croaker feed for nursing;
S104: adding composition is the basal feed of 47.3% crude protein, 8.9% thick fat, 10.2% ash content and 8.4% moisture, is mixed and made into large yellow croaker Lactobacillus plantarum feed.
In conjunction with following specific embodiment, result of use of the present invention is described further:
Embodiment 1:
1 materials and methods:
The preparation of 1.1 Lactobacillus plantarum mixture feeds:
Lactobacillus plantarum SaccharomycescerevisiaeP13 separates and obtains from Korea S's pickled cabbage [2], in the sterile flask that 37 DEG C are incubated at containing MRS meat soup, after 24 hours, 15, centrifugal 5 minutes of 400 × g, collects Lactobacillus plantarum mixture, mixes with skim milk in 1:4 ratio, freeze drying, measure the viability of Lactobacillus plantarum mixture by the numeration of MRS agar plate, as shown in table 1, according to 5 kinds of large yellow croaker feed for nursing of Lactobacillus plantarum content configuration, wherein basal feed composition is 47.3% crude protein, 8.9% thick fat, 10.2% ash content and 8.4% moisture, 1.0 × 10 9the Lactobacillus plantarum mixture of cfu/g joins in basal feed in the ratio shown in table 1, makes the content of Saccharomyces cerevisiaeP13 Lactobacillus plantarum in large yellow croaker feed for nursing reach respectively 0,10 4, 10 6, 10 8, 10 10cfu/kg;
1.2 large yellow croakers are raised:
Large yellow croaker juvenile fish is propagated field artificially purchased from local one, adopt water vat flowing water (seawater) feeding manner at the indoor raising experiment that carries out, in experimentation, change the seawater of 25-35% every day to ensure the quality of water, water temperature remains on 27.0 ± 0.5 DEG C, the pH value of water is 7.4-8.5, the salinity of water remains on 3.0 ± 0.2%, all experiment fishes are fed twice with the feed (table 1) of filling a prescription containing Lactobacillus plantarum SaccharomycescerevisiaeP13 every day, three groups of experiment fishes of the parallel nursing of each formula feed, totally 15 groups, every group of 5 large yellow croaker juvenile fish;
The mensuration of 1.3 large yellow croaker growths:
Large yellow croaker seedling is divided into 15 groups at random, every group rises in volume glass aquarium and feeds by the feed that contains Lactobacillus plantarum SaccharomycescerevisiaeP13 or control feed in 40-60, all aquariums are equipped with air cleaner, in growth measurement process, large yellow croaker feeding every day is containing Lactobacillus plantarum 0,10 4, 10 6, 10 8with 10 10cfu/kg feed, per week weighing once until determination experiment finishes, the computing formula that body weight increases percentage (PWG) and feed efficiency (FE) is as follows: PWG=[100% × (final weight-initial body weight)/(initial body weight)] and FE=(final weight-initial body weight)/(food ration);
1.4 large yellow croaker opposing germ bodies are analyzed:
Large yellow croaker feeding is containing 0,10 4, 10 6, 10 8with 10 10after the feed surrounding of cfu/kg Lactobacillus plantarum Saccharomyces cerevisiaeP13, the Streptococcussp. thalline of intramuscular injection respective amount stores night (5.0 × 10 respectively 10cfu/ml), so that the concentration of the interior initial Streptococcussp. of each experiment fish body is 5.0 × 10 5cfu/ gram of fish body weight, all experiment fishes all continue feeding containing 0,10 4, 10 6, 10 8with 10 10the feed of cfu/kg Lactobacillus plantarum SaccharomycescerevisiaeP13, adds up dead fish number every day;
The survival analysis of SaccharomycescerevisiaeP13 in 1.5 large yellow croaker enteron aisles:
The experiment of large yellow croaker feeding is with after formula feed surrounding, the fresh intestines of clip segment, pulverize with 20ml meat soup immediately, 37 DEG C of incubated overnight, 4 DEG C, 10, centrifugal 20 minutes of 000 × g, collecting precipitation, use the genome extraction agent box separation and purification SaccharomycescerevisiaeP13 genomic DNA purchased from the green skies, on Perkin-Elmer9700 nucleic acid augmentative instrument with special primer pair 5 '-TCTGTATATTCTGTATCTATGTTCCTGC-3 ' and 5 '-AAATGGCCTATTGTATTGTCAGGTC-3 ' ABI company of SaccharomycescerevisiaeP13, carry out PCR (PCR) amplification, first 94 DEG C of heat denatured 10 minutes, follow 95 DEG C of sex change 30 seconds, anneal 30 seconds for 55 DEG C, 72 DEG C are extended 45 seconds, 35 circulations, last 72 DEG C are extended 10 minutes, PCR product separates qualification by 2.0% agarose electrophoresis,
2 results and analysis:
The impact of 2.1SaccharomycescerevisiaeP13 on large yellow croaker body weight gain rate:
Feeding containing the large yellow croaker after the formula feed (table 1) of Lactobacillus plantarum SaccharomycescerevisiaeP13 from first week, body weight gain rate is just apparently higher than the large yellow croaker body weight gain rate of feeding basal feed (control feed), after surrounding, feeding control feed and containing 1 × 10 4, 1 × 10 6, 1 × 10 8with 1 × 10 10the large yellow croaker body weight gain rate of cfu/kg Lactobacillus plantarum SaccharomycescerevisiaeP13 is respectively 88.9 ± 3.1%, 101.1 ± 5.8%, 132.4 ± 11%, 140.8 ± 6.7% and 138.9 ± 6.4% (Fig. 2); In Fig. 2, large yellow croaker feeding is containing 0,1 × 10 4, 1 × 10 6, 1 × 10 8with 1 × 10 10the body weight gain rate in cfu/kg Lactobacillus plantarum SaccharomycescerevisiaeP13 feed 1-4 week.The mean value that each numerical value equals every group of 3 measured values adds standard error, p ﹤ 0.05;
Table 1 large yellow croaker feed for nursing composition
2.2SaccharomycescerevisiaeP13 improves the feeding efficiency of large yellow croaker:
The raising of large yellow croaker body weight gain rate may come from the increase of food ration on the one hand, and may come from the other hand the raising of feeding efficiency, and finally ascribe the raising of large yellow croaker body internal protein conversion ratio to, as shown in Figure 2, feeding control feed and containing 1 × 10 4, 1 × 10 6, 1 × 10 8with 1 × 10 10the feeding efficiency of the large yellow croaker of cfu/kg Lactobacillus plantarum SaccharomycescerevisiaeP13 feed is respectively 0.81 ± 0.23,0.98 ± 0.15,1.10 ± 0.12,1.29 ± 0.24 and 1.31 ± 0.26, the feeding efficiency that large yellow croaker is described improves with the increase of Lactobacillus plantarum SaccharomycescerevisiaeP13 content in feed, thereby improves the efficiency of actual of feed; Fig. 3 large yellow croaker feeding is containing 0,1 × 10 4, 1 × 10 6, 1 × 10 8with 1 × 10 10the feeding efficiency in cfu/kg Lactobacillus plantarum SaccharomycescerevisiaeP13 feed 1-4 week, the mean value that each numerical value equals every group of 3 measured values adds standard error, p ﹤ 0.05;
2.3SaccharomycescerevisiaeP13 improve large yellow croaker infect. after survival rate:
Lactobacillus plantarum can strengthen the resistance that grouper infects Streptococcussp., has analyzed the resistance that feeding infects common disease thalline containing the large yellow croaker of Lactobacillus plantarum SaccharomycescerevisiaeP13, feeding control feed and containing 1 × 10 4, 1 × 10 6, 1 × 10 8with 1 × 10 10the large yellow croaker of cfu/kg Lactobacillus plantarum SaccharomycescerevisiaeP13 is respectively 35.7 ± 6.7%, 80.0 ± 5.8%, 76.7 ± 3.3%, 70.0 ± 5.8% and 70.0 ± 5.8% (table 2) in the survival rate being subject to after germ body Streptococcussp. infection surrounding, the growth that is not only conducive to large yellow croaker containing the feed of Lactobacillus plantarum SaccharomycescerevisiaeP13 is described, is also conducive to the resistance of large yellow croaker to courses of infection simultaneously;
Table 2 large yellow croaker feeding infects the survival rate after Streptococcus sp. containing the large yellow croaker of Saccharomyces cerevisiae P13 feed
The detection of SaccharomycescerevisiaeP13 after 2.4 large yellow croaker feedings:
Large yellow croaker feeding is containing 0,1 × 10 4, 1 × 10 6, 1 × 10 8with 1 × 10 10after the feed surrounding of cfu/kg Lactobacillus plantarum SaccharomycescerevisiaeP13, by the PCR SaccharomycescerevisiaeP13 genetic fragment that can increase specifically, and the amount of PCR product increases (Fig. 4) with the raising of SaccharomycescerevisiaeP13 content in feed;
Probio is that a class produces the microbiological feed adding ingredient of beneficial effect by improving host animal intestinal microecology to animal growth and immunity, probio or probiotics fermention product can promote rainbow trout effectively, the growth of grouper etc., and strengthen fish to common disease thalline infect opposing, the present invention is by measuring the body weight gain rate of large yellow croaker, feeding efficiency and the resistance to encountered pathogenic bacteria, study the impact of probio SaccharomycescerevisiaeP13 on large yellow croaker growth, the growth rate of the large yellow croaker that result demonstration feeding contains SaccharomycescerevisiaeP13 feed, the utilization rate of feeding feed and the resistance of germ is all significantly improved, show, containing the biological feedstuff of SaccharomycescerevisiaeP13, large yellow croaker cultivation is had to larger potential value.
Embodiment 2:
1 materials and methods:
1.1 large yellow croakers are raised:
Large yellow croaker juvenile fish is propagated field artificially purchased from local one, carry out raising experiment in indoor employing water vat flowing water (seawater) feeding manner, in experimentation, change 30% seawater every day to ensure the quality of water, water temperature remains on 28 ± 0.2 DEG C, the pH value of water is 7.5-8.4, the salinity of water remains on 2.0 ± 0.1%, Lactobacillus plantarum SaccharomycescerevisiaeP13 separates and obtains from Korea S's pickled cabbage, in the sterile flask that 37 DEG C are incubated at containing MRS meat soup, after 24 hours, 15, centrifugal 5 minutes of 400 × g, collect Lactobacillus plantarum mixture, mix with skim milk in 1:4 ratio, freeze drying, measure the viability of Lactobacillus plantarum mixture by the numeration of MRS agar plate, all experiment fishes use formula feed or the basal feed prepared by table one to feed twice every day, three parallel group experiment fish of each formula forage feed, totally 15 groups, every group of 5 large yellow croaker juvenile fish, after the feed surrounding that large yellow croaker feeding contains lactic acid bacteria SaccharomycescerevisiaeP13, measure respectively related immune parameter,
1.2 complement activity (ACH 50) mensuration:
Measure complement activity, as target cell, use EGTA-Mg with sheep red blood cell 2+-GVB buffer solution dilutes the blood sample from large yellow croaker in proportion, respectively getting 20 μ l originates as complement, mix with 6 μ l sheep red blood cell suspension, 21 DEG C, pH7.2 is hatched 2 hours, add 200 μ l to stop hemolytic reaction containing the GVB buffer solution of EDTA, 4 DEG C, centrifugal 10 minutes of 1600 × g, survey the OD value of supernatant by enzyme linked immunosorbent assay (ELISA), in hemolytic reaction liquid, use respectively 6 μ lEDTA-GVB, 20 μ LEDTA-GVB change, 200 μ l deionized waters replace sheep red blood cell suspension, dilution large yellow croaker blood sample basis and diluted blood sample/EDTA-GVB, as SRBC, the blank of blood sample basis and 100% haemolysis sample, haemolysis degree calculates as follows:
1.3 antiplasmin activities are measured:
Serum antiplasmin activity is measured according to bibliographical information method [3]10 μ l serum samples mix with the Micrococcusluteus suspension that 200 μ l are dissolved in the PBS (0.05M) of pH6.2,27 DEG C of incubations 6 minutes, read plate with ELISA and survey its OD value at 530nm, and every milliliter of blood sample reduction per minute 0.001OD value is defined as a unit antiplasmin activity;
1.4 outburst respiration monitorings:
Large yellow croaker head-kidney leucocyte outburst respiratory activity method is measured, the pre-embedding micro plate of poly-L-lysine solution (100 μ l/ hole) with 0.20%, and every hole adds 100 μ l leukocyte suspensions (1 × 10 subsequently 6cells/ml), centrifugal 20 minutes of 700 × g, remove not adherent cell, clean micro-plate hole with HBSS liquid, every hole adds the HBSS liquid containing 0.1% zymosan, room temperature reaction 30 minutes, add separately position, the hole blank of 100 μ lHBSS liquid, wash cell 3 times with HBSS liquid, add 100 μ l0.3%NBT liquid, room temperature dyeing 30 minutes, remove NBT liquid, and add 100 μ l methyl alcohol to stop staining reaction, add 120 μ lKOH (2.0M) and 140 μ lDMSO, read plate instrument and measure the OD value of every porocyte lysate with ELISA, each mensuration is parallel to be done 3 times,
The mensuration of 1.5 activate the phagocytic capacity:
Add 500 μ l containing 1 × 10 toward every hole in 12 orifice plates 6the leukocytic L-15 liquid of individual head-kidney, 4 DEG C in 400 × g centrifugal 20 minutes, remove non-adherent cell, and clean cell with L-15 liquid, every hole adds 1 μ l containing 1 × 10 6the liquid of fluorescence latex bead, 28 DEG C of incubations 2 hours, wash cell 3 times with PBS liquid, to remove the fluorescence latex bead not being adsorbed, after fixing with 10% formalin, wash 3 times with PBS, with the 0.1% propidium iodide 10min that dyes, wash 3 times with PBS again, under fluorescence microscope, select 100 phagocytes to calculate the cytophagous percentage of picked-up perforated glass pearl;
2 results and analysis:
2.1 Lactobacillus plantarums improve large yellow croaker complement activity (ACH 50):
Feeding contains Lactobacillus plantarum (SaccharomycescerevisiaeP13) 1 × 10 6, 1 × 10 8with 1 × 10 10the large yellow croaker ACH of cfu/kg feed 50contain Lactobacillus plantarum 1 × 10 apparently higher than feeding 4or the large yellow croaker ACH of control feed 50, than the ACH of control group large yellow croaker 50exceed respectively 32 ± 7%, 103 ± 15%, 158 ± 20% and 168 ± 11% (Fig. 5); In Fig. 5, large yellow croaker feeding is containing the complement activity (ACH in Lactobacillus plantarum SaccharomycescerevisiaeP13 feed 1-4 week 50), the mean value that each numerical value equals every group of 3 measured values adds standard error, p ﹤ 0.05.
2.2 Lactobacillus plantarums improve the explosive breathing power of large yellow croaker leucocyte and phagocytic activity:
While attacking invasion pathogen by phagocyte when explosive breathing, produce, be usually used in evaluating the epidemic prevention ability of body to pathogen; Probio can improve the Haemocytes of Fishes such as rainbow trout outburst breathing power and head-kidney leukocytic phagocytic.Result of the present invention shows after the feed surrounding that feeding contains Lactobacillus plantarum (SaccharomycescerevisiaeP13), and the explosive breathing power of large yellow croaker head-kidney leucocyte and phagocytic activity are all apparently higher than the large yellow croaker (Fig. 6 and Fig. 7) of feeding control feed; In Fig. 6, large yellow croaker feeding is containing the head-kidney leucocyte outburst breathing power in Lactobacillus plantarum Saccharomyces cerevisiaeP13 feed 1-4 week.The mean value that each numerical value equals every group of 3 measured values adds standard error, p ﹤ 0.05.In Fig. 7, large yellow croaker feeding is containing the head-kidney leukocytes phagocytic activity in Lactobacillus plantarum SaccharomycescerevisiaeP13 feed 1-4 week.The mean value that each numerical value equals every group of 3 measured values adds standard error.p﹤0.05。
2.3 Lactobacillus plantarums improve large yellow croaker lysozyme activity:
The invasion that body is on the one hand resisted external foreign matter by immunocyte is also carried molecule and serial lysozyme strengthens the defence of body to pathogen by not mending simultaneously.In the present invention, feeding Lactobacillus plantarum 10 4, 10 6, 10 8with 10 10the large yellow croaker lysozyme activity of cfu/kg has had than control group and has significantly improved (Fig. 8), and in Fig. 8, large yellow croaker feeding is containing the serum lysozyme activities reached (ACH in Lactobacillus plantarum SaccharomycescerevisiaeP13 feed 1-4 week 50).The mean value that each numerical value equals every group of 3 measured values adds standard error.P﹤0.05。
The present invention is by containing Lactobacillus plantarum 0,1 × 10 to large yellow croaker feeding 4, 1 × 10 6, 1 × 10 8with 1 × 10 10after feed (table 1) surrounding of cfu/kg, measure respectively the immune parameter of reflection large yellow croaker innate immune activity.Comprehensive research result shows, Lactobacillus plantarum SaccharomycescerevisiaeP13 can be by improving renal cell outburst breathing power, phagocytic activity, complement vigor and the serum lysozyme activities reached of large yellow croaker, and the natural immune system of enhancing large yellow croaker, this provides strong empirical theory foundation with the research and development of biological feedstuff, the sound development of promotion large yellow croaker aquaculture for large yellow croaker cultivation.
The foregoing is only preferred embodiment of the present invention, not in order to limit the present invention, all any amendments of doing within the spirit and principles in the present invention, be equal to and replace and improvement etc., within all should being included in protection scope of the present invention.

Claims (5)

1. a large yellow croaker Lactobacillus plantarum feed formula, is characterized in that, this large yellow croaker Lactobacillus plantarum feed formula comprises: Lactobacillus plantarum 1 × 10 6the basal feed of the feed for nursing of cfu/kg, thalline 0.1g/kg, milk 9.999g/kg, fish meal 540g/kg, starch 183g/kg, fish cream 37g/kg, gluten 20g/kg, fish oil 20g/kg, shrimp shell meal 64g/kg, defatted soybean meal 94g/kg, mineral intermixture 35g/kg, vitamin mixtures 7g/kg composition and crude protein, thick fat, ash content and water composition;
According to mass percent, feed for nursing is 25.2%, crude protein 47.3%, thick fat 8.9%, ash content 10.2%, moisture 8.4%.
2. large yellow croaker Lactobacillus plantarum feed formula as claimed in claim 1, is characterized in that, this large yellow croaker Lactobacillus plantarum feed formula is: Lactobacillus plantarum 1 × 10 8cfu/kg, thalline 0.1g/kg, milk 9.99g/kg, fish meal 540g/kg, starch 183g/kg, fish cream 37g/kg, gluten 20g/kg, fish oil 20g/kg, shrimp shell meal 64g/kg, defatted soybean meal 94g/kg, mineral intermixture 35g/kg, vitamin mixtures 7g/kg, crude protein 47.3%, thick fat 8.9%, ash content 10.2%, moisture 8.4%.
3. large yellow croaker Lactobacillus plantarum feed formula as claimed in claim 1, is characterized in that, this large yellow croaker Lactobacillus plantarum feed formula is: Lactobacillus plantarum 1 × 10 10cfu/kg, thalline 10g/kg, fish meal 540g/kg, starch 183g/kg, fish cream 37g/kg, gluten 20g/kg, fish oil 20g/kg, shrimp shell meal 64g/kg, defatted soybean meal 94g/kg, mineral intermixture 35g/kg, vitamin mixtures 7g/kg, crude protein 47.3%, thick fat 8.9%, ash content 10.2%, moisture 8.4%.
4. a preparation method for large yellow croaker Lactobacillus plantarum feed, is characterized in that, the preparation method of this large yellow croaker Lactobacillus plantarum feed comprises the following steps:
Separate and obtain Lactobacillus plantarum from Korea S's pickled cabbage, in the sterile flask that 37 DEG C are incubated at containing MRS meat soup, after 24 hours, centrifugal 5 minutes of 15400 × g, collects Lactobacillus plantarum mixture;
Mix with skim milk in 1:4 ratio, freeze drying, by the viability of MRS agar plate numeration mensuration Lactobacillus plantarum mixture;
According to Lactobacillus plantarum 1 × 10 6cfu/kg, thalline 0.001g/kg, milk 9.999g/kg, fish meal 540g/kg, starch 183g/kg, fish cream 37g/kg, gluten 20g/kg, fish oil 20g/kg, shrimp shell meal 64g/kg, defatted soybean meal 94g/kg, mineral intermixture 35g/kg, vitamin mixtures 7g/kg configuration large yellow croaker feed for nursing;
Adding composition is the basal feed of 47.3% crude protein, 8.9% thick fat, 10.2% ash content and 8.4% moisture, is mixed and made into large yellow croaker Lactobacillus plantarum feed.
5. the preparation method of large yellow croaker Lactobacillus plantarum feed as claimed in claim 4, is characterized in that, this large yellow croaker Lactobacillus plantarum feed is fed large yellow croaker twice in one day.
CN201410188158.6A 2014-05-05 2014-05-05 Formula and preparation method of large yellow croaker lactobacillus plantarum feed Pending CN103947886A (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201410188158.6A CN103947886A (en) 2014-05-05 2014-05-05 Formula and preparation method of large yellow croaker lactobacillus plantarum feed

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201410188158.6A CN103947886A (en) 2014-05-05 2014-05-05 Formula and preparation method of large yellow croaker lactobacillus plantarum feed

Publications (1)

Publication Number Publication Date
CN103947886A true CN103947886A (en) 2014-07-30

Family

ID=51325357

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201410188158.6A Pending CN103947886A (en) 2014-05-05 2014-05-05 Formula and preparation method of large yellow croaker lactobacillus plantarum feed

Country Status (1)

Country Link
CN (1) CN103947886A (en)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN107874012A (en) * 2017-10-10 2018-04-06 国营无为县水产养殖场 A kind of breeding feed of large yellow croaker seedling

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH0239861A (en) * 1988-07-29 1990-02-08 Nippon Nousan Kogyo Kk Culture of sea urchins
CN1947548A (en) * 2005-10-11 2007-04-18 集美大学 Compound fish bait used specially for pseudosciaena crocea
CN102258130A (en) * 2010-05-28 2011-11-30 李荣帮 Preparation method of feed for large yellow croaker
CN103478449A (en) * 2013-08-22 2014-01-01 福建天马科技集团股份有限公司 Buoyancy puffing granulated compound feed for medium and adult pseudosciaena crocea
CN103583874A (en) * 2013-10-17 2014-02-19 蚌埠市丰牧牛羊肉制品有限责任公司 Compound feed for juvenile pseudosciaena crocea and preparation method thereof

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH0239861A (en) * 1988-07-29 1990-02-08 Nippon Nousan Kogyo Kk Culture of sea urchins
CN1947548A (en) * 2005-10-11 2007-04-18 集美大学 Compound fish bait used specially for pseudosciaena crocea
CN102258130A (en) * 2010-05-28 2011-11-30 李荣帮 Preparation method of feed for large yellow croaker
CN103478449A (en) * 2013-08-22 2014-01-01 福建天马科技集团股份有限公司 Buoyancy puffing granulated compound feed for medium and adult pseudosciaena crocea
CN103583874A (en) * 2013-10-17 2014-02-19 蚌埠市丰牧牛羊肉制品有限责任公司 Compound feed for juvenile pseudosciaena crocea and preparation method thereof

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
廖志勇，等: "植物乳杆菌Saccharomyces cerevisiae P13 对大黄鱼的促生长作用研究", 《安徽农业科学》 *
廖志勇，等: "植物乳杆菌对大黄鱼非特异性免疫力的影响", 《安徽农业科学》 *

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN107874012A (en) * 2017-10-10 2018-04-06 国营无为县水产养殖场 A kind of breeding feed of large yellow croaker seedling

Similar Documents

Publication Publication Date Title
Thompson et al. An in vitro and in vivo assessment of the potential of Vibrio spp. as probiotics for the Pacific White shrimp, Litopenaeus vannamei
Zhou et al. Effect of probiotic on larvae shrimp (Penaeus vannamei) based on water quality, survival rate and digestive enzyme activities
CN102041238B (en) Lactobacillus plantarum, method for fermenting and preparing bacteriocin of Lactobacillus plantarum, and application of Lactobacillus plantarum and bacteriocin
CN102399733B (en) Lactobacillus johnsonii, microbial inoculum, application and premix thereof
CN103555640B (en) Bacillus subtilis and application thereof to livestock breeding
CN105176874B (en) Bacillus coagulans FM603 and its application
CN105175518B (en) The bacteriocin and preparation method thereof that bacillus coagulans FM603 is generated
CN104106726A (en) New probiotic, and probiotic fermentation feed prepared by using it
JP2015530401A (en) Probiotic composition and prebiotic composition
CN103952324B (en) A kind of Kluyveromyces marxianus C2 and preparation method and application
CN103525718B (en) Bacillus cereus and probiotics powder thereof as well as preparation and application of probiotics powder
CN107858302A (en) A kind of bacillus subtilis 7K and its application
CN109874920A (en) A kind of composite microbial feed additive and preparation method thereof
Novriadi et al. Corn fermented protein in production diets for pacific white legged shrimp Litopenaeus vannamei: Improved growth performance, health status and resistance to infection
JP2013188196A (en) Virus infection prevention lactic acid bacterium composition and virus infection prevention lactic fermentation food
Duarte et al. Saccharomyces cerevisiae used as probiotic: Strains characterization and cell viability
CN105104712B (en) A kind of additive for microbe feedstuff and preparation method thereof
CN107937286A (en) A kind of Pichia anomala and its fermentation culture medium and application
CN103947886A (en) Formula and preparation method of large yellow croaker lactobacillus plantarum feed
CN103952357B (en) The saliva lactobacillus of a kind of efficient suppression fowl source Campylobacter
CN114085789B (en) Pediococcus pentosaceus MA.WTPQJ01 and application thereof
CN104212745A (en) Chicken microecological preparation and preparation method thereof
CN102994416A (en) Dog source bacillus licheniformis Y10 and application thereof
CN109385406A (en) One plant of vibrio parahaemolyticus phage and its application in terms of enhancing aquatic livestock immunity
CN106987543B (en) A kind of serratia marcescens BSFL-6 in stratiomyiid enteron aisle source and its application

Legal Events

Date Code Title Description
C06 Publication
PB01 Publication
C10 Entry into substantive examination
SE01 Entry into force of request for substantive examination
C02 Deemed withdrawal of patent application after publication (patent law 2001)
WD01 Invention patent application deemed withdrawn after publication

Application publication date: 20140730