CN108660086A - A kind of fish blood agar culture-medium and preparation method thereof - Google Patents
A kind of fish blood agar culture-medium and preparation method thereof Download PDFInfo
- Publication number
- CN108660086A CN108660086A CN201710194445.1A CN201710194445A CN108660086A CN 108660086 A CN108660086 A CN 108660086A CN 201710194445 A CN201710194445 A CN 201710194445A CN 108660086 A CN108660086 A CN 108660086A
- Authority
- CN
- China
- Prior art keywords
- culture
- agar
- culture medium
- fish
- fish blood
- 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
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- 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
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Genetics & Genomics (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Biotechnology (AREA)
- Wood Science & Technology (AREA)
- Microbiology (AREA)
- Medicinal Chemistry (AREA)
- Biomedical Technology (AREA)
- Virology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Tropical Medicine & Parasitology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The present invention provides a kind of fish blood agar culture-mediums, including peptone, beef extract powder, sodium chloride, de- fiber fish blood and agar, contain 5~20g of peptone, 1~10g of beef extract powder, 1~10g of yeast powder, 1~10g of sodium chloride, de- fiber 40~100ml of fish blood and agar 10~20g, pH ranging from 7.0 8.0 in wherein every 1000ml culture mediums.Culture medium cost prepared by the present invention is cheap, and preparation procedure is simple, while can stablize for the separation identification of paroxysmal aquatic products pathogenetic bacteria, has better applicability compared to existing sheep blood agar medium in the market.
Description
Technical field
The present invention relates to biotechnologies, and in particular to a kind of fish blood agar culture-medium formula, the formula are suitable for fish
The culture and screening of class pathogenic bacteria.
Background technology
Currently used pathogenic microorganism isolation and identification method is the typical illness fish of acquisition, through surface sterilization, sterile working
Take the lesion tissues sample such as liver, kidney, intestines, nutrient agar panel or blood agar plate scribing line, after 30 DEG C ± 2 DEG C are incubated overnight, picking
Suspicious bacterium colony pure culture.Blood agar plate is a kind of beef extract albumen containing de- fiber animal blood (general to use rabbit blood or sheep blood)
Peptone culture medium, therefore in addition to cultivating the required various nutrition of bacterium, moreover it is possible to coenzyme (such as V factors), ferroheme (the X factors) are provided
Etc. special growth factor.Blood agar culture-medium is usually used in detaching certain pathogenic microorganisms to nutritional requirement harshness, this in addition
Culture medium may further be used to measure the haemocylolysis of bacterium.
Currently used blood agar basal medium substantially (is soaked in nutrient agar by peptone, sodium chloride, beef
Powder, agar composition) on the basis of addition Sheep Blood be made.But this blood agar culture-medium shelf-life is very short, and cost is higher, purchase
Period is long, is not well positioned to meet separation, the qualification requirement of the paroxysmal disease of aquaculture.
Therefore, research cost is cheap, and preparation procedure is simple, while guaranteeing to stablize for the separation identification of aquatic products pathogenetic bacteria
Culture medium, be technical problem in the urgent need to address.
Invention content
The object of the present invention is to provide a kind of fish blood agar culture-medium, material is easy to get, and prepares simplicity, of low cost, makes it
It can be widely used in the separation identification of aquatic pathogenic bacterium, improve the determination rates of aquatic pathogenic bacterium.
The present invention provides a kind of fish blood agar culture-mediums, including peptone, beef extract powder, sodium chloride, de- fiber fish blood
And agar, wherein containing 5~20g of peptone, 1~10g of beef extract powder, 1~10g of yeast powder, chlorination in per 1000ml culture mediums
1~10g of sodium, de- fiber 40~100ml of fish blood and agar 10~20g, pH ranging from 7.0-8.0.
In the present invention, the peptone is to be made after pancreatin digests using beef and ox bone as raw material, and market has existing
Domestic or import commercially produced product.It is preferred that containing 10~15g of peptone in per 1000ml culture mediums.
In the present invention, for the beef extract powder using beef as raw material, beef original is made in thermally treated, filtering, concentration, drying
Powder, then by beef original powder by hydrolysis, be separated by solid-liquid separation, refrigeration, filtering, concentration and the processes such as dry obtain, be capable of providing organic
The nutriments such as acid, nucleotide, minerals, vitamin.There is existing domestic or import commercially produced product in market.It is preferred that every
Contain 3~8g of beef extract powder in 1000ml culture mediums.
In the present invention, the yeast extract using fresh yeast as raw material, be capable of providing B family vitamin and various amino acid with
And carbohydrate (predominantly glycogen and trehalose).There is existing domestic or import commercially produced product in market.It is preferred that being cultivated per 1000ml
Contain 3~8g of yeast powder in base.
In the present invention, sodium chloride is for maintaining osmotic pressure.It is preferred that containing 3~8g of sodium chloride in per 1000ml culture mediums.
In the present invention, the de- fiber fish blood in culture medium is the good nutrition substance of bacterial growth breeding, and fish blood can be selected from
Black carp, grass carp, silver carp, bighead, carp, crucian, murrel fish etc., preferably black carp, grass carp or crucian.The fish for choosing health takes blood, takes off
It is saved backup in 2-8 DEG C of refrigerator after fiber.50-80ml is added in preferably every 1000ml culture mediums when preparation and takes off fiber fish blood system
.
In the present invention, medium pH ranging from 7.0-8.0, more preferably 7.2-7.5.
Preferably, the present invention provides a kind of fish blood agar culture-mediums, wherein per in 1000ml culture mediums, contain albumen
Peptone 15g, beef extract powder 5g, yeast powder 5g, sodium chloride 5g, de- fiber fish blood 50ml and agar 15g, pH ranging from 7.2 ± 0.2.
The fish blood agar culture-medium formula of the present invention is capable of providing full nutrition, maintains bacterial penetration pressure.It is prepared by the present invention
Culture medium cost it is cheap, preparation procedure is simple, at the same can stablize for paroxysmal aquatic products pathogenetic bacteria separation identification and training
It supports, has better applicability compared to existing sheep blood agar medium in the market.
Specific implementation mode
Technical scheme of the present invention is further described with reference to embodiment, however, it is not limited to this, every right
Technical solution of the present invention is modified or replaced equivalently, and without departing from the spirit of the technical scheme of the invention and range, should all be contained
It covers in protection scope of the present invention.
Embodiment 1：It is prepared by culture medium
Aseptic aspiration healthy grass carp blood, is put into the vial for filling bead, notices that bead puts vial into advance
Mesohigh sterilizes and dries, and then shakes repeatedly about 10-20 minutes and wraps fibrin on bead until seeing, can
Stop, then dispensing in superclean bench, saved backup in 2-8 DEG C of refrigerator, can generally be preserved 2 weeks to one month.
If it find that haemolysis, indicates that the fish blood cannot be continuing with.
Blood agar culture-medium formula provided in this embodiment contains peptone, beef extract powder, yeast powder, sodium chloride, de- fibre
Tie up fish blood and agar.Wherein per 1000ml culture mediums in, containing peptone 15g, beef extract powder 5g, yeast powder 5g, sodium chloride 5g,
De- fiber fish blood 50ml and agar 15g, pH range 7.2 ± 0.2.
The preparation process of above-mentioned culture medium is as follows：Proportionally by peptone, beef extract powder, yeast powder, sodium chloride, agar
Mixing is added purified water constant volume and dissolves by heating and adjust pH value to 7.2 ± 0.2, sterilizes 15 minutes through 121 DEG C, be cooled to 50 DEG C,
It is proportionally added into sterile de- fiber fish blood, tablet is perfused after mixing, fish blood meida finished product is obtained after condensation, it is spare.
Embodiment 2：Pathogen is identified and isolated from
Illness grass carp mainly picks up from In Hangzhou Region of Zhe Jiang Province aquatic farm.The typical illness grass carp of acquisition, through surface sterilization, sterile behaviour
Take the lesion tissues sample such as liver, kidney, intestines, sheep blood agar tablet or the scribing line of fish blood plate, after 30 DEG C ± 2 DEG C are incubated overnight, picking
Suspicious bacterium colony pure culture.From culture experiment result, bacterium detection number is better than sheep blood agar tablet with state fish blood plate.From
One plant of tool typical case's β type haemolysis putative pathogen that fish blood plate culture experiment isolates and purifies, is denoted as SCSQ1415.
Gram's staining shows that SCSQ1415 is Gram-negative bacteria.Sterile working scrapes appropriate bacterium, fills GN+Card,
Upper machine identification.Physiological and biochemical analysis is the results show that SCSQ1415 is Aeromonas sobria.
Isolated strains SCSQ1415 mitochondria 16S rRNA amplified productions, electrophoresis verification, recycling, send raw work to be sequenced.Taking can
Become the relatively large number of forward primer sequencing result in site, phylogenetic analysis is the results show that SCSQ1415 and Wei Luona gas unit cells
Bacterium ATCC35624 16S rRNA affiliations are closer, are Aeromonas sobria (Aeromonas sobria), are under the jurisdiction of gas list
Born of the same parents Cordycepps (Aermonadaceae) Aeromonas (Aeromonas).The bacterium is widely present in water environment, is a variety of aquatic products
The main pathogenic bacteria of animal is also the bacterial pathogen of hemorrhagic disease of grass carp.
Claims (10)
1. a kind of fish blood agar culture-medium, including peptone, beef extract powder, yeast powder, sodium chloride, de- fiber fish blood and agar,
Wherein per 1000ml culture mediums in containing 5~20g of peptone, 1~10g of beef extract powder, 1~10g of yeast powder, sodium chloride 1~
10g, de- fiber 40~100ml of fish blood and agar 10~20g, pH ranging from 7.0-8.0.
2. culture medium according to claim 1, it is characterised in that：In the culture medium, contain in every 1000ml culture mediums
10~15g of peptone.
3. culture medium according to claim 1, it is characterised in that：In the culture medium, contain in every 1000ml culture mediums
3~8g of beef extract powder.
4. culture medium according to claim 1, it is characterised in that：In the culture medium, contain in every 1000ml culture mediums
3~8g of yeast powder.
5. culture medium according to claim 1, it is characterised in that：In the culture medium, contain in every 1000ml culture mediums
3~8g of sodium chloride.
6. culture medium according to claim 1, it is characterised in that：In the culture medium, takes off fiber fish blood and be selected from black carp, grass
Fish, silver carp, bighead, carp, crucian or murrel fish.
7. culture medium according to claim 6, it is characterised in that：In the culture medium, takes off fiber fish blood and be selected from black carp, grass
Fish or crucian.
8. culture medium according to claim 1, it is characterised in that：In the culture medium, it is added in every 1000ml culture mediums
50-80ml takes off fiber fish blood and is made.
9. culture medium according to claim 1, it is characterised in that：The medium pH ranging from 7.2-7.5.
10. a kind of fish blood agar culture-medium, wherein containing peptone 15g, beef extract powder 5g, yeast powder in per 1000ml culture mediums
5g, sodium chloride 5g, de- fiber fish blood 50ml and agar 15g, pH ranging from 7.2 ± 0.2.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710194445.1A CN108660086A (en) | 2017-03-29 | 2017-03-29 | A kind of fish blood agar culture-medium and preparation method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710194445.1A CN108660086A (en) | 2017-03-29 | 2017-03-29 | A kind of fish blood agar culture-medium and preparation method thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN108660086A true CN108660086A (en) | 2018-10-16 |
Family
ID=63786377
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710194445.1A Pending CN108660086A (en) | 2017-03-29 | 2017-03-29 | A kind of fish blood agar culture-medium and preparation method thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN108660086A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110819570A (en) * | 2019-11-27 | 2020-02-21 | 中秀科技股份有限公司 | Blood agar plate and preparation method thereof |
| CN113122423A (en) * | 2020-01-13 | 2021-07-16 | 李倩 | System of culture medium for pathogenic microorganism detection and preparation method thereof |
-
2017
- 2017-03-29 CN CN201710194445.1A patent/CN108660086A/en active Pending
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110819570A (en) * | 2019-11-27 | 2020-02-21 | 中秀科技股份有限公司 | Blood agar plate and preparation method thereof |
| CN113122423A (en) * | 2020-01-13 | 2021-07-16 | 李倩 | System of culture medium for pathogenic microorganism detection and preparation method thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Wegener et al. | Isolation of vancomycin resistant Enterococcus faecium from food | |
| CN105176874B (en) | Bacillus coagulans FM603 and its application | |
| CN103525718B (en) | Bacillus cereus and probiotics powder thereof as well as preparation and application of probiotics powder | |
| CN107338198A (en) | A kind of Lactobacillus plantarum and its application | |
| CN108660086A (en) | A kind of fish blood agar culture-medium and preparation method thereof | |
| CN105039202A (en) | Selective medium for multiplex proliferation of salmonella, listeria monocytogenes and vibrio parahaemolyticus and preparation method of selective medium | |
| CN114097927A (en) | Preparation method of biological feed additive based on bacteriophage | |
| CN101838626B (en) | Bartonia body solid-liquid double-phase slant culture medium as well as preparation method and application method thereof | |
| CN110205355A (en) | A kind of highly sensitive detection culture medium of microorganism and its preparation method and application | |
| JP5491683B2 (en) | Organic waste treatment using Bacillus microorganisms | |
| CN105104712B (en) | A kind of additive for microbe feedstuff and preparation method thereof | |
| CN111808771A (en) | Laboratory culture powder of American bacillus larvae of bees, preparation method and application thereof | |
| CN106701645A (en) | Bacillus amyloliquefaciens B7 with immune growth promoting effect and application method of bacillus amyloliquefaciens B7 | |
| Wijayanti et al. | DNA Barcoding of Swamp Sediment Bacterial Isolates for Swamp Aquaculture Probiotic | |
| CN109988811A (en) | A kind of microbial rapid detection culture medium and its preparation method and application | |
| CN110295211A (en) | A kind of preparation method and application of bacterium selenium-enriched protein | |
| CN105360873A (en) | Quick sterilization method of spirulina powder | |
| CN105639542A (en) | Chili base and preparation method thereof | |
| RU2518304C2 (en) | TWO-PHASE NUTRITIONAL MEDIUM FOR THIN LAYER CULTIVATION OF Helicobacter pylori AND METHOD OF ITS REALISATION | |
| CN102326779A (en) | Manufacturing method of drink being rich in bacillus natto and product thereof | |
| Prilipko et al. | Development of practical measures and ways of their realization for control, management of dairy raw materials and dairy products in accordance with eu norms | |
| Song et al. | Isolation, identification and thermal inactivation of dominant spoilage bacteria in egg curds | |
| US9809795B2 (en) | Extraction of nitrogen from organic materials through ammonification by mixed bacterial populations | |
| CN105838640A (en) | Composite probiotics for stichopus japonicus feeding and preparation method thereof | |
| CN110257299A (en) | A kind of application that enterobacter cloacae grows in promotion ruminant |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20181016 |
|
| WD01 | Invention patent application deemed withdrawn after publication |