CN101642475A - Microbial preparation for preventing and treating mastitis and applications thereof - Google Patents
Microbial preparation for preventing and treating mastitis and applications thereof Download PDFInfo
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- CN101642475A CN101642475A CN200910042263A CN200910042263A CN101642475A CN 101642475 A CN101642475 A CN 101642475A CN 200910042263 A CN200910042263 A CN 200910042263A CN 200910042263 A CN200910042263 A CN 200910042263A CN 101642475 A CN101642475 A CN 101642475A
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Abstract
The invention discloses a microbial preparation for preventing and treating mastitis and applications thereof. Bdellovibrio telotroch and Bdellovibrio plastid are prepared into the microbial preparation for preventing and treating mastitis. By detection, the microbial preparation has favorable functions for preventing and treating mastitis of pigs, sheep and cows. The microbial preparation using the mixture of the Bdellovibrio telotroch and the Bdellovibrio plastid for controlling and eliminating mastitis pathogen has the advantages of quick effect taking and strong activity of the telotroch and the advantages of easy preparation and storage, long sterilization action time and strong environmental resistance. Since Bdellovibrio does not have toxic or side effect on cows, the microbial preparation prepared from the Bdellovibrio telotroch and the Bdellovibrio plastid is suitable for large-size factory culture of pigs, sheep and cows, and can provide a novel method for green processing and production of meat products and milk products and for preventing and treating mastitis pathogen.
Description
Technical field
The present invention relates to a kind of microorganism formulation, particularly a kind of microorganism formulation and application thereof that prevents and treats mastitis.
Background technology
Mastitis (also being mastitis) are a kind of diseases that is common, complicated, that cause milch cow, sheep, the serious economic loss of pig aquaculture.The survey showed that on ground such as China Beijing, Shanghai milk cow recessive mastitis sickness rate is about 60%.At present the whole world has 2.2 hundred million cow heads approximately, wherein has 1/3 milch cow to suffer from various types of mastitis approximately, and the annual loss that causes because of mastitis is up to 35,000,000,000 dollars.Although this disease has been carried out 100 years of researches, fail to propose a way that thoroughly solves so far.
Pathogenic infection is the main cause that causes mastitis, and antibacterial, fungus, virus, mycoplasma etc. are arranged.Wherein main pathogenic bacterium are antibacterials, and common have staphylococcus aureus, escherichia coli, Streptococcus and a mycoplasma.Staphylococcus and streptococcus are the The main pathogenic fungi of clinical type mastitis, account for more than 50%, and secondly be escherichia coli.Mastitis due to these three kinds of pathogen take place throughout the year, and the mastitis due to other pathogen mostly are and distribute or secondary.
At present the method for conventional control mastitis mainly contains and does the breast phase pour into antibiotic in milch cow, sheep, pig mammary gland, and antibiotic can be selected optimal drug according to the popular pathogenic bacteria in locality, directly medicine is injected in the breast, the drug level height, absorb fast, good effect.But shortcoming is to produce zest to mammary gland tissue, can cause new inflammation.If long-term a large amount of antibiotic that uses can make the pathogenic bacterium Resistant strain increase, some mastitis cases become and are difficult to cure with conventional method.In addition, antibiotic remains can influence milk quality in the milk.Therefore market does not allow to sell the excessive milk of antibiotic remains, causes Milk Production person's economic loss simultaneously yet.
The herbal control mastitis have drug residue free, advantage that economic benefit is high, but the slow shortcoming that takes effect is arranged.What recombinant vaccine adopted the vaccine control mastitis is safer, effective method.But utilizing the modal problem of vaccine control bovine mastitis is the variation of bacterial antigens, causes vaccine fine 1 year effect, and 1 year ineffective.Other some therapies comprise, mammary gland massage, regimen and homeopathy etc., and their major advantage is to be free from side effects and the residue problem that can not cause the milk Chinese medicine, pathogenic bacterium can not develop immunity to drugs.But, all consuming time, the consumption power of major part in these therapies, also need sick cattle individual processing.The modern breeding scale of their incompatibilities, production efficiency is lower.
Bdellovibrio is to parasitize other antibacterials, and can cause the cracked bacterioid of host bacteria.Littler than general antibacterial, can pass through bacterial filter, the effect of similar phage is arranged.Unicellular, curved or funny point-like, shape in the shape of a spiral sometimes, size (0.2～0.6) * (0.8～1.2) micron, end is given birth to flagellum, mostly is one.Gram is negative, can the cracking gram negative bacteria.Our current research shows that some Bdellovibrio can cracking comprise some common bovine mastitis pathogenic bacterium such as escherichia coli, staphylococcus aureus, streptococcus dysgalactiae.The life cycle of Bdellovibrio divides two stages: free living, can move telotroch stage of not breeding and the trematodiasis plastid stage of in the periplasmic space of specific host antibacterial, carrying out growth and breeding.The Bdellovibrio telotroch is meant the growth forms before Bdellovibrio is invaded host bacteria.Bdellovibrio is in the telotroch stage, and identification also with the host irreversible fixation takes place, and invades the host then, and it is very fast that the Bdellovibrio telotroch is invaded the process of host bacteria, can finish in general several seconds.Bdellovibrio trematodiasis plastid is Bdellovibrio carries out growth and breeding in the periplasmic space of specific host antibacterial a form.After Bdellovibrio entered host's bacterial cell periplasmic space, mainly the macromole by the degraded host cell became monomer, took in then to be used for the synthetic of its biomacromolecule.Along with cell proliferation and hydrolytic enzyme produce, make host cell wall film destroyed, thereby cracking discharge Bdellovibrio.
As control with eliminate the microorganism formulation of mastitis pathogenic bacterium, it is rapid-action to have telotroch concurrently with the mixture of Bdellovibrio telotroch and trematodiasis plastid, energetic and trematodiasis plastid preparation, preserves easily the more of a specified duration and stronger advantage of environment tolerance of bactericidal action time.Make the Bdellovibrio mixture be suitable as and restrain or remove the biological cleaning factor of pathogenic bacterium in organism and the environment thereof, thereby overcome side effect that the antibiotic abuse brings and routine disinfection agent adverse effect milch cow and milk quality.
Summary of the invention
Primary and foremost purpose of the present invention is to overcome the microorganism formulation weak point of not using Bdellovibrio trematodiasis plastid and telotroch preparation control mastitis in the prior art as yet simultaneously, and a kind of microorganism formulation of preventing and treating mastitis is provided.
Another object of the present invention is to provide the application of described microorganism formulation.
Purpose of the present invention is achieved through the following technical solutions: a kind of microorganism formulation of preventing and treating mastitis is made of Bdellovibrio telotroch and Bdellovibrio trematodiasis plastid;
Described Bdellovibrio trematodiasis plastid is that Bdellovibrio is present in the state in the host bacterium;
Described Bdellovibrio telotroch is the state that Bdellovibrio is free in host bacterium outside;
Described Bdellovibrio trematodiasis plastid is preparation by the following method preferably: at the DNB that contains the host bacterium (dilutenutrient broth) fluid medium (nutrient broth 0.8g, caseinic acid hydrolysate 0.5g, yeast extract 0.1g, be dissolved in the 1000ml distilled water, pH value is 7.2～7.6) middle access Bdellovibrio, in 25～35 ℃, 150～300rpm cultivates 36～48h, obtain containing the culture fluid of Bdellovibrio trematodiasis plastid and Bdellovibrio telotroch, culture fluid is behind 4 ℃ of centrifugal 15～20min of 6000～8000rpm, removal contains the supernatant of Bdellovibrio telotroch, and the precipitation that obtains promptly is a Bdellovibrio trematodiasis plastid;
Described host bacterium is a mastitis pathogenic bacterium;
Described mastitis pathogenic bacterium is more elected the bovine mastitis pathogenic bacterium as;
Described bovine mastitis pathogenic bacterium comprise escherichia coli (Escherichiacoli), staphylococcus aureus (Staphyloccocus aureus), staphylococcus epidermidis (Staphylococcusepidermidis), streptococcus agalactiae (Streptococcus agalactia) and the streptococcus dysgalactiae (Streptococcusdysgalactiae) that can cause bovine mastitis;
Described Bdellovibrio trematodiasis plastid is preferably used DNB fluid medium (nutrient broth 0.8g, caseinic acid hydrolysate 0.5g, yeast extract 0.1g is dissolved in the 1000ml distilled water, and pH value is 7.2～7.6), water, normal saline or 0.2mol/L pH value be that 7.2～7.6 phosphate buffer suspends;
The preparation method of described Bdellovibrio telotroch for the supernatant that contains the Bdellovibrio telotroch in the Bdellovibrio trematodiasis plastid preparation method behind the centrifugal 15～20min of 16000～18000rpm, the precipitation that obtains promptly is the Bdellovibrio telotroch;
Described Bdellovibrio telotroch is preferably used DNB fluid medium (nutrient broth 0.8g, caseinic acid hydrolysate 0.5g, yeast extract 0.1g is dissolved in the 1000ml distilled water, and pH value is 7.2～7.6), water, normal saline or 0.2mol/L pH value be that 7.2～7.6 phosphate buffer suspends;
Described Bdellovibrio is preferably one or both of Bdellovibrio (Bdellovibrio sp.) BDS01 or Bdellovibrio (Bdellovibriosp.) BDS02;
Preferred Bdellovibrio telotroch of described microorganism formulation and Bdellovibrio trematodiasis plastid are pressed bacterial population and are mixed at 1: 1;
Described Bdellovibrio (Bdellovibrio sp.) BDS01 is preserved in Chinese typical culture collection center on August 7th, 2009, and deposit number is CCTCC NO:M 209169; Morphologic observation under ultramicroscope: BDS01 was unicellular after described Bdellovibrio BDS01 carried out negative staining, arc, and size is 0.9 * 0.25 μ m, and end is given birth to flagellum, and flagellum length is 4 μ m; Described Bdellovibrio BDS01 cultivates the transparent circular plaque that can form diameter 1～2mm in four days with the double-layer plate method in 28 ℃;
Described Bdellovibrio (Bdellovibrio sp.) BDS02 is preserved in Chinese typical culture collection center on August 7th, 2009, and deposit number is CCTCC NO:M 209170; Morphologic observation under ultramicroscope: BDS02 was unicellular after described Bdellovibrio BDS02 carried out negative staining, arc, and size is 0.65 * 0.2 μ m, end is given birth to flagellum, flagellum length at least 1.8 μ m; Described Bdellovibrio BDS02 cultivates the transparent circular plaque that can form diameter 1～2mm in four days with the double-layer plate method in 28 ℃;
Described microorganism formulation is applied to prevent and treat mastitis;
Described mastitis are preferably bovine mastitis, Lac caprae seu ovis adenitis or pig mastitis;
When described microorganism formulation was used to prevent mammiferous mastitis, mammal was healthy mammal, was to handle mammiferous mammary gland with the soaking flushing or the method for spraying, if soaking flushing, the time is 1miin; If spray, cow mammary gland is evenly sprayed twice, accomplish not stay the dead angle; Bdellovibrio concentration wherein reaches 10 at least
3Pfu/ml, its preferable range is 10
5～10
8Pfu/ml;
When described microorganism formulation was used for the treatment of mammiferous mastitis, mammary gland soaked 1min for having infected the mammary gland of mastitis at every turn, if spray, evenly sprayed twice at every turn; Every day 2 times, continue 5 days; The concentration of used Bdellovibrio reaches 10 at least
3Pfu/ml, its preferable range is 10
5～10
8Pfu/ml.
The present invention has following advantage and effect with respect to prior art:
1, the effect of microorganism formulation of the present invention in the mammiferous mastitis of control is remarkable.
Shown in table 3～5, the microorganism formulation of preventing and treating the mammalian milk adenitis described in the present invention has remarkable result in the treatment to the mammalian milk adenitis, and the therapeutic effect with common antibiotics is suitable basically.Microorganism formulation is to healthy mammal prevention mastitis effect also highly significant.
2, microorganism formulation of the present invention safety in preventing and treating the application of mammalian milk adenitis is good.
In the microorganism formulation Bdellovibrio to eliminate the method that can cause the bovine mastitis pathogenic bacterium by cracking are biological methods.Bdellovibrio can be infected, the characteristic of cracking host bacteria makes it to be suitable as the biological cleaning factor of restraining or removing pathogenic bacterium in organism and the environment thereof, and it is behind the intact host bacteria of cracking, can wither away automatically because of hungry, thereby overcome the side effect that the antibiotic abuse brings.
3, contain control microorganism formulation of the present invention and be fit to be applied to mammal breeding process such as milch cow, sheep, pig.
Bdellovibrio has no side effect to mammals such as people and milch cow, sheep, pigs.Be suitable for large-scale industrialized breed such as milch cow, sheep, pig, can provide safeguard for the green processing production of meat products and milk product and provide a kind of new method for preventing and treating the mammalian milk adenitis.
5, it is applied widely to contain the microorganism formulation of control bovine mastitis of the present invention, also applicable to the people.
The specific embodiment
The present invention is described in further detail below in conjunction with embodiment, but embodiments of the present invention are not limited thereto.
Embodiment 1
(1) preparation of the microorganism formulation of control mastitis
The Bdellovibrio bacteriovorus bacterial strain that uses is respectively Bdellovibrio BDS01 and Bdellovibrio BDS02.Bdellovibrio BDS01 is preserved in Chinese typical culture collection center on August 7th, 2009, and deposit number is CCTCC NO:M 209169; Morphologic observation under ultramicroscope: BDS01 was unicellular after described Bdellovibrio BDS01 carried out negative staining, arc, size are 0.9 * 0.25 μ m, and end is given birth to flagellum, flagellum length is 4 μ m, and it is cultivated the transparent circular plaque that can form diameter 1～2mm in four days with the double-layer plate method in 28 ℃.Described Bdellovibrio BDS02 is preserved in Chinese typical culture collection center on August 7th, 2009, and deposit number is CCTCCNO:M 209170; Morphologic observation under ultramicroscope: BDS02 was unicellular after described Bdellovibrio BDS02 carried out negative staining, arc, size is 0.65 * 0.2 μ m, end is given birth to flagellum, flagellum length at least 1.8 μ m cultivate the transparent circular plaque that four day can form diameter 1～2mm with the double-layer plate method in 28 ℃ with it.
The preparation of Bdellovibrio trematodiasis plastid: at 2 100mL LB (Luria-Bertani) fluid medium (peptone 10g is housed respectively, yeast extract 5g, distilled water 1000mL, pH 7.2) conical flask in inoculate escherichia coli (available from microorganism fungus kind preservation center, Guangdong Province, numbering GIM 1.137), 200rpm, 28 ℃ of shaking tables were cultivated 18 hours, and culture fluid is abandoned supernatant respectively at 4 ℃, the centrifugal 15min of 5000rpm.Respectively precipitation is joined 2 100mL DNB fluid medium (nutrient broth 0.8g is housed, caseinic acid hydrolysate 0.5g, yeast extract 0.1g, be dissolved in the 1000ml distilled water, pH value is 7.2～7.6) conical flask in, add the plaque of the Bdellovibrio BDS01 of picking from the double-layer plate and the plaque of Bdellovibrio BDS02 more respectively.Constant temperature shaking table 250rpm, 28 ℃ of cultivation 36h.Culture fluid is removed the supernatant that contains the Bdellovibrio telotroch respectively at 4 ℃ of centrifugal 20min of 6000rpm, keeps precipitation (being Bdellovibrio trematodiasis plastid), adds normal saline suspension Bdellovibrio trematodiasis plastid;
The preparation of Bdellovibrio telotroch: with the supernatant that contains the Bdellovibrio telotroch in the Bdellovibrio trematodiasis plastid preparation method behind the centrifugal 20min of 16000rpm, the precipitation that obtains promptly is the Bdellovibrio telotroch, add DNB fluid medium (nutrient broth 0.8g, caseinic acid hydrolysate 0.5g, yeast extract 0.1g, be dissolved in the 1000ml distilled water, pH value is 7.2～7.6) suspension Bdellovibrio telotroch.
Microorganism formulation A is 10 by concentration
3Bdellovibrio BDS01 telotroch bacterium liquid and the concentration of pfu/ml are 10
3The Bdellovibrio BDS01 trematodiasis plastid bacterium liquid of pfu/ml is pressed bacterial population and is mixed composition at 1: 1;
Microorganism formulation B is 10 by concentration
3Bdellovibrio BDS02 telotroch bacterium liquid and the concentration of pfu/ml are 10
3The Bdellovibrio BDS02 trematodiasis plastid bacterium liquid of pfu/ml is pressed bacterial population and is mixed composition at 1: 1;
Microorganism formulation A and microorganism formulation B are pressed bacterial population mixing in 2: 1, obtain microorganism formulation C;
Microorganism formulation A and microorganism formulation B are pressed bacterial population mixing in 1: 1, obtain microorganism formulation D;
Microorganism formulation A and microorganism formulation B are pressed bacterial population mixing in 1: 2, obtain microorganism formulation E.
(2) the sick milch cow of mastitis is infected in the microorganism formulation treatment
A, trial drug: experimental group: the microorganism formulation of step (1) preparation
Matched group: ciprofloxacin lactate (Luoyang Huizhong Veterinary Medicine Co. Ltd.'s production);
B, treatment case: treat clinical type mastitis 138 examples altogether, wherein ciprofloxacin lactate group and microorganism formulation A, B, C, D, each 23 example of E group;
C, dosage and method: dosage is as follows: ciprofloxacin lactate 1.5%20mL; Bdellovibrio 10
3Pfu/ml1000mL; Administrated method is as follows: ciprofloxacin lactate is the injection of breast district, with percent by volume 75% cotton ball soaked in alcohol sterilization nipple, make the full and uniform diffusion of medicinal liquid with hands massage cream district 3～5min after having annotated medicinal liquid, use 1% iodine tincture dipping nipple 1min at last, every day 1 time, logotype 5 days; Bdellovibrio is for soaking mammary gland 1min or evenly spraying mammary gland twice, every day 2 times, logotype 5 days;
D, curative effect determinate standard: divide clinical recovery, effective and invalid three kinds; Clinical recovery: the breast red and swollen heat pain disappears firmly, and the milk perusal is normal, and milk yield gos up, and detects the Lac Bovis seu Bubali somatic number less than 500,000/mL; Effectively: the breast red and swollen heat pain is clearly better firmly, and milk does not have grumeleuse or a small amount of milk piece is only arranged, milk yield go up or recovery from illness drug withdrawal 5 days in recidivist again, detect the Lac Bovis seu Bubali somatic number between 1,000,000/mL and 2,000,000/mL; Invalid: the hard nothing of breast red and swollen heat pain is obviously improved or is more serious, and milk is unusual, and milk yield continues to descend, and detection Lac Bovis seu Bubali somatic number is greater than 2,000,000/mL.
The detection of Lac Bovis seu Bubali somatic number adopts international dairy industry community (IDF) to recommend the somatic cell counting method that adopts.The concrete regulation of Lac Bovis seu Bubali somatic number recommended hour grade standard sees Table 1.Concrete test process: at first use each nipple end of percent by volume 75% alcohol wash, and discard 1-2, get the 2-5mL breast that each nipple squeezes and put into corresponding little plate milk.In each little plate, add detection by quantitative reagent, can judge according to reacting phenomenon and California, USA breast method of testing (CMT method) criterion.The criterion of CMT method sees Table 2.
The result is as shown in table 3, and the therapeutic effect of microorganism formulation A group and microorganism formulation B group still also has certain therapeutic effect to bovine mastitis not as good as the antibiotic group.The therapeutic effect of microorganism formulation C, D, E group is suitable with the antibiotic therapy group.And microorganism formulation D group therapeutic effect is best.Bdellovibrio is eliminated pathogen on the mammary gland surface by cracking, can not exert an influence to milk cow health, and the long-term treatment pathogenic bacterium can not develop immunity to drugs, and also can not cause antibiotic residual in milk.May obtain significant more effect if strengthen the Bdellovibrio consumption.Bdellovibrio treatment bovine mastitis operation is simple, easy and simple to handle, and cost is also low.Bdellovibrio has very big value and application potential in the treatment of bovine mastitis.
Table 1, Lac Bovis seu Bubali somatic number grade scale
Table 2, the criterion of CMT method
Annotate: divide in 1 week of puerperium or the cow of late lactation all strong positive reaction can occur, promptly the somatic cell number average physiological occurs and raises in colostrum phase and the lactogenic late lactation milk, so this method is unsuitable for checking the newborn sample in colostrum phase and lactogenic latter stage.
The sick cattle effect of bovine mastitis is infected in the treatment of table 3 microorganism formulation
Embodiment 2
The cold sheep diseases sheep of the little tail of mastitis is infected in the microorganism formulation treatment, and detailed process is as follows:
The microorganism formulation of A, trial drug: experimental group: embodiment 1 step (1) preparation
Matched group: ciprofloxacin lactate (Luoyang Huizhong Veterinary Medicine Co. Ltd.'s production);
B, treatment case: treat clinical type mastitis 120 examples altogether, wherein ciprofloxacin lactate group and microorganism formulation A, B, C, D, each 20 example of E group;
C, dosage and method: dosage is as follows: ciprofloxacin lactate 1.5%10mL; Bdellovibrio 103pfu/ml1000mL; Administrated method is as follows: 1. squeeze clean milk earlier, with percent by volume 75% alcohol disinfecting nipple, milk duct is inserted mammary gland, inject ciprofloxacin lactate, catch nipple to massage breast from bottom to top in turn then, make medicinal liquid be spread in whole mammary gland, every day 2 times, logotype 5 days.2. 50g/L iodine tincture 100mL embrocates skin of breast (noting not being applied to pars papillaris); Bdellovibrio is for soaking mammary gland 1min or evenly spraying mammary gland twice, every day 2 times, logotype 5 days;
D, curative effect determinate standard: divide clinical recovery, effective and invalid three kinds; Clinical recovery: breast redness, heating, pain disappear, and breast does not have lump, and the milk perusal is normal, and milk yield gos up; Effectively: breast redness, heating, pain are clearly better, and milk does not have grumeleuse or a small amount of milk piece is only arranged, milk yield go up or recovery from illness drug withdrawal 5 days in recidivist again; Invalid: breast redness, heating, palpation performance pain, lactogenic reduces or stops the milk degeneration.
The result is as shown in table 4, and the therapeutic effect of microorganism formulation A group and microorganism formulation B group is poor slightly, and the therapeutic effect that microorganism formulation C, E organize is more remarkable, microorganism formulation D with antibiotic therapy group therapeutic equivalence.On the whole, microorganism formulation is lower than milch cow to the treatment cure rate of milk sheep, but effective percentage is higher than milch cow.
Mastitis disease sheep effect is infected in the treatment of table 4 microorganism formulation
Embodiment 3
(1) preparation of the microorganism formulation of control sow mastitis
Preparation process is with embodiment 1 step (1), and the solution of used suspension trematodiasis plastid and telotroch was with PBS buffer (0.2mol/L, pH value are 7.2) when difference only was to prepare microorganism formulation A, B, C, D, E.
(2) the mastitis disease pig is infected in the microorganism formulation treatment
A, trial drug: experimental group: the microorganism formulation of step (1) preparation
Matched group: 0.5-1% procaine hydrochloride, penicillin (Dikang Science and Technology Pharmaceutical Co., Ltd., Sichuan Prov's production);
B, treatment case: treat clinical type mastitis 120 examples altogether, wherein ciprofloxacin lactate group and microorganism formulation A, B, C, D, each 20 example of E group;
C, dosage and method: dosage is as follows: procaine hydrochloride 0.5-1%15mL, penicillin 40 million international units; Bdellovibrio 10
3Pfu/ml 1000mL administrated method is as follows: squeeze clean milk earlier, YUSHIZHI RUANGAO is coated in the part, adds penicillin 40 million international units with 15 milliliters of 0.5-1% procaine hydrochlorides and does breast district closed injection, every day 1 time, logotype 5 days; Bdellovibrio is for soaking mammary gland 1min or evenly spraying mammary gland twice, every day 2 times, logotype 5 days;
D, curative effect determinate standard: divide clinical recovery, effective and invalid three kinds; Clinical recovery: breast flash, swelling, generate heat, harden, pain disappears, the milk perusal is normal; Effectively: breast flash, swelling, generate heat, harden, pain is clearly better, milk do not have grumeleuse a small amount of milk piece is only arranged or recovery from illness drug withdrawal 5 days in recidivist again; Invalid: as breast flash, swelling, to generate heat, harden, have pain, do not allow the piglet sucking.Initial stage milk is thin, interior mixed cotton-shaped fritter.Back hypogalactia and dense is mixed with white floccule, is with the blood streak sometimes even the yellowish-brown pus is arranged, frowziness.
Mastitis disease pig effect is infected in the treatment of table 5 microorganism formulation
The result is as shown in table 5, and the therapeutic effect of microorganism formulation A group and microorganism formulation B group is remarkable not as the therapeutic effect of microorganism formulation C, E group, and microorganism formulation D therapeutic effect is best.Though therapeutic effect not as antibiotic, still can not work the mischief to the healthy of sow and piglet, life-time service can not develop immunity to drugs yet, and the concentration of increasing microorganism formulation has significant more effect.
The foregoing description is a preferred implementation of the present invention; but embodiments of the present invention are not restricted to the described embodiments; other any do not deviate from change, the modification done under spirit of the present invention and the principle, substitutes, combination, simplify; all should be the substitute mode of equivalence, be included within protection scope of the present invention.
Claims (7)
1, a kind of microorganism formulation of preventing and treating mastitis is characterized in that: described microorganism formulation is made of Bdellovibrio telotroch and Bdellovibrio trematodiasis plastid.
2, microorganism formulation according to claim 1 is characterized in that: described Bdellovibrio is that preservation date is that August 7, deposit number in 2009 Bdellovibrio (Bdellovibrio sp.) BDS01 that is CCTCC NO:M 209169 or preservation date are one or both of August 7, deposit number in 2009 Bdellovibrio (Bdellovibrio sp.) BDS02 that is CCTCC NO:M 209170.
3, microorganism formulation according to claim 1 is characterized in that: described microorganism formulation is that Bdellovibrio telotroch and Bdellovibrio trematodiasis plastid mix by bacterial population at 1: 1.
4, according to each described microorganism formulation of claim 1～3, it is characterized in that: described Bdellovibrio trematodiasis plastid prepares by the following method: containing DNB fluid medium (the nutrient broth 0.8g of host bacterium, caseinic acid hydrolysate 0.5g, yeast extract 0.1g, be dissolved in the 1000ml distilled water, pH value is 7.2～7.6) middle access Bdellovibrio, cultivate 36～48h in 25～35 ℃, 150～300 rpm, culture fluid is behind 4 ℃ of centrifugal 15～20min of 6000～8000rpm, removal contains the supernatant of Bdellovibrio telotroch, and the precipitation that obtains promptly is a Bdellovibrio trematodiasis plastid;
Behind the centrifugal 15～20min of 16000～18000rpm, the precipitation that obtains promptly is described Bdellovibrio telotroch with the described supernatant that contains the Bdellovibrio telotroch.
5, the application of each described microorganism formulation of claim 1～3 is characterized in that: described microorganism formulation is applied to prevent and treat mastitis.
6, application according to claim 5 is characterized in that: described mastitis are bovine mastitis, Lac caprae seu ovis adenitis or pig mastitis.
7, application according to claim 5 is characterized in that: described microorganism formulation is handled mammary gland, and the concentration of the Bdellovibrio in the described microorganism formulation reaches 10 at least
3Pfu/ml.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101999456A (en) * | 2010-08-31 | 2011-04-06 | 华南理工大学 | Method for controlling total number of bacteria in fresh milk |
| CN102007940A (en) * | 2010-08-31 | 2011-04-13 | 华南理工大学 | Application of bdellovibrio mixture in preparing turbot red bill disease preventing and curing bactericide |
| CN102038714A (en) * | 2010-08-31 | 2011-05-04 | 华南理工大学 | Application of bdellovibrio bdelloplast in preparing bacterial agent for preventing and controlling red mouth disease of scophthalmus maximus |
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2009
- 2009-08-28 CN CN200910042263A patent/CN101642475A/en active Pending
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101999456A (en) * | 2010-08-31 | 2011-04-06 | 华南理工大学 | Method for controlling total number of bacteria in fresh milk |
| CN102007940A (en) * | 2010-08-31 | 2011-04-13 | 华南理工大学 | Application of bdellovibrio mixture in preparing turbot red bill disease preventing and curing bactericide |
| CN102038714A (en) * | 2010-08-31 | 2011-05-04 | 华南理工大学 | Application of bdellovibrio bdelloplast in preparing bacterial agent for preventing and controlling red mouth disease of scophthalmus maximus |
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Open date: 20100210 |