CN106480161A - A kind of mastitis for milk cows detection kit - Google Patents
A kind of mastitis for milk cows detection kit Download PDFInfo
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- CN106480161A CN106480161A CN201611009459.3A CN201611009459A CN106480161A CN 106480161 A CN106480161 A CN 106480161A CN 201611009459 A CN201611009459 A CN 201611009459A CN 106480161 A CN106480161 A CN 106480161A
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
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Abstract
The present invention relates to detection technique field, more particularly to a kind of mastitis for milk cows detection kit.The mastitis for milk cows detection kit includes：The common chromogenic culture medium of Gram-negative bacteria chromogenic culture medium, gram-positive bacteria chromogenic culture medium and enterococcus gold-coloured staphylococci；Gram-negative bacteria chromogenic culture medium includes first foundation culture medium and the first powdered beef base；Gram-positive bacteria chromogenic culture medium includes the second basal medium, the second powdered beef and sodium chloride；Chromogenic culture medium includes enterococcus basal medium and staphylococcus aureus basal medium to enterococcus gold-coloured staphylococci altogether.The mastitis for milk cows detection kit that the present invention is provided can quick and precisely detect recessive type mastitis for milk cows, it is not necessary to by specific apparatus, visually just can direct judged result, technical staff is easy to grasp, easy to operate, can go out result in 18h～24h, while can be with batch detection, low cost.
Description
Technical field
The present invention relates to detection technique field, more particularly to a kind of mastitis for milk cows detection kit.
Background technology
Mastitis for milk cows (Mastitis) is that cow mammary gland is subject to the stimulation of the factor such as physics, chemistry, microorganism caused
The inflammation of cow breast, shows as change, breast tissue generation pathology change that milk occurs physicochemical property and cytology property
Change, be one of regular incidence of milk cow.Clinical visible change is had or not according to breast and milk mammitis is divided into clinical type breast
Scorching (Clinical Mastitis) and subclinical mastitis (Subclinical Mastitis).Recessive mastitis shows as suffering from
The breast of ox and milk visually observe without exception, need by Somatic Cell Count or by other diagnostic methods just detectable breast
Room inflammation.
Count according to international milk cow federation, 20 century 70s, milk cow clinic mastitis illness rate about 2%, recessive breast
Room inflammation infected cattle is up to 50%；In 11,000,000 cow heads that the U.S. raises, half suffers from all kinds mammitis；The Japan whole nation is average
Mammitis illness rate is 45.1%.The incidence of China's mammitis is higher, typically in 20%-70%.The Chinese Academy of Agricultural Sciences Chinese veterinarian
Research institute carries out clinical type breast to northwest, North China, northeast, the adult cow in milk in 32, the city in South China 22 cattle farm 1037
The room inflammation cause of disease and incidence investigation, and wherein 5 20, areas cattle farm 3384 cow in milk is tested with Lanzhou mammitis
Recessive mastitis investigation is carried out, as a result shows that the clinic mastitis incidence of disease is 33.41%, recessive mastitis milk ox head positive rate
For 73.91%, positive udder region recall rate is 44.74%.It is reported that, the recessive mastitis positive rate of Beijing and Efficiency in Buildings in Tianjin Area is
50%-80.9%；The incidence of disease of the recessive mastitis in Lanzhou and Taiyuan is 50% or so.Qinghai Province cattle farm recessive mastitis
Nipple illness rate and newborn area's illness rate be respectively 54.08% and 25.89%.In sum, mastitis for milk cows is in world wide
Interior infection rate is all very high, and also quite serious in the infection conditions of China's mastitis for milk cows, mammitis infection type is mainly with recessiveness
Based on type.
Mammitis brings huge harm to milk cow production, the nutritive value of milk and food security.Mammitis first
Cause serious economic loss, the conventional estimated of the economic loss that foreign countries cause to mastitis for milk cows is the drop according to milk yield
The increase of the loss of low, milk, medication expense, animal doctor's expense and labour's expense etc. is counting.It is reported that, Britain treats milk every year
The average cost of garget is 187 pounds of every cow head, and it is 177-182 that the U.S. is used for the cost per cow head mammitis every year
Dollar, Finland, Norway, Sweden are every year because the milk cow that mammitis problem is eliminated accounts for 35%, 19% and the 22% of Culled cow, it is seen that
The economic loss that the generation of mammitis is brought to milk cow production is huge.In these economic losses, due to the reduction of the output of milk
The loss for causing accounts for 70%, so that milk cow is eliminated ahead of time and accounts for 14%, and discarded milk accounts for 7%, treats and animal doctor's expense
8% etc. is accounted for, and economic loss mostlys come from recessive mastitis, its loss for causing accounts for the annual total productive capacity of cow
10-11%.Secondly, mammitis is greatly reduced the quality of breast, contains rich in protein, fat and sugar class, in breast in breast
During scorching disease, pathogenic bacteria damage the secretory cell of mammary gland, cause breast tissue to damage, and some nutrient contents are lost, and make nutrition
The health-care effect that composition is not complete and reduces milk.In recessive mastitis milk, total amino acid content is also by average 2.2387g/100mL
Reduce 21%, wherein methionine, isoleucine, leucine, phenylalanine, 5 kinds of essential amino acids of lysine and histidine, essence
Propylhomoserin is significantly reduced；Lactose, casein, fat, Ca2+、P3-、K+Content Deng beneficiating ingredient all reduces or substantially reduces；And exempt from
The content of the harmful components such as the toxin of epidemic disease globulin, esterase, inflammatory factor, pathogenic bacteria and its generation then increases, and heat endurance drops
Low.Additionally, milk cow is with the poison containing substantial amounts of inflammatory factor, pathogenic bacteria and its generation in the milk produced after mammitis disease
Element, if drunk and sterilizing tight or having mixed up the milk containing pathogenic bacteria, can make one to feel not quite the thing, and severe patient can be lured
Send out disease.Treatment mammitis lysis in there is abuse of antibiotics, some operators eliminate treatment during suffer from cow's milk
Juice, causes antibiotic in Residues in Milk, causes eater that allergic reaction occurs, especially can more to the elderly and infant's harm
Greatly.Therefore, in time, make diagnosis exactly to mastitis for milk cows disease particularly important.
Clinical type mastitis for milk cows is mainly diagnosed by clinical symptoms, and recessive mastitis is then mainly examined by auxiliary
Disconnected method is monitored.The incidence of disease of mammitis is very high, especially recessive mastitis, and vaccary will be monitored at any time, make in time
Diagnosis, so as to cluster prevention.According to milk cow suffer from mammitis have a feature of cytosis in milk and establish direct Detection Method and
Indirect detection method.Direct Detection Method mainly has body cell direct counting method, electronic counting method, fluorescent electronic cell counting etc.,
Be most frequently with present at home for body cell direct counting method.Pathological research confirms, after breast infection inflammation, a large amount of white thin
Born of the same parents' entrance mammary gland, part epithelial cell shedding, so that somatic number in milk is significantly raised.According to this principle, per milliliter is calculated
Somatic number in milk, this method testing result are the most accurate, and this is the benchmark for diagnosing recessive mastitis, and other diagnosis sides
The standard that method is compared.Indirect method is current clinically most common method, and which is divided into chemical measure and physical measure, its
In at present clinically the most commonly used with chemical measure.Chemical measure is indirect determination latex cell number and milk pH value
Method.When room inflammation occurs, the pH value of milk rises, and just can reach the purpose of judgement by determining milk pH value.But these inspections
Survey method is required to special detecting instrument, and complex operation is had high demands to operating personnel.Accordingly, it is desirable to provide a kind of operation letter
List, fast and accurately mastitis for milk cows detection method.
Content of the invention
In view of this, the invention provides a kind of mastitis for milk cows detection kit.The mastitis for milk cows detection kit
Mastitis for milk cows can quick and precisely be detected, it is not necessary to by specific apparatus, visually just can direct judged result, technical staff is very
It is easily mastered, while can be with batch detection.
In order to realize foregoing invention purpose, the present invention provides technical scheme below：
The invention provides a kind of mastitis for milk cows detection kit, including：Gram-negative bacteria chromogenic culture medium, leather are blue
Family name's positive bacteria chromogenic culture medium and the common chromogenic culture medium of enterococcus gold-coloured staphylococci；
Gram-negative bacteria chromogenic culture medium includes first foundation culture medium and the first powdered beef base；First foundation culture medium
Gram-negative bacterium culture medium is praised for French Kerma (unit of kinetic energy)；
Gram-positive bacteria chromogenic culture medium includes the second basal medium, the second powdered beef and sodium chloride；Second basis
Culture medium praises Gram-positive bacterium culture medium for French Kerma (unit of kinetic energy)；
Chromogenic culture medium includes enterococcus basal medium and staphylococcus aureus base to enterococcus gold-coloured staphylococci altogether
Basal culture medium.
Preferably, in Gram-negative bacteria chromogenic culture medium, the concentration of first foundation culture medium is 10～20g/L, the
The concentration of one powdered beef is 2～4g/L；
In gram-positive bacteria chromogenic culture medium, the concentration of the second basal medium is 10～20g/L, the second powdered beef
Concentration is 1～3g/L, and the concentration of sodium chloride is 1～2g/L.
Preferably, in Gram-negative bacteria chromogenic culture medium, the concentration of first foundation culture medium is 15g/L, the first beef
The concentration of powder is 3g/L；
In gram-positive bacteria chromogenic culture medium, the concentration of the second basal medium is 15g/L, the concentration of the second powdered beef
For 2g/L, the concentration of sodium chloride is 1.5g/L.
Preferably, the pH value of Gram-negative bacteria chromogenic culture medium is 7.0 ± 0.2.
Preferably, the pH value of gram-positive bacteria chromogenic culture medium is 6.9 ± 0.2.
Preferably, the pH value of the common chromogenic culture medium of enterococcus gold-coloured staphylococci is 7.0 ± 0.2.
In the embodiment that the present invention is provided, in parts by weight, in first foundation culture medium：15 parts of agar, peptone 7
Part, 10 parts of dusty yeast, 1.2 parts of pigment.
In the embodiment that the present invention is provided, in parts by weight, in the second basal medium：15 parts of agar, peptone 8
Part, 12 parts of dusty yeast, 5 parts of salt, 4.4 parts of pigment, 2.0 parts of enriching substance.
In the embodiment that the present invention is provided, in parts by weight, enterococcus basal medium includes：Nutriment 40.1
Part, 1 part of Tween 80,0.25 part of developer, 15 parts of agar.The culture medium buys biology enterococcus colour developing culture rich in Qingdao sea
Base.
In the embodiment that the present invention is provided, in parts by weight, staphylococcus aureus basal medium includes：Nutrients
25 parts of matter, 1 part of Tween 80,40.3 parts of substance that show color, 15 parts of agar, 6 parts of antipathogenic composition.The culture medium is bought in Qingdao Hai Bo
Biological enterococcus chromogenic culture medium.
Preferably, the mass ratio of enterococcus basal medium and staphylococcus aureus basal medium for (10～
15)：(1～5).
Preferably, enterococcus basal medium is (11～12) with the mass ratio of staphylococcus aureus basal medium：
(4～5).
Preferably, mastitis for milk cows detection kit also includes vertically dividing equally check into three culture dish.
The invention provides a kind of mastitis for milk cows detection kit.The mastitis for milk cows detection kit includes：Leather is blue
The common chromogenic culture medium of family name's negative bacterium chromogenic culture medium, gram-positive bacteria chromogenic culture medium and enterococcus gold-coloured staphylococci；Leather
Lan Shi negative bacterium chromogenic culture medium includes first foundation culture medium and the first powdered beef base；Gram-positive bacteria chromogenic culture medium bag
Include the second basal medium, the second powdered beef and sodium chloride；Chromogenic culture medium includes enterococcus to enterococcus gold-coloured staphylococci altogether
Basal medium and staphylococcus aureus basal medium.The present invention has the advantages that：
1st, the mastitis for milk cows detection kit that the present invention is provided can quick and precisely detect recessive type mastitis for milk cows, be not required to
Will by specific apparatus, visually just can direct judged result, technical staff is easy to grasp, easy to operate, in 18h～24h
Result can be gone out, while can be with batch detection, low cost；
2nd, in the mastitis for milk cows detection kit that the present invention is provided, Gram-negative bacteria chromogenic culture medium and gram are positive
Property bacterium chromogenic culture medium can be remarkably promoted the growth of bacterium, can shorten detection time after improvement；
3rd, enterococcus gold-coloured staphylococci is altogether in chromogenic culture medium, enterococcus basal medium and staphylococcus aureus base
After basal culture medium is combined in specific proportions, enterococcus and staphylococcus aureus can be detected simultaneously.
Description of the drawings
Fig. 1 show the present invention prepare including mastitis Gram-negative bacteria chromogenic culture medium, mastitis gram-positive bacteria
Chromogenic culture medium and the vertically dividing equally check into three culture dish of the common chromogenic culture medium of enterococcus Staphylococcus aureus；
Fig. 2 shows the Pathogen detection result using culture medium of the present invention to mastitis for milk cows；
Fig. 3 shows the testing result of the emulsion using culture medium of the present invention detection flying crane emulsion pasture collection；
Fig. 4 shows the comparison of Gram-negative bacteria chromogenic culture medium of the present invention and the Detection results before improvement；
Fig. 5 shows the comparison of gram-positive bacteria chromogenic culture medium of the present invention and the Detection results before improvement；
Fig. 6 shows the Detection results of enterococcus of the present invention and the common chromogenic culture medium of gold-coloured staphylococci.
Specific embodiment
The invention discloses a kind of mastitis for milk cows detection kit, those skilled in the art can use for reference present disclosure,
It is suitably modified technological parameter realization.Specifically, all similar replacements and change come to those skilled in the art
Say be it will be apparent that they are considered as including in the present invention.The method of the present invention and application have passed through preferred embodiment
Be described, related personnel substantially can in without departing from present invention, spirit and scope to method described herein and should
Be modified or suitably change and combine, realize and apply the technology of the present invention.
Term is explained：
1st, mastitis for milk cows
Cow breast is subject to mechanical irritation, pathogenic microorganism immersion and chemicals rationality to damage and drawn during galactopoiesis
The breast pathological change for rising, is divided into serosity mammitis, cellulosic catarrh mammitis, suppurative mammitis, hemorrhagic breast
Scorching, gangrenous mastitis and recessive mastitis.
2nd, pathogenic infection
The infection caused by parasite or microorganism, so far, people have isolated 150 from cow mammary gland tissue
Kind of pathogenic microorganism, modal have 23 kinds, wherein 14 kinds of bacterium, 2 kinds of mycoplasma, fungi and 7 kinds of virus.Wherein the incidence of disease is most
High is staphylococcus aureus, Escherichia coli, streptococcus, and mycoplasma, the fungus-caused mammitis incidence of disease be year by year in recent years
Rise.
3rd, microbiological culture media
Nutrient matrix for microbial growth.It is divided into fluid nutrient medium and solid medium.Typically, in Liquid Culture
Base adds agar Solid agar culture is obtained.Culture medium has different formulas typically all to contain carbon source, nitrogen source and inorganic salts.
In the mastitis for milk cows detection kit that the present invention is provided, used medium raw material all can be buied by market.
With reference to embodiment, the present invention is expanded on further：
Embodiment 1
1. mastitis Gram-negative bacteria chromogenic culture medium is prepared
The present invention improvement after Gram-negative bacteria chromogenic culture medium formula be：15g/L basal medium+3g/L beef
Powder.
Wherein, basal medium：Agar 15.0g/L, peptone 7g/L, dusty yeast 10g/L, pigment 1.2g/L, pH value 7.0
± 0.2, buy Gram-negative bacterium culture medium is praised in French Kerma (unit of kinetic energy).
Preparation method is as follows：
(1) basal medium and powdered beef are weighed by above-mentioned formula, is dissolved in the clean triangular flask of 1000mL deionized water.
Also dependent on needing proportionally to expand or shrink the amount for preparing culture medium.
(2) 100 DEG C are heated to, do not stop stirring so as to be completely dissolved.It is sure not to be heated beyond 100 DEG C.If being added using micro-wave oven
Culture medium ebuillition of heated should be removed, gently shake up by heat immediately, place into microwave-oven-heating, and observation minute bubbles are changed into air
Bubble, until be completely dissolved.It is sure not to overflow culture medium.
(3) 45 DEG C～50 DEG C are cooled to the culture medium for having dissolved, are gently mixed, pour plate so as to solidify, dry standby
With.
2. mastitis gram-positive bacteria chromogenic culture medium is prepared
The present invention improvement after gram-positive bacteria chromogenic culture medium formula be：15g/L basal medium+2g/L powdered beef
+ 1.5g/L sodium chloride.
Wherein, basal medium：Agar 15.0g/L, peptone 8g/L and dusty yeast 12g/L, salt 5.0g/L, pigment
4.4g/L；Enriching substance：2.0g/L；PH value 6.9 ± 0.2, buys and praises Gram-positive bacterium culture medium in French Kerma (unit of kinetic energy).
Preparation method is as follows：
(1) basal medium, powdered beef and sodium chloride are weighed by above-mentioned formula, is dissolved in the cleaning three of 1000mL deionized water
In the bottle of angle, it is sufficiently stirred for mixing.Also dependent on needing proportionally to expand or shrink the amount for preparing culture medium.
(2) 100 DEG C are heated to, do not stop stirring so as to be completely dissolved.It is sure not to be heated beyond 100 DEG C.If being added using micro-wave oven
Culture medium ebuillition of heated should be removed, gently shake up by heat immediately, place into microwave-oven-heating, and observation minute bubbles are changed into air
Bubble, until be completely dissolved.It is sure not to overflow culture medium.
(3) 45 DEG C～50 DEG C are cooled to the culture medium for having dissolved, are gently mixed, pour plate so as to solidify, dry standby
With.
3. chromogenic culture medium is prepared altogether for enterococcus and gold-coloured staphylococci
(1) enterococcus basal medium：Nutriment 40.1g/L；Tween 80 1.0g/L；Developer 0.25g/L；Agar
15.0g/L；PH value 7.0 ± 0.2, buys biology enterococcus chromogenic culture medium rich in Qingdao sea.
(2) staphylococcus aureus basal medium：Nutriment 25g/L；Tween 80 1.0g/L；Substance that show color
40.3g/L；Agar 15.0g/L；Antipathogenic composition 6.0g/L；PH value 7.0 ± 0.2, buys biology enterococcus colour developing rich in Qingdao sea
Culture medium.
(3) chromogenic culture medium is prepared altogether for enterococcus and gold-coloured staphylococci：250mL culture medium：Enterococcus culture medium
11.28g+4.315 golden yellow grape culture medium.
(2) culture medium is weighed, heating water bath is dissolved in 250mL distilled water, when being cooled to 45-50 DEG C, sterilized petri dishes is poured into, standby
With.
4. mastitis for milk cows detection culture dish is prepared
By mastitis Gram-negative bacteria chromogenic culture medium, mastitis gram-positive bacteria chromogenic culture medium and enterococcus gold
Chromogenic culture medium is poured in the vertically dividing equally check into three culture dish of 90mm yellow grape ball respectively altogether, and marks (Fig. 1).By the inspection for preparing
Survey ware to be sealed with sealed membrane, lucifuge Cord blood (4-8 DEG C).
5. mastitis for milk cows detecting step
(1) cup son collection emulsion is adopted with aseptic；
(2) emulsion is evenly coated on the culture medium in culture dish with aseptic cotton rod；
(3) back-off culture dish, 37 DEG C are cultivated 24 hours；
(4) bacterial classification judges (with reference to table 1).
Same district bacterial classification does not judge color to table 1
| 1 area's bacterial classification | Color |
| Escherichia coli | Red |
| Klebsiella | Navy blue |
| Proteus | Brown halo |
| Pseudomonad | Butyrous, translucent |
| Candida albicans | White, opaque, petite |
| 2 area's bacterial classifications | Color |
| Streptococcusagalactiae | Blue-green |
| Streptococcus uberis | Navy blue |
| Staphylococcus aureus | Lavender carries lavender halo |
| 3 area's bacterial classifications | Color |
| Enterococcus | Redness is to purple bacterium colony |
| Staphylococcus aureus | Light blue to blue |
(5) Fig. 2 is testing result.
6 kinds of germs for causing mastitis for milk cows are detected by this method.It is Escherichia coli (redness), citric acid respectively
Bacterium (dark blue), proteus (brown halo), Streptococcusagalactiae (bluish-green), streptococcus uberis (dark blue), enterococcus (purple).
(6) this method is compared with somatic cell counting instrument
From flying crane emulsion pasture, collection emulsion is detected.Result of the test is as follows：
Body cell detector shows that cell quantity is 600,000/mL in milk, and milk cow has mammitis to infect.
This method is detected, through emulsion coated plate, is cultivated, is detected 4 kinds of bacterium (Fig. 3).Klebsiella (dark blue), agalasisa
Streptococcus (bluish-green), streptococcus uberis (dark blue), enterococcus (purple).
By comparing discovery, the simple, fast pathogenic bacteria type for detecting that mastitis for milk cows causes of this method energy, is essence
Really treatment mammitis is laid a good foundation.
(7) this method Gram-negative bacteria chromogenic culture medium compares with before improvement
Collection fresh milk is evenly coated on two kinds of culture plates, is inverted in 37 DEG C of incubators, cultivate 24 hours.A kind of
Culture plate is using basal medium (agar 15.0g/L, peptone and dusty yeast 17.0g/L, pigment 1.2g/L, the pH before improvement
Value 7.0 ± 0.2), another kind of culture plate is using the Gram-negative bacterium culture medium after present invention improvement.Result of the test is shown in Fig. 4, table
2.
After above-mentioned result of the test can be seen that the improvement of Gram-negative bacteria region, red Escherichia coli Growth is obvious.
The comparison of Gram-negative bacteria chromogenic culture medium before and after the improvement of table 2
(8) this method gram-positive bacteria chromogenic culture medium compares with before improvement
Collection fresh milk is evenly coated on two kinds of culture plates, is inverted in 37 DEG C of incubators, cultivate 24 hours.A kind of
Culture plate is using basal medium (agar 15.0g/L, peptone and dusty yeast 20.0g/L, salt 5.0g/L, the color before improvement
Plain 4.4g/L；Enriching substance：2.0g/L；PH value 6.9 ± 0.2), another kind of culture plate is using the Gram-positive after present invention improvement
Bacterium culture medium.Result of the test is shown in Fig. 5, table 3.
After above-mentioned result of the test can be seen that the improvement of gram-positive bacteria region, lavender gold-coloured staphylococci is given birth to
Long obvious.
The comparison of gram-positive bacteria chromogenic culture medium before and after the improvement of table 3
(9) Detection results of enterococcus and the common chromogenic culture medium of gold-coloured staphylococci
Collection fresh milk is evenly coated in culture plate (enterococcus of the present invention and the common chromogenic culture medium of gold-coloured staphylococci)
On, it is inverted in 37 DEG C of incubators, cultivates 24 hours.Result of the test such as Fig. 6.
As seen from Figure 6, chromogenic culture medium can detect enterococcus simultaneously altogether for enterococcus of the present invention and gold-coloured staphylococci
And staphylococcus aureus, wherein, lavender bacterium colony is enterococcus, and light blue or blue colonies are staphylococcus aureus.
Points for attention：
Ensure that coated plate is aseptic using cotton rod.
Please colony colour is judged in bright light place.
Please product low temperature (4 DEG C) is kept in dark place.
Embodiment 2
1. mastitis Gram-negative bacteria chromogenic culture medium is prepared
The present invention improvement after Gram-negative bacteria chromogenic culture medium formula be：10g/L basal medium+4g/L beef
Powder.
Wherein, basal medium is identical with the basal medium in 1 Gram-negative bacteria chromogenic culture medium of embodiment.
Preparation method is with embodiment 1.
2. mastitis gram-positive bacteria chromogenic culture medium is prepared
The present invention improvement after gram-positive bacteria chromogenic culture medium formula be：10g/L basal medium+3g/L powdered beef
+ 1g/L sodium chloride.
Wherein, basal medium is identical with the basal medium in 1 gram-positive bacteria chromogenic culture medium of embodiment.
Preparation method is with embodiment 1.
3. chromogenic culture medium is prepared altogether for enterococcus and gold-coloured staphylococci
Formula is with preparation method with embodiment 1.
4. mastitis for milk cows detection culture dish is prepared
By mastitis Gram-negative bacteria chromogenic culture medium, mastitis gram-positive bacteria chromogenic culture medium and enterococcus gold
Chromogenic culture medium is poured in the vertically dividing equally check into three culture dish of 90mm yellow grape ball respectively altogether, and marks.By the detection ware for preparing
Sealed with sealed membrane, lucifuge Cord blood (4-8 DEG C).
5. mastitis for milk cows detecting step
(1) cup son collection emulsion is adopted with aseptic；
(2) emulsion is evenly coated on the culture medium in culture dish with aseptic cotton rod；
(3) back-off culture dish, 37 DEG C are cultivated 24 hours；
(4) bacterial classification judges (with reference to table 1).
(5) result of the test is shown in Table 4,5.
The Gram-negative bacteria chromogenic culture medium testing result of the present invention of table 4
The gram-positive bacteria chromogenic culture medium testing result of the present invention of table 5
Result of the test shows that red Escherichia coli Growth is obvious on the present embodiment Gram-negative bacteria chromogenic culture medium, this
On embodiment gram-positive bacteria chromogenic culture medium, the growth of lavender gold-coloured staphylococci is obvious.
Embodiment 3
1. mastitis Gram-negative bacteria chromogenic culture medium is prepared
The present invention improvement after Gram-negative bacteria chromogenic culture medium formula be：20g/L basal medium+2g/L beef
Powder.
Wherein, basal medium is identical with the basal medium in 1 Gram-negative bacteria chromogenic culture medium of embodiment.
Preparation method is with embodiment 1.
2. mastitis gram-positive bacteria chromogenic culture medium is prepared
The present invention improvement after gram-positive bacteria chromogenic culture medium formula be：20g/L basal medium+1g/L powdered beef
+ 2g/L sodium chloride.
Wherein, basal medium is identical with the basal medium in 1 gram-positive bacteria chromogenic culture medium of embodiment.
Preparation method is with embodiment 1.
3. chromogenic culture medium is prepared altogether for enterococcus and gold-coloured staphylococci
Formula is with preparation method with embodiment 1.
4. mastitis for milk cows detection culture dish is prepared
By mastitis Gram-negative bacteria chromogenic culture medium, mastitis gram-positive bacteria chromogenic culture medium and enterococcus gold
Chromogenic culture medium is poured in the vertically dividing equally check into three culture dish of 90mm yellow grape ball respectively altogether, and marks.By the detection ware for preparing
Sealed with sealed membrane, lucifuge Cord blood (4-8 DEG C).
5. mastitis for milk cows detecting step
(1) cup son collection emulsion is adopted with aseptic；
(2) emulsion is evenly coated on the culture medium in culture dish with aseptic cotton rod；
(3) back-off culture dish, 37 DEG C are cultivated 24 hours；
(4) bacterial classification judges (with reference to table 1).
(5) result of the test is shown in Table 6,7.
The Gram-negative bacteria chromogenic culture medium testing result of the present invention of table 6
The gram-positive bacteria chromogenic culture medium testing result of the present invention of table 7
Result of the test shows that red Escherichia coli Growth is obvious on the present embodiment Gram-negative bacteria chromogenic culture medium, this
On embodiment gram-positive bacteria chromogenic culture medium, the growth of lavender gold-coloured staphylococci is obvious.
The above is only the preferred embodiment of the present invention, it is noted that for the ordinary skill people of the art
For member, under the premise without departing from the principles of the invention, some improvements and modifications can also be made, these improvements and modifications also should
It is considered as protection scope of the present invention.
Claims (8)
1. a kind of mastitis for milk cows detection kit, it is characterised in that include：Gram-negative bacteria chromogenic culture medium, gram
Positive bacteria chromogenic culture medium and the common chromogenic culture medium of enterococcus gold-coloured staphylococci；
The Gram-negative bacteria chromogenic culture medium includes first foundation culture medium and the first powdered beef base；The first foundation training
Foster base praises Gram-negative bacterium culture medium for French Kerma (unit of kinetic energy)；
The gram-positive bacteria chromogenic culture medium includes the second basal medium, the second powdered beef and sodium chloride；Described second
Basal medium praises Gram-positive bacterium culture medium for French Kerma (unit of kinetic energy)；
Chromogenic culture medium includes enterococcus basal medium and staphylococcus aureus base to the enterococcus gold-coloured staphylococci altogether
Basal culture medium.
2. mastitis for milk cows detection kit according to claim 1, it is characterised in that the Gram-negative bacteria colour developing
In culture medium, the concentration of the first foundation culture medium is 10～20g/L, and the concentration of first powdered beef is 2～4g/L；
In the gram-positive bacteria chromogenic culture medium, the concentration of second basal medium is 10～20g/L, described second
The concentration of powdered beef is 1～3g/L, and the concentration of the sodium chloride is 1～2g/L.
3. mastitis for milk cows detection kit according to claim 1 and 2, it is characterised in that the Gram-negative bacteria
In chromogenic culture medium, the concentration of the first foundation culture medium is 15g/L, and the concentration of first powdered beef is 3g/L；
In the gram-positive bacteria chromogenic culture medium, the concentration of second basal medium is 15g/L, second beef
The concentration of powder is 2g/L, and the concentration of the sodium chloride is 1.5g/L.
4. mastitis for milk cows detection kit according to any one of claim 1 to 3, it is characterised in that the leather is blue
The pH value of family name's negative bacterium chromogenic culture medium is 7.0 ± 0.2, the pH value of the gram-positive bacteria chromogenic culture medium is 6.9 ±
0.2, the pH value of the common chromogenic culture medium of the enterococcus gold-coloured staphylococci is 7.0 ± 0.2.
5. mastitis for milk cows detection kit according to any one of claim 1 to 4, it is characterised in that with weight portion
Meter, the enterococcus basal medium include：40.1 parts of nutriment, 1 part of Tween 80,0.25 part of developer, 15 parts of agar.
6. mastitis for milk cows detection kit according to any one of claim 1 to 5, it is characterised in that with weight portion
Meter, the staphylococcus aureus basal medium include：25 parts of nutriment, 1 part of Tween 80,40.3 parts of substance that show color, fine jade
15 parts of fat, 6 parts of antipathogenic composition.
7. mastitis for milk cows detection kit according to any one of claim 1 to 6, it is characterised in that the intestines ball
Bacterium basal medium is (10～15) with the mass ratio of the staphylococcus aureus basal medium：(1～5).
8. mastitis for milk cows detection kit according to any one of claim 1 to 7, it is characterised in that the milk cow
Mammitis detection kit also includes vertically dividing equally check into three culture dish.
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| CN108192947A (en) * | 2018-01-23 | 2018-06-22 | 杨海龙 | The method of mastitis for milk cows pathogen identification kit and pathogen identification |
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