CN110468069A - A kind of Lactobacillus casei YFI-5 and its application in anti-carp herpesviral II type - Google Patents
A kind of Lactobacillus casei YFI-5 and its application in anti-carp herpesviral II type Download PDFInfo
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- CN110468069A CN110468069A CN201910763441.XA CN201910763441A CN110468069A CN 110468069 A CN110468069 A CN 110468069A CN 201910763441 A CN201910763441 A CN 201910763441A CN 110468069 A CN110468069 A CN 110468069A
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- lactobacillus casei
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- carp
- crucian
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Classifications
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
- A23K10/16—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions
- A23K10/18—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions of live microorganisms
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K50/00—Feeding-stuffs specially adapted for particular animals
- A23K50/80—Feeding-stuffs specially adapted for particular animals for aquatic animals, e.g. fish, crustaceans or molluscs
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/66—Microorganisms or materials therefrom
- A61K35/74—Bacteria
- A61K35/741—Probiotics
- A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
- A61K35/747—Lactobacilli, e.g. L. acidophilus or L. brevis
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/12—Antivirals
- A61P31/20—Antivirals for DNA viruses
- A61P31/22—Antivirals for DNA viruses for herpes viruses
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
- C12N1/205—Bacterial isolates
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/11—Lactobacillus
- A23V2400/125—Casei
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/225—Lactobacillus
- C12R2001/245—Lactobacillus casei
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Abstract
The present invention relates to the antiviral probiotics technical fields of aquatic products, and in particular to a kind of Lactobacillus casei YFI-5 and its application in anti-carp herpes virus type 2.The deposit number of Lactobacillus casei of the present invention is the Lactobacillus casei YFI-5 fermented supernatant fluid in the CCTCC NO:M2019654 present invention, carp herpesviral II type (CyHV-2) can reduced to the infection activity of host cell, inhibit infection of the CyHV-2 poisoning intrusion to cell.To cytotoxic side effect.Feed addition Lactobacillus casei YFI-5 thallus can effectively reduce the death rate of crucian infection CyHV-2, can be used for preventing and treating crucian carp Hematopoietic Necrosis's disease.
Description
Technical field
The present invention relates to the antiviral probiotics technical fields of aquatic products, and in particular to a kind of Lactobacillus casei YFI-5 and
Its application in anti-carp herpes virus type 2.
Background technique
Crucian carp is one of China's cultured freshwater fish important species, and gross annual output amount is more than 3,000,000 tons, in freshwater aquiculture
In occupy highly important status.In recent years, China crucian leads to the bleeding for cultivating the death of hybridized prussian carp fulminant year after year
Property disease, causes heavy economic losses.Electronic Speculum observes the ultra-thin section of illness crucian spleen nephridial tissue, finds a large amount of typical blister sores
Malicious sample particle, further research confirms that the disease is crucian carp Hematopoietic Necrosis disease (Crucian Carp Hematopoietic
Necrosis), cause of disease is carp herpesviral II type (Cyprinid herpesvirus2, CyHV-2).Infect the different of CyHV-2
It educates pond crucian carp typical clinical symptom and is mainly shown as appetite stimulator, peel off only trip, and respiratory rate is accelerated, and vigor is poor, sinks to the bottom.Suffer from
Sick fish body table, fin base and gill bleeding, abdominal distention, exophthalmos.The gill filament is pale, and muscle has petechial hemorrhage, and spleen and kidney are rubescent swollen
Swollen, there is petechial hemorrhage, hepatomegaly, bleeding, enteron aisle sky in fish glue.Crucian carp Hematopoietic Necrosis's disease is a kind of highly infective disease, but its
Spread scope is limited, studies have shown that the disease is only to goldfish, crucian and its common mutation infectious at present.Researcher has found gold
The hybrid of fish and carp is also the susceptible body of CyHV-2, and the fish body of each specification can infect.
It is all puzzlement culture fishery development for a long time that the generation of disease how is prevented and treated in aquaculture process
A major issue.Currently, the method for preventing and treating aquiculture animal disease mainly has medical treatment and vaccine immunity two
Kind.However a series of problems, such as side effect caused by medical treatment, medicament residue, drug resistance and water pollution, is gradually by society
The most attention of meeting all circles, in addition to this, medical treatment is not fairly obvious for the therapeutic effect of viral disease；By sensitive thin
Born of the same parents be shortage, virus variation, immunization ways inconvenience influence, the development of aquaculture vaccines is affected.Therefore, it is necessary to constantly seek
The effective ways of controllable viral disease, probiotics because its is nontoxic, without drug resistance, noresidue, antibacterial, antiviral, growth promotion,
Green safe advantage receives significant attention.
In recent years, many researchers think that lactic acid bacteria is expected to a certain extent as drug in aquaculture process
Substitute is used for the prevention and treatment of various diseases, provides new strategy for disease prevention and control scheme.Yang Yongs etc. (2006) prove lactic acid
Bacterium metabolite has the inhibiting effect of highly significant to the growth of Vibrio anguillarum, inhibits efficiency 90% or more.Gildberg etc.
(1998) gadus seedling is fed with the feed of the lactic acid bacteria extracted containing cod fish guts, puts fry in a suitable place to breed after 3 months
In environment with strong morbid vibrio, disease-resistant rate can be improved.Existing data shows that lactic acid bacteria not only has antibacterial activity, also has
Antiviral activity.Ang etc. (2016) discovery Lactobacillus casei can be by inhibiting infecting for Coxsackie virus to prevent and treat brothers mouthful
Disease.Currently, lactic acid bacteria is widely used in aquaculture process, but aquatic pathogenic bacterium is inhibited to grind with viral about lactic acid bacteria
Study carefully also less.
The present invention filters out one plant of Lactobacillus casei for the first time can be inhibited to carp herpes virus type 2, for the disease
The prevention and treatment of poison provides new approaches.
Summary of the invention
The object of the present invention is to provide a kind of Lactobacillus casei YFI-5, the deposit numbers of the bacterial strain are as follows:
CCTCC NO:M2019654.
It is another object of the present invention to provide the applications of Lactobacillus casei YFI-5.
In order to achieve the above object, the present invention takes following technical measures:
Lactobacillus casei YFI-5 of the invention is isolated from the water sample of aquaculture region pond, specifically with life
Salt water is managed by after cultivation pond water sample dilution, is applied on BHI solid plate, is finally inverted plate, in constant incubator
In 30 DEG C culture for 24 hours.The colony inoculation for selecting different shape is isolated and purified on ordinary broth plate, and to different bacterium into
The measurement of row antiviral functions, be finally obtained one plant can anti-carp herpesviral II type bacterial strain, be named as YFI-5, bacterial strain YF
I-5 is accredited as Lactobacillus casei through physiological and biochemical property measurement and 16S rDNA sequence homology analysis.
Lactobacillus casei YFI-5 belongs to lactobacillus, and being suitble to growth temperature is 37 DEG C, through lactic acid bacteria culture medium (MRS)
White is formed after cultivating 48h, round, surface is smooth wet, the bacterium colony of neat in edge, protrusion；Can ferment fructose, galactolipin and Portugal
Grape sugar cannot utilize melibiose, raffinose and xylose, cannot decompose arginine and produce ammonia.
The bacterial strain was sent on August 19th, 2019 to China typical culture collection center preservation, classification naming: cheese cream
Bacillus (Lactobacillus casei) YFI-5, deposit number: CCTCC NO:M2019654, address: Wuhan, China Wuhan is big
It learns.
It is bad to be prepared into treatment or prevention crucian carp blood forming organ including the use of Lactobacillus casei for the application of Lactobacillus casei YFI-5
Incruable disease drug, or it is prepared into the drug for treating or preventing and infecting caused disease by carp herpesviral II type (CyHV-2), or
It is prepared into carp herpesviral II type antivirotic, or is prepared into aquatic animal feed additive.
Compared with prior art, the invention has the following advantages that
Anti- carp herpesviral II type (CyHV-2) is carried out to cell sense with probiotics lactobacillus casei YFI-5 in the present invention
Dye, Lactobacillus casei YFI-5 have compared with traditional antiviral chemicals as potential antiviral probiotics
Following advantage:
1, it has no toxic side effect, is noresidue, antibacterial, antiviral, growth promotion, green safe.
2, have many advantages, such as it is easy to use, safe, without immunological stress, high financial profit
3, enter fish body by way of oral, inhibit the invasion and proliferation of carp herpesviral II type (CyHV-2), effectively
Prevent and treat crucian carp Hematopoietic Necrosis disease.Lactobacillus casei YFI-5 fermentation supernatant in the present invention first plus after fermentation supernatant adds
CyHV-2 dilution adds 2 kinds of modes of fermentation liquid and CyHV-2 dilution to have direct repression, inhibiting rate difference to virus simultaneously
It is 34%, 27%.Lactobacillus casei YFI-5pH7.0 fermented supernatant fluid, to cytotoxic side effect.
4, feed addition Lactobacillus casei YFI-5 thallus can effectively reduce the death rate of crucian infection CyHV-2.
Detailed description of the invention
Fig. 1 is each group crucian respiratory burst power level schematic diagram.
Fig. 2 is each group crucian serum lysozyme level schematic diagram.
Fig. 3 is each group crucian Complement C_3 level schematic diagram.
Specific embodiment
Test method and condition are conventional method unless otherwise instructed in the embodiment of the present invention.These embodiments are only used
In illustrating that the present invention, protection scope of the present invention be not restricted by the embodiments.Technical solution of the present invention, such as not especially
Illustrate, is the conventional scheme of this field；The reagent or material derive from commercial channel if not otherwise specified.
Embodiment 1:
Bacterial strain YFI-5 separation identification
1, bacterial strain YFI-5 separation and identification
Lactobacillus casei is isolated from the water sample of aquaculture region pond, specifically with 0.85% sterile physiological
Salt water will cultivate pond water sample continuous 10 times of doubling dilutions 6 times, be inhaled in each concentration gradient dilution respectively with liquid-transfering gun
It takes on 100 μ L to BHI solid plate of solution, is smoothened, numbered with spreading rod, and do 3 repetitions.In ultra-clean work after coating uniformly
5~10min is placed in platform, is fully absorbed media surface bacterium solution.Finally plate is inverted, 30 DEG C in constant incubator
Culture is for 24 hours.The colony inoculation for selecting different shape is isolated and purified on ordinary broth plate, and is carried out to different bacterium disease-resistant
The measurement of malicious function, be finally obtained one plant can anti-carp herpesviral II type bacterial strain, be named as YFI-5.
2, the identification of YFI-5 bacterial strain
1), physiological and biochemical property
Learn from else's experience the bacterial strain YFI-5 of MRS solid medium pure culture, and single colonie streak inoculation is on BUG identification plate, and 30 DEG C
16h-24h is cultivated, Biolog bacterial identification kits IF-A inoculation liquid is taken when bacterium colony suitable size, pipe outer wall is wiped
Only, being placed in Biolog transmissometer and adjusting its reading is 100%T；Suitable single bacterium is dipped with aseptic cotton carrier to drop down onto inoculation liquid,
Make transmissometer reading between 92%T-98%T, mixed liquor is transferred to GEN III with every 100 μ L volume of hole with 8 hole liquid-transfering guns and is reflected
In fixed board, then identification plate is loaded into Biolog system and is cultivated, system automatic reading simultaneously exports qualification result.Physiology and biochemistry
Feature is shown in Table 1.
The physiological and biochemical property of 1 bacterial strain YFI-5 of table
2) molecular biology identification of bacterial strain YFI-5
The gene for expanding bacterial strain 16SrRNA uses universal primer, 16SF (27F): AGAGTTTGATCMTGGCTCAG, 16SR
(1492R): GGTTACCTTGTTACGACTT is synthesized by Shanghai Sangon Biological Engineering Technology And Service Co., Ltd.Bacterial strain YFI-5
Through physiological and biochemical property measurement and 16S rDNA sequence homology analysis, it is accredited as Lactobacillus casei, the bacterial strain is in 2019 8
The moon is sent to China typical culture collection center preservation, classification naming on the 19th: Lactobacillus casei (Lactobacillus casei)
YFI-5, deposit number: CCTCC NO:M2019654, address: Wuhan, China Wuhan University.
Embodiment 2: Lactobacillus casei YFI-5 fermented supernatant fluid antiviral spectrum detection
Mono- single colonie of Lactobacillus casei YFI-5 of picking plate rejuvenation is inoculated in 150ml MRS fluid nutrient medium, and 37
DEG C culture for 24 hours, 5000rpm be centrifuged 10min, take 0.22 μm of membrane filtration of supernatant, it is standby to be stored in 4 DEG C of preservations in sterile centrifugation tube
With.Detection Lactobacillus casei YFI-5 fermented supernatant fluid is to carp herpesviral II type (CyHV-2), giant salamander irido virus respectively
(GSIV), grass carp reovirus (GCRV), the inhibitory effect of huichun viremia virus (SVCV).
Cell used in this implementation the present embodiment is crucian carp brain tissue cell system (GiCB, CCTCC NO:C2013179), giant salamander
Muscle cell system (GSM), grass carp kidney cell system (CIK), carp epithelial cell line (EPC).
The Lactobacillus casei YFI-5 fermented supernatant fluid volume being added in the present embodiment is every 100 μ L of hole, addition
100TCID50The virus liquid of/0.1ml and the volume of fermented supernatant fluid are identical.
Lactobacillus casei YFI-5 fermented supernatant fluid and CyHV-2 are accessed simultaneously in 96 well culture plates for growing up to single layer respectively
GiCB cell, the GSM that Lactobacillus casei YFI-5 fermented supernatant fluid and GSIV access grow up in 96 well culture plates of single layer is thin
Born of the same parents, Lactobacillus casei YFI-5 fermented supernatant fluid and GCRV access grow up to the CIK cell in 96 well culture plates of single layer, buttermilk bar
Bacterium YFI-5 fermented supernatant fluid and SVCV access grow up to the EPC cell in 96 well culture plates of single layer, and control group only accesses equivalent disease
Poison co-cultures 90min, abandons mixed liquor and is washed with PBS, and cell maintenance medium is added and continues to cultivate.It is seen under optics inverted microscope
Cytopathy (CPE) is examined, the antiviral effect of Lactobacillus casei YFI-5 fermented supernatant fluid is judged according to CPE quantity, the results are shown in Table
2 ,+indicate resistant ,-indicate non-resistant.
The antiviral spectrum of 2 Lactobacillus casei YFI-5 of table
Embodiment 3:
Application of the Lactobacillus casei YFI-5 fermented supernatant fluid in anti-carp herpesviral II type (CyHV-2):
Mono- single colonie of Lactobacillus casei YFI-5 of picking plate rejuvenation is inoculated in 150ml MRS fluid nutrient medium, and 37
DEG C culture for 24 hours, 5000rpm be centrifuged 10min, take 0.22 μm of membrane filtration of supernatant, it is standby to be stored in 4 DEG C of preservations in sterile centrifugation tube
With for the present embodiment and embodiment 4.
Cell used in the present embodiment is crucian carp brain tissue cell system (GiCB), CCTCC NO:C2013179.Cell used
Maintaining liquid is the M199 culture medium of 10%FBS；
The Lactobacillus casei YFI-5 fermented supernatant fluid volume being added in the present embodiment is every 100 μ L of hole, addition
100TCID50The CyHV-2 virus liquid of/0.1ml is identical as the volume of fermented supernatant fluid；
Virus titer detection: it is 1 × 10 that density is added into 96 porocyte culture plates4The GiCB cell 100 μ L in/hole, 25
DEG C culture 16~for 24 hours.When cell grows to 80%~90%, being inoculated with dilution thereto is 101~1010100 μ of virus liquid
The hole L/, each dilution are arranged 8 parallel holes, cultivate 2h in 25 DEG C of incubators.After incubation, virus liquid is recycled, is used
M199 culture medium rinses hole inner cell 2 times, and 100 μ L cell maintenance mediums of addition continue to cultivate 96h.3 groups of experimental setup are parallel, every
The CPE phenomenon of each dilution cell monolayer is observed and recorded for 24 hours, and records corresponding lesion hole count, is calculated according to Reed-Muench method
The median tissue cell infection amount (tissue culture infective dose, TCID50) of CyHV-2.
Experimental group is as follows:
1 group: using Lactobacillus casei YFI-5 fermented supernatant fluid pretreated group, then access virus: Lactobacillus casei YFI-5 hair
Ferment supernatant is inoculated in the cell for growing up to 96 well culture plates of single layer and co-cultures 2h, with 100TCID after washing50/ 0.1ml's
CyHV-2 infects cell monolayer, sets and adsorbs 90min in incubator, cell maintenance medium is added after washing, continues to cultivate.
2 groups: Lactobacillus casei YFI-5 fermented supernatant fluid and CyHV-2 access cell simultaneously: with 100TCID50/0.1ml
CyHV-2 is added after mixing in equal volume to be grown up in the cell of 96 well culture plates of single layer, and 90min is co-cultured, and is abandoned mixed liquor and is used in combination
PBS washing is added cell maintenance medium and continues to cultivate.
3 groups: first plus virus infected cell, then accessing Lactobacillus casei fermented supernatant fluid: with 100TCID50/0.1ml
CyHV-2 infection grows up to the cell of 96 well culture plates of single layer, washs after adsorbing 90min in incubator, adds Lactobacillus casei
Fermented supernatant fluid, PBS is washed after setting incubator culture 90min, and cell maintenance medium is added and continues to cultivate.
4 groups: the cell that Lactobacillus casei supernatant and 96 well culture plates for growing up to single layer is added co-cultures 2h, and rear addition is thin
Born of the same parents' maintaining liquid continues to cultivate.
5 groups: 100TCID50/ 0.1ml CyHV-2 infection grows up to the cell of 96 well culture plates of single layer, sets in incubator and adsorbs
PBS is washed after 90min, and cell maintenance medium is added and continues to cultivate
6 groups: normal cell controls.
By mtt assay indirect determination Lactobacillus casei YFI-5 fermented supernatant fluid to the inhibiting rate of CyHV-2 after 48h,
Viral suppression=(lactic acid bacteria processing group OD490Virus control group OD490)/(cell controls group OD490Virus
Control group OD490) × 100%
Specific inhibiting rate is as shown in table 3:
3 Lactobacillus casei YFI-5 fermented supernatant fluid of table is to CyHV-2 inhibiting rate
Embodiment 4:
Application of the Lactobacillus casei YFI-5 in preparation carp herpesviral II preparation:
1) crucian (20 ± 2g) is temporarily supported 2 weeks in laboratory conditions before experiment, daily 8 points and 18 points feed, feeding volume
It is the 1% of fish body weight, and continues to be oxygenated；Experimental water is to be aerated tap water, 25 ± 1 DEG C of water temperature, dissolved oxygen > 5mgL-1.pH it is
7.3 ± 0.5 (Cheng Jie 2011)；Crucian is tested through detecting virus-free and bacterium infection.
8 groups are divided into, every group of 60 tails.Following processing is done respectively:
1 group: 100 μ l CyHV-2 (10 are injected intraperitoneally in crucian7TCID50), 100 μ l Lactobacillus caseis are injected intraperitoneally after 48h
YFI-5 fermented supernatant fluid；
2 groups: 100 μ l Lactobacillus casei YFI-5 fermented supernatant fluids are injected intraperitoneally in crucian, and 100 μ l are injected intraperitoneally after 48h
CyHV-2(107TCID50)；
3 groups: injecting 100 μ l CyHV-2 (10 simultaneously in crucian abdominal cavity7TCID50) and 100 μ l Lactobacillus casei YFI-5 fermentation
The mixed liquor of supernatant；
4 groups: crucian feeds feed 2 days (10 of the YFI-5 containing Lactobacillus casei7Cfu/g) feeding volume is the 1% of fish body weight,
Then 100 μ l CyHV-2 (10 are injected intraperitoneally7TCID50)；
5 groups: 100 μ l CyHV-2 (10 are injected intraperitoneally in crucian7TCID50), start after 48h to feed YFI-5 containing Lactobacillus casei
Feed (107Cfu/g) feeding volume is the 1% of fish body weight.
6 groups: 100 μ l CyHV-2 (10 are injected intraperitoneally in crucian7TCID50), start simultaneously at feeding YFI-5 containing Lactobacillus casei
Feed (107Cfu/g) feeding volume is the 1% of fish body weight.
7 groups: 100 μ l CyHV-2 (10 are injected intraperitoneally in crucian7TCID50)；
8 groups: the processing of 100 μ l Lactobacillus casei YFI-5 fermented supernatant fluids is injected intraperitoneally in crucian.
9 groups: the 100 sterile PBS of μ l are injected intraperitoneally in crucian.
Each group crucian records in 14 days death condition (table 4) in death condition from packet transaction day.And on 0th and
It detects within 14th
Each group crucian related immune index: respiratory burst vigor (Fig. 1), serum lysozyme activities reached (Fig. 2), serum complement C3
Horizontal (Fig. 3).
Protective rate calculation formula:
Protective rate=(V'-V)/V' × 100%
V': the death rate (7 groups) after crucian direct injection CyHV-2；
V: the lactic acid bacteria processing group death rate.
Respiratory burst vigor testing methods:
The measurement of respiratory burst power is carried out according to the method for Anderson description: NBT reduction method (Anderson,
Brubacher et al.1998)。
The measurement of serum lysozyme activities reached
(Ellis 1988) that the measurement of serum lysozyme activities reached is measured with reference to the turbidity method of Ellis description.One
A antalzyme activity unit definition are as follows: 1min can make the amount of the lysozyme of absorptance decline 0.001 under the wavelength of 530nm.
The measurement of serum complement C3
The measurement of serum complement C3 level uses Nanjing to build up Complement C_3 assay kit, and (bio-engineering research is built up in Nanjing
Institute).As a result it is indicated as unit of mg/mL.
4 Lactobacillus casei YFI-5 of table is to crucian protective rate
Claims (5)
1. a kind of isolated Lactobacillus casei, the deposit number of the Lactobacillus casei is CCTCC NO:M2019654.
2. Lactobacillus casei described in claim 1 treats or prevents the application in crucian carp Hematopoietic Necrosis disease drug in preparation.
It is infected 3. Lactobacillus casei described in claim 1 is treated or prevented in preparation by carp herpesviral II type (CyHV-2)
Application in the drug of caused disease.
4. application of the Lactobacillus casei described in claim 1 in preparation carp herpesviral II type antivirotic.
5. Lactobacillus casei described in claim 1 is preparing the application in aquatic animal feed additive.
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