CN1724998A - Diagnosis reagent for dairy cattle recessive mammitis and its preparation method - Google Patents
Diagnosis reagent for dairy cattle recessive mammitis and its preparation method Download PDFInfo
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- CN1724998A CN1724998A CN 200510049389 CN200510049389A CN1724998A CN 1724998 A CN1724998 A CN 1724998A CN 200510049389 CN200510049389 CN 200510049389 CN 200510049389 A CN200510049389 A CN 200510049389A CN 1724998 A CN1724998 A CN 1724998A
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- reagent
- milk
- mammitis
- powder
- cmt
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- 208000004396 mastitis Diseases 0.000 title claims abstract description 29
- 239000003153 chemical reaction reagent Substances 0.000 title claims description 38
- 241000283690 Bos taurus Species 0.000 title claims description 20
- 238000003745 diagnosis Methods 0.000 title claims description 16
- 235000013365 dairy product Nutrition 0.000 title claims description 8
- 238000002360 preparation method Methods 0.000 title claims description 6
- 239000000843 powder Substances 0.000 claims abstract description 18
- KGBXLFKZBHKPEV-UHFFFAOYSA-N boric acid Chemical compound OB(O)O KGBXLFKZBHKPEV-UHFFFAOYSA-N 0.000 claims abstract description 8
- 239000004327 boric acid Substances 0.000 claims abstract description 8
- 235000010339 sodium tetraborate Nutrition 0.000 claims abstract description 8
- 239000007787 solid Substances 0.000 claims abstract description 7
- 229910021538 borax Inorganic materials 0.000 claims abstract description 6
- 239000004328 sodium tetraborate Substances 0.000 claims abstract description 6
- 239000000203 mixture Substances 0.000 claims description 8
- JZVZLMSUWQYJRJ-UHFFFAOYSA-N C1(=CC=CC=C1)O.[K].[Br] Chemical compound C1(=CC=CC=C1)O.[K].[Br] JZVZLMSUWQYJRJ-UHFFFAOYSA-N 0.000 claims description 7
- RCEAADKTGXTDOA-UHFFFAOYSA-N OS(O)(=O)=O.CCCCCCCCCCCC[Na] Chemical compound OS(O)(=O)=O.CCCCCCCCCCCC[Na] RCEAADKTGXTDOA-UHFFFAOYSA-N 0.000 claims description 7
- 238000000034 method Methods 0.000 abstract description 9
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 abstract description 7
- 238000001514 detection method Methods 0.000 abstract description 5
- 239000007788 liquid Substances 0.000 abstract description 4
- 239000003795 chemical substances by application Substances 0.000 abstract 2
- 241000245063 Primula Species 0.000 abstract 1
- 235000016311 Primula vulgaris Nutrition 0.000 abstract 1
- DBMJMQXJHONAFJ-UHFFFAOYSA-M Sodium laurylsulphate Chemical compound [Na+].CCCCCCCCCCCCOS([O-])(=O)=O DBMJMQXJHONAFJ-UHFFFAOYSA-M 0.000 abstract 1
- ABIUHPWEYMSGSR-UHFFFAOYSA-N bromocresol purple Chemical compound BrC1=C(O)C(C)=CC(C2(C3=CC=CC=C3S(=O)(=O)O2)C=2C=C(Br)C(O)=C(C)C=2)=C1 ABIUHPWEYMSGSR-UHFFFAOYSA-N 0.000 abstract 1
- 229940039227 diagnostic agent Drugs 0.000 abstract 1
- 239000000032 diagnostic agent Substances 0.000 abstract 1
- 239000008267 milk Substances 0.000 description 31
- 235000013336 milk Nutrition 0.000 description 31
- 210000004080 milk Anatomy 0.000 description 31
- 210000004027 cell Anatomy 0.000 description 15
- 230000000392 somatic effect Effects 0.000 description 14
- 238000012360 testing method Methods 0.000 description 13
- 239000004033 plastic Substances 0.000 description 6
- 229920003023 plastic Polymers 0.000 description 6
- 210000000481 breast Anatomy 0.000 description 5
- 238000006243 chemical reaction Methods 0.000 description 5
- 239000000047 product Substances 0.000 description 4
- 230000000052 comparative effect Effects 0.000 description 2
- 239000012141 concentrate Substances 0.000 description 2
- 238000002405 diagnostic procedure Methods 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 239000012153 distilled water Substances 0.000 description 2
- 238000000105 evaporative light scattering detection Methods 0.000 description 2
- 238000010438 heat treatment Methods 0.000 description 2
- BASFCYQUMIYNBI-UHFFFAOYSA-N platinum Chemical compound [Pt] BASFCYQUMIYNBI-UHFFFAOYSA-N 0.000 description 2
- 238000001556 precipitation Methods 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- 238000010998 test method Methods 0.000 description 2
- 235000002710 Ilex cornuta Nutrition 0.000 description 1
- 241001310146 Ilex cornuta Species 0.000 description 1
- 235000010326 Osmanthus heterophyllus Nutrition 0.000 description 1
- 239000000853 adhesive Substances 0.000 description 1
- 230000001070 adhesive effect Effects 0.000 description 1
- 230000004520 agglutination Effects 0.000 description 1
- 210000000170 cell membrane Anatomy 0.000 description 1
- 238000000546 chi-square test Methods 0.000 description 1
- 239000000470 constituent Substances 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 230000008034 disappearance Effects 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 239000008151 electrolyte solution Substances 0.000 description 1
- JEIPFZHSYJVQDO-UHFFFAOYSA-N ferric oxide Chemical compound O=[Fe]O[Fe]=O JEIPFZHSYJVQDO-UHFFFAOYSA-N 0.000 description 1
- 210000004969 inflammatory cell Anatomy 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 238000000386 microscopy Methods 0.000 description 1
- 229910052697 platinum Inorganic materials 0.000 description 1
- 239000002244 precipitate Substances 0.000 description 1
- 239000013049 sediment Substances 0.000 description 1
- 210000001082 somatic cell Anatomy 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
Abstract
The invention discloses a cow recessive mastitis diagnostic agent and the preparing method, uniformly mixing borax 50-70 shares, boric acid 47-49 shares, and bromcresol purple 1 share, then mixing with dodecyl sodium sulfate 190-210 shares, thus making primrose solid powder, taking the powder 15.45 g to dissolve in 500 ml water so as to form purplish-red liquid, able to be used for diagnosing the cow recessive mastitis. The agent is the same as CMT and can indirectly determine somatocyte content of a cow, simple to operate and able to make field detection, and especially, the powder of the agent has more advantages, such as small bulk, lightweight, convenience of carrying and storing and extremely suitable for large- volume detection.
Description
Technical field
The present invention relates to a kind of diagnosis reagent for dairy cattle recessive mammitis and preparation method thereof.
Technical background
The stealthy mammitis of milk cow is ignored by people easily because of there not being macroscopic clinical symptoms.But the incidence of disease is higher, and is easy to change into clinic mastitis.According to the U.S., the milk cow of general 48-49% suffers from stealthy mammitis.Domestic because management level and sanitary condition are relatively poor, and situation is even more serious, and the incidence of disease can reach more than the 65-70%, suffer from milk crop and quality and significantly reduce, so its harmfulness is especially big.
In the milk of recessive mastitis ox, the time except constituent and the change of physicochemical character physicochemical property.Somatic number also rolls up.The diagnostic method that uses is checked its somatic number more at present.Directly microscopy method workload is big and time-consuming.Milk body cell automatic counter for counting cost height.Therefore, adopt indirect method to increase several scopes to understand its body cell more.Indirect method is from the application thirties, through improving, founded California, USA mammitis test method(s) (CMT) to the later stage fifties.Various countries adopt CMT more or create the method that improves certainly later on.Domestic research mastitis for milk cows is started late.There is report the beginning eighties, as the Lanzhou mammitis diagnosticum (LMT) of Lanzhou Chinese veterinarian's Research Institute.The SMT in the BMT of Pekinese, Shanghai etc.But above all kinds of solution or the concentrate of being, transportation and storage are inconvenient.
Summary of the invention
The purpose of this invention is to provide a kind of diagnosis reagent for dairy cattle recessive mammitis and preparation method thereof.
Diagnosis reagent for dairy cattle recessive mammitis contains 190-210 part lauryl sodium sulfate, 50-70 part borax, 47-49 part boric acid and 1 part of bromine potassium phenol violet.
The preparation method of diagnosis reagent for dairy cattle recessive mammitis mixes 50-70 part borax, 47-49 part boric acid and 1 part of bromine potassium phenol violet, mix with 190-210 part lauryl sodium sulfate again, the faint yellow solid powder.
The identical body cell content that can indirect determination milk of reagent of the present invention with CMT, easy and simple to handle, can detect at the scene, particularly its pulvis has more advantage, and is little as volume, in light weight, be easy to carry and storage etc., extremely be fit to carry out the usefulness of mass detection.
Embodiment
Embodiment 1
50 parts of boraxs, 47 parts of boric acid and 1 part of bromine potassium phenol violet are mixed, mix with 190 parts of lauryl sodium sulfate again, the faint yellow solid powder.This powder is divided in the plastic bottle, and every bottle contains powder 15.45 grams, standby.
Before using the powder in 1 bottle is dissolved among the warm water, final volume is 500 milliliters, and pH is 7.3～7.5, and this liquid can be used for the diagnosis of cow subclinical mastitis.
Embodiment 2
70 parts of boraxs, 49 parts of boric acid and 1 part of bromine potassium phenol violet are mixed, mix with 210 parts of lauryl sodium sulfate again, the faint yellow solid powder.This powder is divided in the plastic bottle, and every bottle contains powder 15.45 grams, standby.
Before using the powder in 1 bottle is dissolved among the warm water, final volume is 500 milliliters, and pH is 7.3～7.5, and this liquid can be used for the diagnosis of cow subclinical mastitis.
Embodiment 3
With boric acid (Guanghua Science ﹠ Technology Co., Ltd., Shanghai's product) 240 grams, borax (granary, Jiangsu chemical industry two factory's products) 300 grams and bromine potassium phenol violet (reagent company limited product is thought carefully in Shanghai) 5 grams mix, again and lauryl sodium sulfate (Xiamen star grand reach (Sanland) chemical reagent company limited product) 1000 grams mix, the faint yellow solid powder.This powder is divided in the plastic bottle, and every bottle contains powder 15.45 grams, standby.
Before using the powder in 1 bottle is dissolved among the warm water, final volume is 500 milliliters, and pH is 7.3～7.5, and this liquid can be used for the diagnosis of cow subclinical mastitis.
Embodiment 4
Three kinds of cow subclinical mastitis diagnostic methods comparison
One, materials and methods
(1) reagent and instrument
1, mastitis for milk cows diagnosis solid powder of the present invention faces the solution that becomes 500 milliliters with dissolved in distilled water with preceding.
2, California, USA mammitis diagnosticum (CMT) is 1 pint of original-pack concentrate, according to directions for use, uses by dilution in 1: 7 with distilled water.
3, mammitis plastics diagnosis dish.
4, quantitative sample injector.
5, Coulter cell counter, the ZM type, Britain Coulter company makes.Use 100 microns aperture pipes, regulate instrument according to the instrument instructions.Once Ji Shuo sample size is 100 microlitres.
6, Somafix reagent and Somaton reagent, Coulter company produces.
7, the attached plastics suction nozzle of the quantitative pipettor of 100 microlitres.
8,100 ℃ and 65 ℃ of water baths are each 1.
(2) test method
1, milk sample collection: the milk sample picks up from this school and Hangzhou five happinesses, Dinghai, 4 cattle farms of Chinese holly.15 milliliters of foremilks are got in test tube by each breast district, add a cover rubber stopper after, take back the laboratory and check immediately.
2, reagent of the present invention and CMT check: respectively add 2 milliliters of milk samples in two disks of mammitis plastics diagnosis dish, add 2 milliliters of reagent of the present invention and CMT reagent more respectively, shake the diagnosis dish then, observe and judge level of reaction.
Level of reaction criterion:
Negative (-): evenly mix, any precipitation do not occur.
Suspicious (T): micro-fines then disappear after shaking.
Weak positive (+): a small amount of viscous precipitate, mobile big when shaking, not agglutination.
Positive (++): precipitation is many, is gluey.Sediment has adhering sense when shaking, and assembles to the center,
Strong positive (+++): be gluey rapidly, assemble to the center when shaking, be central protuberance, toughness is big, at the bottom of the adhesive disc.
3, milk somatic cell counting: undertaken by the instrument instructions; The concrete operations step is as follows: (1) fixed body cell membrane: at first add 4 Somafix reagent in test tube, add 10 milliliters of milk samples again, put will suckle on the motorized agitator sample and the even test tube of reagent mix and add a cover foraminate rubber stopper; Place 60 ℃ of water-bath heating 5 minutes then, it is cooled to room temperature voluntarily to take out relief.(2) destroy fat globule: in clean test tube, add 10 milliliters of Somaton reagent earlier, add milk sample 100 microlitres of fixed cell film with quantitative sample injector again.After test tube is added a cover the rubber stopper that has aperture, be placed on heating taking-up in 10 minutes in 80 ℃ of water-baths, treat to be cooled to voluntarily room temperature.(3) body cell counting: will pour in 10 milliliters the small beaker through the sample of above-mentioned processing, and be put on the Coulter counter and measure, every duplicate samples detection twice, the result gets its mean value.
Every milliliter of somatic number=200 * (instrument readings-blank counts)
Two, result
(1) relation of reagent reacting grade of the present invention and somatic number.Use reagent of the present invention and Coulter body cell counter and detected 797 parts of milk samples simultaneously.Through significance test, very significantly (P＜0.001=illustrates that the level of reaction of reagent of the present invention raises with the increase of Ruzhong somatic number purpose to gradient.The body cell content of each grade milk sample of reagent of the present invention sees Table 1.
The body cell content of each grade milk sample of table 1 reagent of the present invention
| Reagent reacting grade of the present invention | Check breast district number | Every ml milk somatic number mean value | Standard deviation (SD) |
| - | 188 | 78000 | 48000 |
| T | 119 | 277000 | 98000 |
| + | 245 | 758000 | 281000 |
| ++ | 99 | 1930000 | 844000 |
| +++ | 146 | 6590000 | 524000 |
(2) reagent of the present invention and CMT contrast test.It the results are shown in Table 2.By Chi-square Test, the routine number of each level of reaction of two kinds of method mensuration does not have significant difference (P＞0.05)
Table 2 reagent of the present invention and CMT comparative test result
| Level of reaction | - | T | + | ++ | +++ | Add up to * |
| HMT checks breast district number | 188 | 119 | 245 | 99 | 146 | 797 |
| CMT checks breast district number | 183 | 116 | 242 | 100 | 139 | 780 |
*, 17 parts of milk sample factor amounts are arranged very little, do not make CMT and check.
Three, discuss
When (one) milk cow suffered from mammitis, a large amount of inflammatory cells was assembled to ill breast district, thereby the somatic number in Ruzhong is increased, and its quantity what can reflect that seriously whether ill degree.This test is that the more advanced accurate Coulter cell counter of application is measured the body cell number in the milk sample.The testing process and the principle of this instrument are: milk is through handling, and body cell promptly is suspended in the electrolyte solution; After the sample of handling put into counter, through the both sides of a special this aperture in aperture family a platinum electrode is housed respectively, has Huan to flow electric current between two electrodes and pass through even be loaded on an amount of sample flow of the quantitative sample injection parts of instrument internal.Somatic resistance is bigger, when it during through small holes, will form pulse voltage, the counting circuit of instrument is had an effect, make its work, count, and the result shown.Coulter cell counter good reproducibility, the accuracy height is widely used in some developed countries at present.
(2) as can be seen by result listed in the table 1, reagent of the present invention is determined as-, T ,+, ++, ++ it is 78,000,277,000,758,000,1,930 for average respectively every milliliter that+milk sample at different levels contains somatic number, 000 and 6,590,000.By the standard that Schalm etc. is formulated, CMT every milliliter of somatic numbers that contain at different levels should be 0-2000, and 000,150,040-500,000,400,000-1,500,000,800,000-5,000,000 and＞5,000,000.This shows that this test is used every milliliter of contained somatic mean of each grade milk sample of reagent mensuration of the present invention all within the due somatic number scope of CMT respective level.Numerical value in the table 2 shows, the umber basically identical of each grade sample that two kinds of methods of reagent of the present invention and CMT determine equated as comparative test result with former institute.Therefore can prove that the detection performance of HMT pulvis is identical with CMT, the reagent as quick diagnosis milk cow disappearance mammitis can replace the latter to use fully.
(3) according to the regulation of international milk cow federation, the somatic number in every milliliter in the milk surpasses 500,000, is mammitis and suffers from milk, should be handled or not edible.Some developed countries act in accordance with it to this regulation.In order to protect health, China must formulate corresponding regulation by lattice, and strictness is carried out.The identical body cell content that can indirect determination milk of reagent of the present invention with CMT, easy and simple to handle, can detect at the scene, particularly its pulvis has more advantage, and is little as volume, in light weight, be easy to carry and storage etc., extremely be fit to carry out the usefulness of mass detection.
Four, conclusion
The milk sample grade that reagent of the present invention is measured raises with the increase of Ruzhong somatic number.The body cell content and the basically identical of using the original-pack CMT reagent detection of the U.S. of each grade milk sample that reagent of the present invention is measured.Therefore, reagent of the present invention fully can replaced C MT as the reagent of quick test mammitis.
Claims (2)
1. a diagnosis reagent for dairy cattle recessive mammitis is characterized in that, it contains 190-210 part lauryl sodium sulfate, 50-70 part borax, 47-49 part boric acid and 1 part of bromine potassium phenol violet.
2. the preparation method of a diagnosis reagent for dairy cattle recessive mammitis is characterized in that, 50-70 part borax, 47-49 part boric acid and 1 part of bromine potassium phenol violet are mixed, mix with 190-210 part lauryl sodium sulfate again, the faint yellow solid powder.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB200510049389XA CN100483114C (en) | 2005-03-17 | 2005-03-17 | Diagnosis reagent for dairy cattle recessive mammitis and its preparation method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB200510049389XA CN100483114C (en) | 2005-03-17 | 2005-03-17 | Diagnosis reagent for dairy cattle recessive mammitis and its preparation method |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1724998A true CN1724998A (en) | 2006-01-25 |
| CN100483114C CN100483114C (en) | 2009-04-29 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB200510049389XA Expired - Fee Related CN100483114C (en) | 2005-03-17 | 2005-03-17 | Diagnosis reagent for dairy cattle recessive mammitis and its preparation method |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101825626A (en) * | 2010-04-22 | 2010-09-08 | 中国农业科学院兰州畜牧与兽药研究所 | Diagnostic reagent for detecting cow subclinical mastitis and preparation method thereof |
| CN101865922A (en) * | 2010-05-17 | 2010-10-20 | 北京易斯威特生物医学科技有限公司 | Milk somatic cell detection kit and method |
| CN102213715A (en) * | 2010-04-08 | 2011-10-12 | 内蒙古硕高生物科技有限责任公司 | New animal mastitis detection method and detector |
| CN106841636A (en) * | 2017-01-24 | 2017-06-13 | 北京美正生物科技有限公司 | A kind of enzyme-linked immunosorbent assay kit for detecting cow subclinical mastitis and its production and use |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101942522B (en) * | 2010-10-14 | 2012-07-04 | 扬州大学 | Assay kit for identifying dairy cow mastitis resistance-related molecular marker |
-
2005
- 2005-03-17 CN CNB200510049389XA patent/CN100483114C/en not_active Expired - Fee Related
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102213715A (en) * | 2010-04-08 | 2011-10-12 | 内蒙古硕高生物科技有限责任公司 | New animal mastitis detection method and detector |
| CN101825626A (en) * | 2010-04-22 | 2010-09-08 | 中国农业科学院兰州畜牧与兽药研究所 | Diagnostic reagent for detecting cow subclinical mastitis and preparation method thereof |
| CN101865922A (en) * | 2010-05-17 | 2010-10-20 | 北京易斯威特生物医学科技有限公司 | Milk somatic cell detection kit and method |
| CN106841636A (en) * | 2017-01-24 | 2017-06-13 | 北京美正生物科技有限公司 | A kind of enzyme-linked immunosorbent assay kit for detecting cow subclinical mastitis and its production and use |
Also Published As
| Publication number | Publication date |
|---|---|
| CN100483114C (en) | 2009-04-29 |
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Effective date of registration: 20160704 Address after: 1 B12A11, 39 building, 310012 Yue Yue Road, Hangzhou, Zhejiang, Xihu District Patentee after: Hangzhou vision Wang Biological Technology Co.,Ltd. Address before: 310027 Hangzhou, Zhejiang Province, Zhejiang Road, No. 38 Patentee before: Zhejiang University |
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Granted publication date: 20090429 |