CN106480161B - 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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- CN106480161B CN106480161B CN201611009459.3A CN201611009459A CN106480161B CN 106480161 B CN106480161 B CN 106480161B CN 201611009459 A CN201611009459 A CN 201611009459A CN 106480161 B CN106480161 B CN 106480161B
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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.Mastitis for milk cows detection kit provided by the invention can quick and precisely detect recessive type mastitis for milk cows, it is not necessary to by specific apparatus, the direct judged result of naked eyes can, technical staff is easy to grasp, easy to operate, and result can be gone out in 18h~24h, simultaneously can be low with batch detection, 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 caused by the stimulation of the factors such as physics, chemistry, microorganism
The inflammation of cow breast, show as milk and the change of physicochemical property and cytology property, breast tissue generation pathology change occurs
Change, be one of regular incidence of milk cow.Mammitis is divided into clinical type breast according to breast and milk whether there is clinical visible change
Scorching (Clinical Mastitis) and subclinical mastitis (Subclinical Mastitis).Recessive mastitis shows as suffering from
The breast and milk of ox visually observe without exception, it is necessary to by Somatic Cell Count or by the detectable breast of other diagnostic methods
Room inflammation.
Counted according to international milk cow federation, in the 1970s, milk cow clinic mastitis illness rate about 2%, recessive breast
Room inflammation infected cattle is up to 50%；In 11,000,000 cow heads of U.S.'s raising, half suffers from all kinds mammitis；The Japanese 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 of 32, the city in South China 22 cattle farm 1037
The room inflammation cause of disease and incidence investigation, and wherein 5 20, area cows in milk of cattle farm 3384 are tested with Lanzhou mammitis
Carry out recessive mastitis investigation, the results showed 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 summary, mastitis for milk cows is in world wide
Interior infection rate is all very high, and in China, the infection conditions of mastitis for milk cows are also quite serious, and 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 external conventional estimated to economic loss caused by mastitis for milk cows is the drop according to milk yield
The increase of low, milk loss, medication expense, animal doctor's expense and labour's expense etc. counts.It is reported that milk is treated every year by Britain
The average cost of garget is per 187 pounds of cow head, and it is 177-182 that the U.S. is used for per the cost of 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 generation of mammitis is huge to the economic loss that milk cow production is brought.In these economic losses, due to the reduction of the output of milk
Caused loss accounts for 70%, milk cow is eliminated ahead of time due to ill and accounts for 14%, and discarded milk accounts for 7%, treatment and animal doctor's expense
8% etc. is accounted for, and economic loss mostlys come from recessive mastitis, and loss caused by it accounts for the annual total productive capacity of cow
10-11%.Secondly, mammitis is greatly reduced the quality of breast, containing abundant 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 be damaged, and some nutritional ingredients are lost, and make nutrition
Composition is not complete and reduces the health-care effect of milk.Total amino acid content is also by average 2.2387g/100mL in recessive mastitis milk
Reduce 21%, wherein methionine, isoleucine, leucine, phenylalanine, 5 kinds of essential amino acids of lysine and histidine, essence
Propylhomoserin significantly reduces；Lactose, casein, fat, Ca2+、P3-、K+Content Deng beneficiating ingredient is reduced or substantially reduced；And exempt from
The content of the harmful components such as epidemic disease globulin, esterase, inflammatory factor, pathogenic bacteria and its caused toxin then increases, heat endurance drop
It is low.In addition, milk cow, which is suffered from the milk produced after mammitis disease, contains substantial amounts of inflammatory factor, pathogenic bacteria and its caused poison
Element, sterilize not tight or mixed up the milk containing pathogenic bacteria if drunk, can make one to feel not quite the thing, severe patient can lure
Send out disease.There is abuse of antibiotics in mammitis lysis is treated, some operators, which are not eliminated during treating, suffers from cow's milk
Juice, antibiotic being caused in Residues in Milk, causing eater that allergic reaction occurs, especially the elderly and infant are endangered can more
Greatly.Therefore, it is particularly important that diagnosis is made to mastitis for milk cows disease in time, exactly.
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 monitor at any time, make in time
Diagnosis, so that cluster prevents.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.,
It is commonly used at present for body cell direct counting method at home.Pathological research confirms, a large amount of white thin after breast infection inflammation
Born of the same parents enter mammary gland, part epithelial cell shedding, somatic number in milk is significantly raised.According to this principle, every milliliter is calculated
Somatic number in milk, this method testing result is the most accurate, and this is the benchmark for diagnosing recessive mastitis, and other diagnosis sides
The standard that method compares.Indirect method is clinically the most frequently used method at present, and it is divided into chemical measure and physical measure, its
In at present it is clinically the most commonly used with chemical measure.Chemical measure is indirect determination latex cell number and milk pH value
Method.Room is scorching when occurring, and the pH value of milk rises, can be to reach the purpose of judgement by determining milk pH value.But these are examined
Survey method is required to special detecting instrument, complex operation, requires high to operating personnel.A kind of accordingly, it is desirable to provide operation letter
List, fast and accurately mastitis for milk cows detection method.
The 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 which, by specific apparatus, the direct judged result of naked eyes can, 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 following technical scheme：
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 is that French Kerma (unit of kinetic energy) praises Gram-positive bacterium culture medium；
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 embodiment provided by the invention, 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 embodiment provided by the invention, 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 embodiment provided by the invention, 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 is bought in the rich biological enterococcus colour developing culture in Qingdao sea
Base.
In embodiment provided by the invention, 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, the mass ratio of enterococcus basal medium and staphylococcus aureus basal medium is (11~12)：
(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 bacteriums 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, mastitis for milk cows detection kit provided by the invention can quick and precisely detect recessive type mastitis for milk cows, be not required to
Will be by specific apparatus, the direct judged result of naked eyes can, technical staff is easy to grasp, easy to operate, in 18h~24h
Result can be gone out, while can be low with batch detection, cost；
2nd, in mastitis for milk cows detection kit provided by the invention, Gram-negative bacteria chromogenic culture medium and gram sun
Property bacterium chromogenic culture medium can remarkably promote the growth of bacterium, can shorten detection time after improvement；
3rd, in the common chromogenic culture medium of enterococcus gold-coloured staphylococci, enterococcus basal medium and staphylococcus aureus base
After basal culture medium combines in specific proportions, enterococcus and staphylococcus aureus can be detected simultaneously.
Brief description of the drawings
Fig. 1 show the present invention prepare include mastitis Gram-negative bacteria chromogenic culture medium, mastitis gram-positive bacteria
The vertically dividing equally check into three culture dish of chromogenic culture medium and the common chromogenic culture medium of enterococcus Staphylococcus aureus；
Fig. 2 shows the Pathogen detection result to mastitis for milk cows using culture medium of the present invention；
Fig. 3 shows the testing result of the emulsion using the pasture collection of culture medium of the present invention detection flying crane emulsion；
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.
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.In particular, all similar replacements and change come to those skilled in the art
Say it is it will be apparent that they are considered as being included in the present invention.The method of the present invention and application have passed through preferred embodiment
Be described, related personnel substantially can not depart from present invention, in spirit and scope to method described herein and should
With being modified or suitably change with combining, to realize and using the technology of the present invention.
Term is explained：
1st, mastitis for milk cows
Cow breast is immersed during galactopoiesis by mechanical irritation, pathogenic microorganism and the damage of chemicals rationality is drawn
The breast pathological change risen, is divided into serosity mammitis, cellulosic catarrh mammitis, suppurative mammitis, hemorrhagic breast
Scorching, gangrenous mastitis and recessive mastitis.
2nd, pathogenic infection
The infection as caused by parasite or microorganism, so far, people have isolated 150 from cow mammary gland tissue
Kind pathogenic microorganism, it is most common to have 7 kinds of 23 kinds, wherein 14 kinds of bacterium, 2 kinds of mycoplasma, fungi and 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
For the nutrient matrix of microbial growth.It is divided into fluid nutrient medium and solid medium.Typically, in Liquid Culture
Base adds agar Solid agar culture is made.Culture medium has different formulas typically all to contain carbon source, nitrogen source and inorganic salts.
Used medium raw material can be bought by market in mastitis for milk cows detection kit provided by the invention.
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 and praise Gram-negative bacterium culture medium in French Kerma (unit of kinetic energy).
Preparation method is as follows：
(1) basal medium and powdered beef are weighed by above-mentioned formula, be dissolved in the clean triangular flask of 1000mL deionized waters.
Also the amount for preparing culture medium can be proportionally expanded or shunk as needed.
(2) 100 DEG C are heated to, does not stop to stir, is completely dissolved it.It is sure not to be heated beyond 100 DEG C.If added using micro-wave oven
Heat, culture medium ebuillition of heated should immediately be removed, gently shaken up, place into microwave stove heat, observation minute bubbles are changed into air
Bubble, until being completely dissolved.It is sure not to overflow culture medium.
(3) culture medium dissolved is cooled to 45 DEG C~50 DEG C, gently mixed, pour plate, make its solidification, dried 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 beefs
+ 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, buy and praise 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 waters
In the bottle of angle, mixing is sufficiently stirred.Also the amount for preparing culture medium can be proportionally expanded or shunk as needed.
(2) 100 DEG C are heated to, does not stop to stir, is completely dissolved it.It is sure not to be heated beyond 100 DEG C.If added using micro-wave oven
Heat, culture medium ebuillition of heated should immediately be removed, gently shaken up, place into microwave stove heat, observation minute bubbles are changed into air
Bubble, until being completely dissolved.It is sure not to overflow culture medium.
(3) culture medium dissolved is cooled to 45 DEG C~50 DEG C, gently mixed, pour plate, make its solidification, dried 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, buy and win biological enterococcus chromogenic culture medium 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, buy in the rich biological enterococcus colour developing in Qingdao sea
Culture medium.
(3) chromogenic culture medium is prepared altogether for enterococcus and gold-coloured staphylococci：250mL culture mediums：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, is poured into sterilized petri dishes, standby
With.
4. prepare mastitis for milk cows detection culture dish
By mastitis Gram-negative bacteria chromogenic culture medium, mastitis gram-positive bacteria chromogenic culture medium and enterococcus gold
Chromogenic culture medium is poured in 90mm vertically dividing equally check into three culture dish yellow grape ball respectively altogether, and marks (Fig. 1).The inspection that will be prepared
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 sterile；
(2) emulsion is evenly coated on the culture medium in culture dish with sterile cotton rod；
(3) back-off culture dish, 37 DEG C are cultivated 24 hours；
(4) strain judges (with reference to table 1).
Same district strain does not judge color to table 1
| 1 area's strain | Color |
| Escherichia coli | It is red |
| Klebsiella | Navy blue |
| Proteus | Brown halo |
| Pseudomonad | Butyrous, it is translucent |
| Candida albicans | White, opaque, petite |
| 2 area's strains | Color |
| Streptococcusagalactiae | Blue-green |
| Streptococcus uberis | Navy blue |
| Staphylococcus aureus | Lavender carries lavender halo |
| 3 area's strains | Color |
| Enterococcus | It is red to purple bacterium colony |
| Staphylococcus aureus | It is 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 (red), 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 infection.
This method detects, and by emulsion coated plate, culture, detects bacterium 4 kinds (Fig. 3).Klebsiella (dark blue), agalasisa
Streptococcus (bluish-green), streptococcus uberis (dark blue), enterococcus (purple).
By it was found that, this method can it is simple, fast detect pathogenic bacteria type caused by mastitis for milk cows, for essence
Really treatment mammitis is laid a good foundation.
(7) this method Gram-negative bacteria chromogenic culture medium is compared with before improveing
Collection fresh milk is evenly coated on two kinds of culture plates, is inverted in 37 DEG C of incubators, is cultivated 24 hours.It is a kind of
Culture plate is using basal medium (agar 15.0g/L, peptone and dusty yeast 17.0g/L, the pigment 1.2g/L, pH before improvement
Value 7.0 ± 0.2), another 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 can be seen that the improvement of Gram-negative bacteria region by above-mentioned result of the test, red Escherichia coli Growth is obvious.
Table 2 improves the comparison of front and rear Gram-negative bacteria chromogenic culture medium
(8) this method gram-positive bacteria chromogenic culture medium is compared with before improveing
Collection fresh milk is evenly coated on two kinds of culture plates, is inverted in 37 DEG C of incubators, is cultivated 24 hours.It is 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 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 the improvement of gram-positive bacteria region being can be seen that by above-mentioned result of the test, the life of lavender gold-coloured staphylococci
It is long obvious.
Table 3 improves the comparison of front and rear gram-positive bacteria chromogenic culture medium
(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：
It is sterile to ensure that coated plate uses cotton rod.
Please judge colony colour 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 the Gram-negative bacteria chromogenic culture medium of embodiment 1.
Preparation method is the same as 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 beefs
+ 1g/L sodium chloride.
Wherein, basal medium is identical with the basal medium in the gram-positive bacteria chromogenic culture medium of embodiment 1.
Preparation method is the same as embodiment 1.
3. chromogenic culture medium is prepared altogether for enterococcus and gold-coloured staphylococci
Formula is with preparation method with embodiment 1.
4. prepare mastitis for milk cows detection culture dish
By mastitis Gram-negative bacteria chromogenic culture medium, mastitis gram-positive bacteria chromogenic culture medium and enterococcus gold
Chromogenic culture medium is poured in 90mm vertically dividing equally check into three culture dish yellow grape ball respectively altogether, and is marked.The detection ware that will be prepared
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 sterile；
(2) emulsion is evenly coated on the culture medium in culture dish with sterile cotton rod；
(3) back-off culture dish, 37 DEG C are cultivated 24 hours；
(4) strain 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
The growth of lavender gold-coloured staphylococci is obvious on embodiment gram-positive bacteria chromogenic culture medium.
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 the Gram-negative bacteria chromogenic culture medium of embodiment 1.
Preparation method is the same as 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 beefs
+ 2g/L sodium chloride.
Wherein, basal medium is identical with the basal medium in the gram-positive bacteria chromogenic culture medium of embodiment 1.
Preparation method is the same as embodiment 1.
3. chromogenic culture medium is prepared altogether for enterococcus and gold-coloured staphylococci
Formula is with preparation method with embodiment 1.
4. prepare mastitis for milk cows detection culture dish
By mastitis Gram-negative bacteria chromogenic culture medium, mastitis gram-positive bacteria chromogenic culture medium and enterococcus gold
Chromogenic culture medium is poured in 90mm vertically dividing equally check into three culture dish yellow grape ball respectively altogether, and is marked.The detection ware that will be prepared
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 sterile；
(2) emulsion is evenly coated on the culture medium in culture dish with sterile cotton rod；
(3) back-off culture dish, 37 DEG C are cultivated 24 hours；
(4) strain 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
The growth of lavender gold-coloured staphylococci is obvious on embodiment gram-positive bacteria chromogenic culture medium.
Described 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 (4)
- A kind of 1. mastitis for milk cows detection kit, it is characterised in that including：Gram-negative bacteria chromogenic culture medium, gram Positive bacteria chromogenic culture medium and the common chromogenic culture medium of enterococcus staphylococcus aureus；The Gram-negative bacteria chromogenic culture medium includes first foundation culture medium and the first powdered beef；The first foundation culture Base is that French Kerma (unit of kinetic energy) praises Gram-negative bacterium culture medium；The gram-positive bacteria chromogenic culture medium includes the second basal medium, the second powdered beef and sodium chloride；Described second Basal medium is that French Kerma (unit of kinetic energy) praises Gram-positive bacterium culture medium；Chromogenic culture medium includes enterococcus basal medium and staphylococcus aureus to the enterococcus staphylococcus aureus altogether Basal medium；In the Gram-negative bacteria chromogenic culture medium, the concentration of the first foundation culture medium is 10 ~ 20g/L, described first The concentration of 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；The enterococcus basal medium includes：40.1 parts of nutriment, 1 part of Tween 80,0.25 part of developer, agar 15 Part；The staphylococcus aureus basal medium includes：25 parts of nutriment, 1 part of Tween 80,40.3 parts of substance that show color, 15 parts of agar, 6 parts of antipathogenic composition；The mass ratio of the enterococcus basal medium and the staphylococcus aureus basal medium is（10~15）：（1~ 5）.
- 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 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.
- 3. mastitis for milk cows detection kit according to claim 1 or 2, it is characterised in that the Gram-negative bacteria The pH value of chromogenic culture medium is 7.0 ± 0.2, and the pH value of the gram-positive bacteria chromogenic culture medium is 6.9 ± 0.2, the intestines The pH value of the common chromogenic culture medium of coccus staphylococcus aureus is 7.0 ± 0.2.
- 4. mastitis for milk cows detection kit according to claim 1 or 2, it is characterised in that the mastitis for milk cows inspection Test agent box also includes vertically dividing equally check into three culture dish.
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