CN1858233A - Preparing method for cow mammitis streptococcus culture fluid and its use - Google Patents
Preparing method for cow mammitis streptococcus culture fluid and its use Download PDFInfo
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Abstract
The preparation process of cow mammitis streptococcus culture fluid and its use belongs to the field of animal epidemic disease diagnosing, preventing and treating technology. The preparation process includes dissolving peptone 10 g, yeast powder 5 g, sodium chloride 10 g, glucose 5 g and sodium azide 0.5 g in water of 1 L; regulating pH to 7.2 with sodium hydroxide; adding bacteria-free 0.1% concentration crystal violet water solution in 1.3ml and 1% concentration bromocreasol purple water solution in 1.5ml; sterilizing at 121deg.c and preservation at 4 deg. c. The culture fluid may be used in the fast identification of cow mammitis streptococcus and antibiotic sensitivity test.
Description
Technical field
This invention belongs to animal epidemic diagnosis and prevention and control field.
Technical background
Mammitis of cow sickness rate height, hazardness is big, is one of main disease that influences world's dairy development.According to the data record of international milk federation, nearly 50% cow suffers from mastitis, and wherein nearly 2% suffers from clinic mastitis, and 48% suffers from latent mammitis.China's clinic mastitis sickness rate 33.41% (9.7%～55.6%), the recall rate of latent mammitis are 50-80%.Clinic mastitis has lactation amount decline, mammary swelling, milk significantly clinical symptom such as rotten, though and the breast of latent mammitis and milk outward appearance do not have the visible naked eyes and change, usually cause the quality of raw material milk to descend, threaten that people's is healthy.According to estimates, China every year because of financial loss that mastitis caused above 3.16 hundred million yuan.The pathogenic micro-organism that causes mastitis mainly comprises gold-coloured staphylococci, streptococcus agalactiae, streptococcus dysgalactiae, cow's milk room suis, intestinal bacteria, klebsiella, Pseudomonas aeruginosa and some other fungi, virus etc.But in these pathogenic agent,, account for whole mammitis of cow case more than 90% based on streptococcus aureus, suis and intestinal bacteria.
The treatment of mammitis of cow is based on microbiotic, but because antibiotic use lack of standardization has caused serious problems such as antibiotic remains in the milk, Resistant strain is general, the mastitis that pasts medical help case increases.The major cause that causes this phenomenon is because the restriction of diary farm economy, technology and experiment condition, seldom carrying out necessary bacterium before with antibiotic control mammitis of cow separates and drug sensitivity test, only depend on animal doctor's hobby and experience, long-term, blindly and excess use antibiotic.
Domestic and international normally used drug sensitive test method mainly contains disk diffusion method, broth dilution method, agar dilution, automatization susceptibility instrument method and Etest method at present.Wherein, disk diffusion method is the responsive or tolerance of judging bacterial antibiotic by the diameter of measuring inhibition zone, great advantage is that expense is low and medicament selection is flexible, inoculated multiple factor affecting such as bacterium amount, incubation time, antibiotic content and diffusive force, plate thickness but exist, need strictness to carry out stdn, just can guarantee result's reliability, and have consuming time, effort and shortcomings such as inapplicable anerobe and severe oxygen bacterium; Broth dilution method and agar dilution are the methods of the microbiotic doubling dilution being come quantitative assay minimum inhibitory concentration (MIC) in meat soup or agar, its advantage is can accurately measure some anerobes and severe oxygen bacterium to antibiotic susceptibility, and one the cover plate can survey many strains bacterium, but have shortcoming such as consuming time and effort; Automatization susceptibility instrument is to come sentence read result by detecting turbidity, the fluorescence intensity of fluorescent indicator or the hydrolysis reaction of fluorogenic substrate, have quick and advantage such as powerful function of statistic analysis, lack handiness, be difficult to detect shortcomings such as some special bacterium and instrument cost an arm and a leg but have medicament selection; The Etest method is in conjunction with the advantage of diffusion process and dilution method, with the susceptibility that contains continuous concentration gradient microbiotic plastic strip bacterial detection, has accurately, advantage such as reliable, good stability, can be used for the drug sensitive test of various bacteria, and unique shortcoming is to cost an arm and a leg.
Evaluation and drug sensitive test as for cow mammitis streptococcus, method commonly used both at home and abroad at present is the milk sample inoculation sheep blood agar plate with aseptic collection, cultivate after 18-48 hour for 37 ℃, according to colonial morphology, haemolysis characteristics and gram stain microscopy etc. are made preliminary evaluation, the suspicious bacterium colony of picking carries out biochemical tests such as CAMP test and Vitamin C2 hydrolysis and confirms again, on the Mueller-Hinton that contains 5% sheep blood agar plates such as (MH), carry out the disk diffusion method drug sensitive test with colonies typical at last, or microbiotic is diluted in the mensuration of carrying out minimal inhibitory concentration (MIC) in the meat soup.These methods not only needed just can obtain a result at least in 3-5 days, possessed equipment, reagent and technical qualification that necessary bacteriology is identified, and can not truly reflect sometimes that with the drug sensitive test result that single bacterium colony carries out bacterium " colony " is to antibiotic susceptibility.As for automatization susceptibility instrument and Etest method, be difficult to by domestic veterinary clinic and diary farm acceptance because price is very expensive.
Summary of the invention
Purpose of the object of the invention provides the spread cultivation compound method of milk milk sample streptococcus intermedius nutrient solution of a kind of fast selective.
Characteristics of the present invention are: 10g peptone, 5g yeast powder, 10g sodium-chlor, 5g glucose and 0.5g sodium azide are dissolved in the 1L water, adjust pH to 7.2 with sodium hydroxide solution, add aseptic 0.1% Viola crystallina aqueous solution 1.3ml and 1% purpurum bromocresolis aqueous solution 1.5ml again, 121 ℃ of sterilizations down, standby in 4 ℃ of preservations at last.
Also can in above-mentioned every liter of liquid medium, add the 15g agar powder and make the EN agar plate.
Another purpose provides the purposes of above-mentioned cow mammitis streptococcus culture fluid among the present invention:
One of purposes is: the method that is used to differentiate cow mammitis streptococcus: inoculation milk sample in containing the cultivation plate hole of nutrient solution, 37 ℃ cultivate 24 hours down after, nutrient solution becomes yellow by purple and is judged as in the milk sample suis is arranged.
Two of purposes is: be used for the drug sensitive test of cow mammitis streptococcus: nutrient solution is sub-packed in is coated with antibiotic culture plate, the inoculation suis is selected culture or the positive milk of mastitis sample for the first time, cultivated 24 hours down at 35 ℃, nutrient solution is to become yellow by purple to be judged as microbiotic is had susceptibility.
The present invention is on LB nutrient solution basis, add the calf serum and glucose, the sodium azide of inhibition gram negative bacterium growth, the Viola crystallina of the non-suis Gram-positive bacteria growing of inhibition and the purpurum bromocresolis that indication suis glucose fermentation produces acid that promote streptococcus growth, the nutrient solution that adopts the inventive method to make is only selected to spread cultivation to the suis in the mammitis of cow milk sample, and the speed that spreads cultivation is fast.
When having or not suis in judging the milk sample, after cultivating 24 hours, nutrient solution is judged as in the milk sample suis is arranged as become yellow by purple; As nutrient solution still is purple, then, does not have suis in the milk sample.
When measuring suis to antibiotic susceptibility, after cultivating 24 hours, become yellow as nutrient solution by purple and be judged as corresponding microbiotic is had susceptibility, still be purple as nutrient solution, then, corresponding microbiotic is had resistance.
The present invention is on the basis of existing LB nutrient solution, develop and can be directly carry out suis and differentiate fast and cultivate and the drug sensitive test method with clinical milk sample, its remarkable advantage is quick, accurate, inexpensive and does not need complicated plant and instrument, is particularly suitable for domestic middle-size and small-size diary farm and the animal doctor of basic unit and uses.
Description of drawings
Fig. 1 is the growth curve chart of suis in EN nutrient solution and serum nutrient broth.
Specific embodiment
One, preparation EN nutrient solution:
Take by weighing peptone 10g, yeast powder 5g, sodium-chlor 10g, glucose 5g, sodium azide 0.5g respectively with balance, add in the 1L water, stirring makes each composition water-soluble, adjust pH value to 7.2 with sodium hydroxide solution again, add filtration sterilization 0.1% Viola crystallina aqueous solution 1.3ml and 1% purpurum bromocresolis aqueous solution 1.5ml again, sterilization is 15 minutes under 121 ℃ of high temperature, makes liquid EN nutrient solution, preserve down at 4 ℃, standby.
In above-mentioned every liter of liquid EN nutrient solution, add 15 gram agar powders and be the EN agar plate.Preserve down at 4 ℃, standby.
Two, suis is differentiated fast:
Judge tentatively according to clinical examination or California mastitis test (CMT) detected result and to suffer from clinical type or latent mammitis milk cow, squeeze with conventional method of milking and remove first three milk that usefulness routine disinfection liquid disinfectant nipple and near zone thereof are with the cotton ball soaked in alcohol nipple of sterilizing.The sterilised test tube lid is opened, on the other hand test tube is kept horizontality, another hand milking, and milk " is injected " in the test tube, and build the test tube lid immediately, place ice bath to take back real (inspection) immediately and test the chamber.In aseptic worktable, open 24 or 96 aseptic well culture plates, the EN nutrient solution is sub-packed in the cultivation plate hole, every hole 200 μ l, in every hole, add undiluted or with the suitable positive milk of the mastitis sample 10 μ l of dilution of physiological saline, build back 37 ℃ and leave standstill cultivation 24 hours, whether become yellow judgement according to nutrient solution and have or not streptococcus growth by purple.
Three, quick medicine-sensitive test:
1, determining and culture plate bag quilt of microbiotic minimal inhibitory concentration: the criteria for interpretation and relevant minimal inhibitory concentration (MIC) that suppress circle with reference to the stdn council of international standard U.S. clinical labororatory (NCCLS20012) about streptococcus drug sensitive, filter out 8 kinds of antibiotic sensitivities of 7 classes or Resistant strain, with 0.5,1.0,1.5 and 2.0 times of 8 kinds of antibiotic bags of 7 classes to standard MIC are by 96 well culture plates, with EN is that nutrient solution carries out streptococcic quick medicine-sensitive test, by comparing, determine that the EN nutrient solution is to the influence of examination microbiotic bacteriostatic activity and suitable microbiotic culture plate package amount with standard disk diffusion method drug sensitive test result.Microbiotic is made into 1000 times of MIC concentration after proofreading and correct, packing 96 well culture plates, in aseptic worktable, dry up naturally or lyophilize after, 4 ℃ of preservations are standby;
2, quick medicine-sensitive is tested: the EN nutrient solution is sub-packed in is coated with antibiotic 96 well culture plates, the inoculation suis is selected culture or the positive milk of mastitis sample for the first time, cultivated 24 hours for 35 ℃, whether become yellow judgement according to nutrient solution and have or not streptococcus growth to reach corresponding antibiotic susceptibility by purple.
Four, simultaneous test:
1, the growing state of suis on the EN agar plate: will suffer from the isolating streptococcus agalactiae of ox, streptococcus dysgalactiae, streptococcus faecium streak inoculation EN agar plate from mastitis, cultivate after 24 hours for 37 ℃, three kinds of streptococcus growths become typical suis bacterium colony, the about 1mm of colony diameter, neat in edge, tiny small colonies as tip-like, the picking colonies typical is made nacterial smear, microscopy behind gramstaining, the suis that grows on the EN agar plate is a Gram-positive, is typical catenation.The picking colonies typical carries out biochemical test, and the streptococcus agalactiae CAMP test is positive, and Vitamin C2 hydrolysis and the test of anti-sodium-chlor are negative; Streptococcus dysgalactiae CAMP test and the test of anti-sodium-chlor are negative, and the Vitamin C2 hydrolysis experiment is suspicious; Streptococcus faecium CAMP test feminine gender, Vitamin C2 hydrolysis and the test of anti-sodium-chlor are positive.
2, the growth curve of suis in the EN nutrient solution: picking is cultivated the single bacterium colony of streptococcus agalactiae, streptococcus dysgalactiae and streptococcus faecium on blood agar plate, inoculation contains the nutrient broth of 5% calf serum respectively, cultivate after 12 hours for 37 ℃, ratio inoculation 2mlEN nutrient solution and serum nutrient broth in 1: 100, every kind of nutrient solution inoculation 12 pipes, in 37 ℃ of shaking culture processes, take out 1 pipe every 2 hours and place 4 ℃ of refrigerators, when in the end a pipe takes out, unified OD with each pipe of spectrophotometric determination
600Value is with OD
600Be ordinate zou, incubation time is an X-coordinate, traces the growth curve of three kinds of suis in two kinds of liquid nutrient mediums, as shown in Figure 1.Ordinate zou is the absorbance value of bacterial cultures among Fig. 1, and X-coordinate is an incubation time.
3, the selectivity characteristic of EN nutrient solution: get suis and (comprise streptococcus faecium, streptococcus agalactiae, streptococcus dysgalactiae), non-suis gram-positive microorganism (comprises streptococcus aureus, staphylococcus epidermidis, Bacillus subtilus and bacillus rhusiopathiae suis), Gram-negative bacteria (comprises intestinal bacteria, Salmonella typhimurium, enteroaerogen, avian pasteurella multocida, dysentery bacterium, false unit cell Pseudomonas aeruginosa, Bacillus proteus) broth culture 20 μ l, inoculate 2ml EN nutrient solution respectively, 37 ℃ leave standstill cultivate 24h after, streptococcus faecium, streptococcus agalactiae and streptococcus dysgalactiae be well-grown in the EN nutrient solution, and the nutrient solution color becomes yellow by purple; Gram negative bacteriums such as non-suis gram-positive microorganism such as streptococcus aureus, staphylococcus epidermidis, Bacillus subtilus, bacillus rhusiopathiae suis and intestinal bacteria, Salmonellas, enteroaerogen, pasteurellosis bacillus, dysentery bacterium, false unit cell Pseudomonas aeruginosa, Bacillus proteus do not grow, the purple of nutrient solution does not become, and shows that the EN nutrient solution has good selectivity characteristic to suis.
Get streptococcus agalactiae, streptococcus dysgalactiae and streptococcus faecium serum nutrient broth culture 20 μ l respectively, mix with the streptococcus aureus and the intestinal bacteria broth culture of equivalent, 6ml EN nutrient solution is gone in inoculation, 37 ℃ leave standstill cultivate 24h after, respectively get culture 50 μ l, inoculate blood agar plate, Baird-Parker selection agar plate and maconkey agar flat board respectively, after 24h is cultivated in 37 ℃ of continuation, only on the blood agar plate streptococcus growth is arranged, no staphylococcus and intestinal bacteria growth on Baird-Parker agar plate and maconkey agar flat board.
4, the EN nutrient solution is to streptococcic selection sensitivity: single colony inoculation EN selects nutrient solution with known streptococcus agalactiae, streptococcus dysgalactiae and streptococcus faecium, cultivates after 12 hours, carries out bacterial count with colony counting method for 37 ℃.With the EN nutrient solution suis culture is done 10
1, 10
2, 10
3, 10
4, 10
5, 10
6, 10
7, 10
8, 10
9, 10
10Doubly dilution, every extent of dilution is got 20 μ l inoculation 2ml EN selectivity nutrient solution, cultivate after 24 hours for 37 ℃, preliminary judgement has or not streptococcus growth according to the nutrient solution colour-change, gets 100 μ l cultures coating blood agar plate then from every pipe, counts bacterium colony behind 37 ℃ of cultivation 24h, with the highly diluted multiple that streptococcus growth is arranged is that sensitivity is cultivated in the selection of EN nutrient solution, the selection sensitivity of streptococcus agalactiae as a result is 35 bacteriums, and streptococcus dysgalactiae is 15 bacteriums, and streptococcus faecium is 25 bacteriums.
5, the EN nutrient solution selects to cultivate the effect of clinical milk sample streptococcus intermedius: detect (CMT method) result according to clinical diagnosis and milk sample somatocyte, 18 parts in clinical type of aseptic collection and recessive mammitis of cow milk sample are defined as streptococcal infection milk sample with conventional blood agar plate partition method and biochemical test.The sample of will suckling is suitably inoculated EN nutrient solution, blood agar plate, Baird-Parker agar plate, maconkey agar flat board respectively after the dilution, cultivate after 24 hours for 37 ℃, in the EN nutrient solution of 18 parts of milk samples inoculation streptococcus growth is arranged all, the nutrient solution color becomes yellow by purple; On the blood agar plate of 18 parts of milk samples inoculation typical suis bacterium colony is arranged all, grow golden yellow staphylococcus and intestinal bacteria bacterium colony (seeing Table 1) on the part blood agar, the declaratives milk cow is the polyinfection of two or three bacterium.
6, the EN nutrient solution to the influence of microbiotic bacteriostatic activity and suitable microbiotic bag determining by concentration: use suis 32 strains that from clinical milk sample, are separated to, suppress the explanation and relevant minimal inhibitory concentration (MIC of circle about the streptococcus susceptibility with reference to NCCLS20012, see Table 2), carry out the disk diffusion method drug sensitive test containing on the MH substratum of 5% calf serum, determine that suis is to 8 kinds of antibiotic susceptibility of 7 classes (seeing Table 3).The single bacterium colony of picking suis is inoculated 2ml serum nutrient broth respectively, cultivates after 12 hours for 37 ℃ and uses for the quick medicine-sensitive test.With EN nutrient solution packing 96 well culture plates, every hole 200 μ l, 0.5,1.0,1.5 or 2.0 times of antibiotic to standard MIC is added in the respective aperture, 10 μ l suis cultures are inoculated in every hole, cultivate after 24 hours for 35 ℃, whether become yellow according to nutrient solution, judge to have or not streptococcus growth and corresponding antibiotic susceptibility by purple.As shown in table 3, the EN nutrient solution is to the not influence of the bacteriostatic activity of penicillin, tsiklomitsin and gentamicin, and the bacteriostatic activity of ceftriaxone, Ciprofloxacin, clindamycin, erythromycin, Ofloxacine USP 23 is had certain influence.In view of the above, when carrying out the quick medicine-sensitive test with the EN nutrient solution, by the concentration of ceftriaxone, Ciprofloxacin, clindamycin, Ofloxacine USP 23, erythromycin is made suitably to adjust (table 4), the drug sensitive test result of acquisition is consistent with the standard disk diffusion method.
7, carry out the quick medicine-sensitive test with the positive milk of mastitis sample: in aseptic worktable, the EN nutrient solution is sub-packed in is coated with antibiotic 96 well culture plates, every hole 200 μ l, in each hole, add 10 μ l suis respectively and select culture or the positive milk of mastitis sample for the first time, cultivate after 24 hours for 35 ℃, whether become yellow judgement according to nutrient solution and have or not streptococcus growth and whether responsive corresponding microbiotic by purple.The result shows, the quick medicine-sensitive test-results that to be nutrient solution with EN carry out 8 parts of suis positive milk samples and suis pure culture thereof and the disk diffusion method identical (seeing Table 5) of standard.
Above result of implementation shows that streptococcic EN nutrient solution (base) has very strong selection specificity, sensitivity and distinguishing ability to cow mammitis streptococcus.With EN nutrient solution and 24 or 96 hole microbial culture plate combinations, the method for quick identification of the cow mammitis staphylococcus of setting up, whether as long as nutrient solution is sub-packed in cultivates plate hole, inoculation trace milk sample and cultivate through 24 hours, can clearly draw is the conclusion of streptococcal infection.To be coated with and proofread and correct back antibiotic 96 well culture plates of MIC and the combination of EN nutrient solution, the quick medicine-sensitive test method of the cow mammitis streptococcus of setting up, as long as, can know clearly that suis is to which kind of antibiotic sensitive or resistance with nutrient solution packing, inoculation micro-example and cultivation in 24 hours.Quick discriminating of the suis of setting up and drug sensitive test not only can be used for the drug sensitive test of suis pure culture, the more important thing is and directly to carry out drug sensitive test with clinical milk sample, its outstanding advantage is quick, easy, inexpensive and does not need specialized equipment equipment, is particularly suitable for diary farm and clinical animal doctor and uses.
Penicillin, tsiklomitsin and the gentamicin of use standard MIC, 1.5 doubly to ceftriaxone, Ciprofloxacin, clindamycin and the Ofloxacine USP 23 of standard MIC, 2.0 doubly to the erythromycin of standard MIC, the suis quick medicine-sensitive test-results of carrying out with the EN nutrient solution is identical with the scraps of paper method of diffusion of standard.
Table 1 EN nutrient solution selects to cultivate the effect of clinical milk sample streptococcus intermedius
| Milk sample number | EN liquid culture (suis) | Blood agar plate (suis) | Baird-Parker flat board (staphylococcus | Maconkey agar flat board (intestinal bacteria) |
| 2740 | + | + | - | - |
| 1004 | + | + | - | - |
| 7169 | + | + | + | - |
| 1202 | + | + | - | - |
| 2621 | + | + | - | - |
| 8009 | + | + | + | - |
| 9114 | + | + | + | + |
| 6125 | + | + | + | - |
| 8072 | + | + | + | - |
| 0041 | + | + | - | + |
| 7015 | + | + | - | - |
| 2724 | + | + | + | + |
| 0065 | + | + | + | - |
| 2661 | + | + | - | - |
| Do not have number | + | + | + | + |
| 2668 | + | + | - | - |
| 1148 | + | + | + | + |
| 2789 | + | + | + | + |
"+" and "-" represents respectively and do not have a bacterial growth.
Table 2 international standard (NCCLS20012) suppresses the explanation and relevant minimal inhibitory concentration of circle about the streptococcus susceptibility
The digitized representation antibacterial circle diameter; R represents resistance, and I represents medium sensitivity, and S represents responsive.
Two kinds of streptococcus drug sensitive test methods of table 3 result relatively
| Microbiotic | The suis numbering | Paper disk method (mm) | The EN liquid culture method | |||
| 0.5MIC | 1.0MIC | 1.5MIC | 2.0MIC | |||
| Penicillin | 99082LF | S(40) | - | - | - | - |
| 1033RF | S(24) | - | - | - | - | |
| 0048RH | S(40) | - | - | - | - | |
| 2020LF | R(19) | + | + | + | + | |
| Ceftriaxone | 0048RH | S(29) | - | - | - | - |
| 1033RF | S(28) | - | - | - | - | |
| 9930RH | S(29) | + | + | - | - | |
| 1408 | R(15) | + | + | + | + | |
| Ciprofloxacin | 99082LF | S(19) | + | - | - | - |
| 9938 | S(23) | + | - | - | - | |
| 1408 | S(24) | + | - | - | - | |
| 9830RF | R(12) | + | + | + | + | |
| Clindamycin | 99082LF | S(220 | - | - | - | - |
| 0048RH | S(21) | + | + | - | - | |
| 1033RH | S(24) | - | - | - | - | |
| 9830 | R(13) | + | + | + | + | |
| Erythromycin | 1408 | S(31) | - | - | - | - |
| 9830RF | S(23) | + | + | - | - | |
| 2019LF | S(22) | + | + | + | - | |
| 0033 | I(18) | + | + | + | + | |
| Tsiklomitsin | 9830 | S(24) | - | - | - | - |
| 9938 | S(26) | - | - | - | - | |
| 2024 | S(23) | - | - | - | - | |
| 0033 | R(13) | + | + | + | + | |
| Ofloxacine USP 23 | 99082LF | S(19) | + | - | - | - |
| 0033 | S(19) | + | - | - | - | |
| 2020LF | S(16) | + | + | - | - | |
| 9830 | S(22) | - | - | - | - | |
| Gentamicin | 99082LF | S(19) | - | - | - | - |
| 0033 | S(20) | - | - | - | - | |
| 9830 | S(21) | - | - | - | - | |
| 9938 | R(7) | - | - | - | - | |
Digitized representation antibacterial circle diameter in the bracket; R represents resistance, and I represents medium sensitivity, and S represents responsive; MIC is the minimal inhibitory concentration of international standard in the table.
Table 4 carries out the required suitable antibiotic concentration of quick medicine-sensitive test with the EN liquid culture method
| Microbiotic | Penicillin | Ceftriaxone | Ciprofloxacin | Clindamycin | Erythromycin | Tsiklomitsin | Ofloxacine USP 23 | Gentamicin * |
| MIC | 1.0X | 1.5X | 1.5X | 1.5X | 2.0X | 1.0X | 1.5X | 1.0X |
| Actual concentrations (μ g/ml) | 0.12 | 0.75 | 3 | 0.375 | 0.5 | 2 | 3 | 500 |
*: the bag of gentamicin is to be used for the screening of streptococcus faecium high level aminoglycoside resistance by concentration; MIC in the table refers to the minimal inhibitory concentration of international standard.
EN liquid culture method and standard disk diffusion method drug sensitive test result that table 5 carries out with suis positive milk sample and pure culture
| Strain number | Method | Microbiotic | |||||||
| Penicillin | Ceftriaxone | Ciprofloxacin | Clindamycin | Erythromycin | Tsiklomitsin | Ofloxacine USP 23 | Gentamicin * | ||
| 9830 | Paper disk method | S | S | S | R | S | S | S | S |
| The EN/ pure culture | - | - | - | + | - | - | - | - | |
| The EN/ sample of suckling | - | - | - | + | - | - | - | - | |
| 9938 | Paper disk method | S | S | S | R | S | S | S | R |
| The EN/ pure culture | - | - | - | + | - | - | - | - | |
| The EN/ sample of suckling | - | - | - | + | - | - | - | - | |
| 1079 | Paper disk method | S | S | S | R | R | S | S | S |
| The EN/ pure culture | - | - | - | + | + | - | - | - | |
| The EN/ sample of suckling | - | - | - | + | + | - | - | - | |
| 0033 | Paper disk method | S | S | R | R | R | R | S | R |
| The EN/ pure culture | - | - | + | + | + | + | - | - | |
| The EN/ sample of suckling | - | - | + | + | + | + | - | - | |
| 9946 | Paper disk method | S | S | S | S | S | S | S | R |
| The EN/ pure culture | - | - | - | - | - | - | - | - | |
| The EN/ sample of suckling | - | - | - | - | - | - | - | - | |
| 1408 | Paper disk method | S | S | S | R | S | S | S | S |
| The EN/ pure culture | - | - | - | + | - | - | - | - | |
| The EN/ sample of suckling | - | - | - | + | - | - | - | - | |
| 2020LF | Paper disk method | R | R | R | R | S | R | S | S |
| The EN/ pure culture | + | + | + | + | - | + | - | - | |
| The EN/ sample of suckling | + | + | + | + | - | + | - | - | |
| 2019LF | Paper disk method | S | S | R | R | S | S | S | R |
| The EN/ pure culture | - | - | + | + | - | - | - | - | |
| The EN/ sample of suckling | - | - | + | + | - | - | - | - | |
S represents responsive; R represents resistance; + and-represent respectively in proofreading and correct MIC and EN nutrient solution, grow (resistance) and do not grow (sensitivity).
Claims (3)
1, a kind of compound method of cow mammitis streptococcus culture fluid, it is characterized in that: 10g peptone, 5g yeast powder, 10g sodium-chlor, 5g glucose and 0.5g sodium azide are dissolved in the 1L water, adjust pH to 7.2 with sodium hydroxide solution, add aseptic 0.1% Viola crystallina aqueous solution 1.3ml and 1% purpurum bromocresolis aqueous solution 1.5ml again, 121 ℃ of sterilizations down, standby in 4 ℃ of preservations at last.
2, streptococcus culture fluid according to claim 1 is used to differentiate the method for cow mammitis streptococcus, it is characterized in that: inoculation milk sample in containing the cultivation plate hole of nutrient solution, 37 ℃ down cultivate 24 hours after, nutrient solution becomes yellow by purple and is judged as in the milk sample suis is arranged.
3, streptococcus culture fluid according to claim 1 is used for the drug sensitive test of cow mammitis streptococcus, it is characterized in that: nutrient solution is sub-packed in is coated with antibiotic culture plate, the inoculation suis is selected culture or the positive milk of mastitis sample for the first time, cultivated 24 hours down at 35 ℃, nutrient solution becomes yellow by purple and is judged as microbiotic is had susceptibility.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2008131897A (en) * | 2006-11-28 | 2008-06-12 | Nissui Pharm Co Ltd | Culture medium for detecting enterococcus |
| CN104593286A (en) * | 2014-11-27 | 2015-05-06 | 苏州嘉禧萝生物科技有限公司 | Streptococcus agalactiae culture medium and preparation method thereof |
| CN106350571A (en) * | 2016-08-30 | 2017-01-25 | 北京市农林科学院 | Method for rapidly screening sensitive antibiotic for treating cow mastitis |
| CN107532194A (en) * | 2015-02-27 | 2018-01-02 | 马斯塔普莱克斯有限公司 | Discrimination of bacteria and antimicrobial sensitivity tests |
| CN116334170A (en) * | 2023-05-26 | 2023-06-27 | 南京农业大学三亚研究院 | Identification medium for streptococcus agalactiae of bovine origin |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1099271A (en) * | 1993-08-23 | 1995-03-01 | 王世荣 | Ecological oral liquid |
| CN1672539A (en) * | 2005-04-04 | 2005-09-28 | 陕西科技大学 | Production process of sour milk type lactobacillus milk powder |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
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| JP2008131897A (en) * | 2006-11-28 | 2008-06-12 | Nissui Pharm Co Ltd | Culture medium for detecting enterococcus |
| CN104593286A (en) * | 2014-11-27 | 2015-05-06 | 苏州嘉禧萝生物科技有限公司 | Streptococcus agalactiae culture medium and preparation method thereof |
| CN107532194A (en) * | 2015-02-27 | 2018-01-02 | 马斯塔普莱克斯有限公司 | Discrimination of bacteria and antimicrobial sensitivity tests |
| CN106350571A (en) * | 2016-08-30 | 2017-01-25 | 北京市农林科学院 | Method for rapidly screening sensitive antibiotic for treating cow mastitis |
| CN106350571B (en) * | 2016-08-30 | 2019-09-24 | 北京市农林科学院 | A kind of method of the sensitive antibiotics of quick screening treatment mastitis for milk cows |
| CN116334170A (en) * | 2023-05-26 | 2023-06-27 | 南京农业大学三亚研究院 | Identification medium for streptococcus agalactiae of bovine origin |
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