CN101991611A - Active biological antibacterial and production method thereof - Google Patents
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Abstract
The invention relates to active biological antibacterial and a production method thereof. The active biological antibacterial is prepared by proportionally mixing the cultures of bdellovibrio and bacteriophage serving as the main ingredients in the following steps: obtaining the bdellovibrio culture and the bacteriophage culture respectively, and preparing the cultures at a proportion of 1:1-1:1,000,000, namely that the bdellovibrio content is 10-1,010/pfu, and the bacteriophage content is 10-1,016/pfu; and mixing the cultures so that the cultures are suspended in medium liquid or mixed in excipient. The invention is used for preventing and controlling the bacterial diseases for human body, animals and plants, and has obvious effects on guaranteeing quality and freshness of fruits, vegetables, flowers, food and aquatic products, eliminating harmful bacteria in the environment and suppressing growth of blue-green algae.
Description
Technical field
The present invention relates to a kind of control human body, animal, bacteriosis, really, vegetables, flowers, food, aquatic products guarantee the quality fresh-keeping and remove noxious bacteria in the environment, suppress biological preparation---the active bio antibacterial and the production method thereof of blue algae growth, belong to biological product and manufacturing field thereof.
Background technology
Be used at present the control medicine of bacterial disease both at home and abroad, mainly contain antibiotic and chemosynthesis antibacterials.The use of this type of medicine, the one, constantly produce drug resistance strain, effectively dosage increases gradually, causes the generation to all antibacterials all insensitive " superbacteria "; The 2nd, residual in vivo, suppress the metabolism of body normal growth, bring disaster to health; The 3rd, contaminated environment can be detrimental to health equally.Therefore, method and the goods of developing the Biological control bacterial disease of high effect nontoxic have caused great interest of people and concern, and the research of all kinds of biological and ecological preparations emerges, and extremely payes attention to the biological control method of bacterium system bacterium.
The Bdellovibrio that find the sixties is the parasitics antibacterial that a class is made a living with predator bacteria specially, has the unique biological characteristic of " parasitism " and " dissolving (killing) " noxious bacteria, and it is littler than general antibacterial usually.The effect of Bdellovibrio can selectively be adsorbed onto on host's bacterial cell and enter in host's bacterial cell, and growth and breeding finally causes the cracking of host's bacterial cell therein.It all has parasitic and cracked effect to the noxious bacteria in many human bodies, animal, plant pathogen and the environment, and this unique biological characteristic has caused the great attention of researcher.But for a long time, both at home and abroad researcher thinks that always the growth temperature of Bdellovibrio is 20-30 ℃, and optimum growth temperature is 26.3 ℃, just can not growth and breeding more than 35 ℃, and can only on viable bacteria, grow.And the intravital normal temperature of people is generally about 37 ℃, and the intravital normal temperature of animal is generally about 40 ℃.Except that Bdellovibrio to temperature sensitivity especially, the Bdellovibrio of cultivating in the environment is easy inactivation in vivo; On viable bacteria, cultivate again in the production process of Bdellovibrio, the easy contaminated environment of viable bacteria, and cultivation cycle is long, and harvest yield is low, and viable bacteria is residual harmful to environment in the culture; Particularly outstanding is, Bdellovibrio is wide to host's cracking scope, sensitivity is strong, but specificity is not as good as phage.
Therefore people utilize Bdellovibrio control human body, animal, bacteriosis, really, vegetables, flowers, food, aquatic products guarantee the quality fresh-keeping and remove noxious bacteria in the environment, the development and application that suppresses blue algae growth is greatly limited.
Phage is the bacterial virus that a class of 20 beginnings of the century discovery can be dissolved (killing) antibacterial.Phage has specificity highly to host's bacterial cell, and the host range of phage is a useful characteristic of strain identification.The treatment that phage is used for some bacterial disease has unique effect, the especially disease that Resistant strain is caused.Phage is to be adsorbed onto on the specific acceptor site of host's bacterial cell with caudal sheath, shrink head and in host's bacterial cell body, inject DNA by tailpin, and in host's bacterial cell body replicon generation, the mode of final similar Bdellovibrio dissolving host bacterial cell discharges filial generation.Phage can not be in stopping metabolic host's bacterial cell growth and breeding.Phage is to the too late Bdellovibrio of the sensitivity of host bacterium, but phage is to corresponding host bacterium high special.
Bdellovibrio culture and phage culture are united use, make the sensitivity of Bdellovibrio and the high degree of specificity of phage have complementary advantages, and significant synergism is arranged.Can eliminate fully and use antibiotic and chemical synthetic drug develops immunity to drugs, residual in the body, the drawback that suppresses normal growth metabolism, contaminated environment.
Summary of the invention
The objective of the invention is to be to provide a kind of mixed culture with Bdellovibrio and phage is main component, be used to prevent and treat human body, animal, bacteriosis, really, vegetables, flowers, food, aquatic products guarantee the quality fresh-keeping and remove noxious bacteria in the environment, suppress biological preparation---the active bio antibacterial and the production method thereof of blue algae growth.
The technical solution that realizes the object of the invention is: a kind of active bio antibacterial is that culture with Bdellovibrio and phage is that Main Ingredients and Appearance is mixed in proportion and makes, and it is characterized in that the mixed culture of Bdellovibrio and phage prepares by the following method:
(1) acquisition of Bdellovibrio culture: the Bdellovibrio host bacterium that makes inactivation earlier, be the host with this inactivation host bacterium then, produce the Bdellovibrio culture with the Bdellovibrio general culture method, select on the double-deck agar plate of tap water plaque typical again, energy cracking noxious bacteria, do not destroy beneficial bacteria, the Bdellovibrio bacteriovorus bacterial strain that in 10-43 ℃ of environment, can grow, then inactivation host bacterium that makes in the said process and the Bdellovibrio that selects are suspended in the diluent, proofread and correct its pH value 3.8-10.0, fermentation culture in 10-43 ℃ of environment, fermentation culture medium or with standby after the fermentation culture medium lyophilizing.
(2) acquisition of phage culture
Produce responsive phage host bacteria suspension, produce the phage culture with the double-deck agar culture method of phage, select the virulent phage of the typical energy of plaque cracking noxious bacteria on the double-deck agar plate of nutrient broth again, single plaque that the picking transparency is good is soaked in the nutrient broth, then, get supernatant and make the double-deck agar plate of nutrient broth again, at last phage culture and the responsive host bacteria suspension that makes in the said process is suspended in the nutrient broth, proofread and correct its pH value 3.8-10.0, fermentation culture in 10-43 ℃ of environment, fermentation culture medium is removed responsive host bacterium through aseptic filtration, determines aseptic fermentation culture medium or the fermentation culture medium lyophilizing is standby through checking.
(3) with above-mentioned Bdellovibrio fermentation culture medium or with fermentation culture medium filtering and concentrating lyophilized powder and through check determine aseptic phage fermentation culture medium or with the fermentation culture medium lyophilized powder in 1: 1-1: 1000000 ratio preparation, promptly the content of Bdellovibrio is 10-10
10Pfu/ml, the content of phage are 10-10
16Pfu/ml; Mixed preparing makes it be suspended in medium liquid or is blended in the excipient; Promptly obtain product.
Above-mentioned steps (1) specifically can be: produce host bacteria suspension with conventional method; Heating then, or, make the inactivation host bacteria suspension with chloroform (chloroform) processing; Be the host with this inactivation host bacteria suspension then, produce its culture with the Bdellovibrio general culture method, select on the double-deck agar plate of tap water plaque typical again, the corresponding noxious bacteria of energy cracking, do not destroy the Bdellovibrio bacteriovorus bacterial strain of beneficial bacteria, then, make double-deck agar plate again, heightening temperature cultivates, select the Bdellovibrio bacteriovorus bacterial strain that in 10-43 ℃ of environment, can grow, at last with the inactivation host bacteria suspension that makes in the said process and the Bdellovibrio that selects in 10: 1-105: 1 ratio is suspended in the diluent, proofreading and correct its pH value is 3.8-10.0, fermentation culture in 30-43 ℃ of environment, fermentation culture medium or with standby after the fermentation culture medium lyophilizing.
Above-mentioned steps (2) concrete operations are: produce responsive (corresponding noxious bacteria) host bacteria suspension with conventional method, produce its culture with the double-deck agar culture method of phage, select typical, the virulent phage that can the corresponding noxious bacteria of cracking of on the double-deck agar plate of nutrient broth plaque again, single plaque that the picking transparency is good is soaked in the nutrient broth, then, the phage culture that makes in the said process is pressed 1 with responsive (corresponding noxious bacteria) host bacteria suspension: 10-1: 10
5Ratio be suspended in the nutrient broth liquid, proofreading and correct its pH value is 3.8-10.0, fermentation culture in 10-43 ℃ of environment, fermentation culture medium is removed responsive (corresponding noxious bacteria) host bacterium through aseptic filtration.Determine aseptic fermentation culture medium or the fermentation culture medium lyophilizing is standby through checking.
During with this product control people, animal body bacterial disease, be main path, also can be aided with other medicine-feeding ways with the Orally taken product administration; When being used for the aquatic animal preventing and treating bacterial diseases of young, the product mode in the breeding environment water body of directly splashing also can be aided with oral; When preventing and treating bacteriosis with this product, can the root embedding, or foliage spray; Really, vegetables, flowers, food, aquatic products guarantee the quality fresh-keeping to be main route of administration with foliage spray, also can to soak; When removing in the environment noxious bacteria and blue algae, can directly splash in environment water with this product.
Active bio antibacterial provided by the present invention is through Chinese medicine antibacterial preservation administrative center vibrio phage research department test, to the cleavage rate of human body harmful antibacterials such as human pathogen salmonella typhi, escherichia coli, dysentery bacterium, vibrio cholera, vibrio parahaemolytious, bacillus pyocyaneus, staphylococcus aureus more than 75-95.38%.Agricultural University Of Nanjing, Shandong Agricultural University's test, to animal pathogen swine escherichia coli, Pullorum Disease bacillus, Salmonella, pasteurellosis bacillus, yersinia, streptococcus, pseudomonas, Aeromonas hydrophila, vibrio parahaemolyticus, the cleavage rate of animal and plant noxious bacteria such as vibrio alginolyticus, wound vibrio, Vibrio anguillarum, visible peristalsis visible intestinal peristalsis point-like Aeromonas, Flexibacter columnaris, pseudomonas fluorescens and plant pathogen rice leaf spot bacteria, soft rot of cabbage bacterium, Rhizoma Zingiberis Recens ralstonia solanacearum is more than 92.43%.Agricultural University Of Nanjing, Tianjin agriculture university, Agricultural University Of Jiangxi, the test of Jiangsu Province bureau of animal husbandry, product are used to prevent and treat result's demonstration of intestine of young pigs bacterial disease, and the prevention protective rate is 87.46-99.33%, reduce mortality rate more than 99%.Agricultural product quality inspection center, Jiangsu Province, Nanjing science and technology office agricultural place, scientific and technical department of Shandong Province's Laiwu City bureau of agriculture test result show, the active bio antibacterial is used for fruit-vegetable food such as blackberry guarantees the quality when fresh-keeping, can prolong and guarantee the quality freshness date 3-7 days; Be used for the control of Fructus Capsici, Rhizoma Zingiberis Recens bacterial wilt; protective rate is 79.86-94.12%; to the common pathogenic bacterium of plant, be 93.76% as the cleavage rate of the sick Erwinia of handle rest fungus, pseudomonas, rice leaf spot bacteria, carrot soft rot, citrus ulcer Xanthomonas campestris, soft rot of cabbage bacterium etc.
The method of production active bio antibacterial disclosed by the invention avoided cultivating the environmental pollution that Bdellovibrio is caused on viable bacteria, and cultivation cycle is short, output is high; Particularly make Bdellovibrio, phage to preventing and treating bacterial diseases of young human body, animal, bacteriosis, really, vegetables, flowers, food, aquatic products are guaranteed the quality fresh-keeping and remove noxious bacteria in the environment, suppress blue algae growth etc. becomes a reality.
The specific embodiment
Energy kill harmful antibacterial (being meant implication understood by one of ordinary skill in the art) among the present invention is as the most of pathogenic bacterium in Salmonella, vibrio, Shigella, colon bacillus genus, Yersinia, Pseudomonas aeruginosa genus, staphylococcus, the Streptococcus; Protection beneficial bacteria (being meant what implication understood by one of ordinary skill in the art or state food drug regulatory department expressly ratified) is as lactobacillus casei, Lactobacillus plantarum, streptococcus faecalis, pediococcus acidilactici, bacillus subtilis, Bafillus natt, bacillus acidophilus, Streptococcus lactis (Lister) Lohnis 1909.554., beer yeast, Candida utilis, algae pool rhodopseudomonas, Bacillus cereus, bacteroides fragilis etc.The bacterial strain that concrete experiment screening draws among the following embodiment only for explanation method of the present invention, does not constitute the qualification to protection domain of the present invention.
Said Bdellovibrio and phage among the present invention can be asked for or separation acquisition from environment to culture presevation mechanism.And measure the cracking scope of Bdellovibrio and phage respectively.Selection has the Bdellovibrio bacteriovorus bacterial strain and the phage strain of cracking (killing) effect to the most of noxious bacteria in common Salmonella, vibrio, Shigella, colon bacillus genus, Yersinia, Pseudomonas aeruginosa genus, staphylococcus, the Streptococcus.
Host bacterium of the present invention, at first obtaining the non-pathogenic bacteria Bdellovibrio sensitivity, not toxogenic with conventional method is the host bacterium, as e. coli k12, C600, Erwinia, Lactobacillus plantarum etc., reuse heating or the lethal method of chloroform make host bacterium inactivation.Specific practice is (is example with escherichia coli C600): ask for or separate from environment to culture presevation mechanism and obtain escherichia coli (the escherichia coli C600 of this experiment derives from Institute of Microorganism, Academia Sinica), with its separation of on Nutrient agar, ruling, after the cultivation, do corresponding serum coagulation, get the male single bacterium colony of agglutination, be inoculated in the nutrient broth, after the cultivation, the transferred species agar slant is cultivated, with aseptic distillation water elution lawn, remove the nutrient substance in the culture medium, add sterile purified water again and be mixed with bacteria suspension; Heat sterilization perhaps uses chloroform (chloroform) to handle then, promptly obtains the escherichia coli of inactivation.
Secondly, be the host with the escherichia coli of above-mentioned inactivation, at first screen noxious bacteria cracking scope wider relatively, the Bdellovibrio that can not destroy beneficial bacteria again, obtain its culture with the Bdellovibrio general culture method after, again it is tamed.Specific practice is: ask for or separate from environment from culture presevation mechanism and obtain Bdellovibrio, and the escherichia coli host of itself and inactivation added in the semi-solid top-layer agar of tap water mix, be poured over then on the double-deck agar plate of tap water, it is solidified, then cultivate, choose the typical Bdellovibrio of plaque, picking list plaque is soaked in the sterilization tap water, then, get supernatant and make double-deck agar plate again, heighten temperature and cultivate, and repeatedly for several times, until selecting the Bdellovibrio strain that in 10-43 ℃ of environment, can grow.
Then, be the host with the escherichia coli of inactivation, proofread and correct its pH value 3.8-10.0, through fermentation culture, promptly obtain the Bdellovibrio culture, the Bdellovibrio culture also can be lyophilized into powder.
Obtain phage with the phage separation method to the noxious bacteria sensitivity, specific practice is: ask for or separation acquisition pathogenic bacterium bacterial strain from pathogen or environment to culture presevation mechanism, with its separation of on Nutrient agar, ruling, after the cultivation, do corresponding serum coagulation, get single bacterium colony that agglutination is positive, be inoculated in the nutrient broth and cultivate, obtain the pathogenic bacterium culture.Then, with the pathogenic bacterium culture is the host, the phage strain that screening is strong to the noxious bacteria cracking performance, obtain its culture with the phage general culture method after, select the phage strain that the pathogenic bacterium to correspondence that can grow have stronger cracking ability in 10-43 ℃ of environment.
Then, be the host with the pathogenic bacterium of correspondence, proofread and correct its pH value 3.8-10.0, through fermentation culture, promptly obtain the phage culture, the phage culture also can be lyophilized into powder.
At last with the Bdellovibrio culture (or lyophilized culture) that obtains and the phage culture (or lyophilized culture) that obtains in 1: 1-1: 1000000 ratio preparation, promptly the content of Bdellovibrio is 10-10
10Pfu/ml, the content of phage are 10-10
16Pfu/ml.Mix promptly to obtain being used to prevent and treat human body, animal, bacteriosis, really, vegetables, flowers, food, aquatic products guarantee the quality fresh-keeping and remove noxious bacteria in the environment, suppress biological preparation---the active bio antibacterial of blue algae growth.
Embodiment one: Bdellovibrio, phage mixed culture are to the lytic effect of different genera antibacterial
1, bacterium source: Bdellovibrio, phage are provided by Chinese medicine antibacterial preservation administrative center vibrio phage specialized laboratory; The bacterial isolates of different genera is provided by Nat'l Pharmaceutical ﹠ Biological Products Control Institute.
2, the screening of Bdellovibrio bacteriovorus bacterial strain and phage strain: from the Bdellovibrio bacteriovorus bacterial strain that obtains and phage strain, filter out Bdellovibrio Bd39, Bd40, Bd59, Bd76, Bd81, Bd94, Bd98, Bd329, Bd334 and vibrio phage ∮ V3, ∮ M6, ∮ 1a, ∮ 1b to 24 strain different genera antibacterial sensitivities with conventional method, shigella phage ∮ P22, ∮ 119X, ∮ 17, ∮/F2a, typhoid fever phage ∮ Vi, ∮ S154, Escherichia coli phage ∮ T2, ∮ T7, pyocinophages ∮ PE5, ∮ A, ∮ B.
3, the preparation of Bdellovibrio, phage mixed culture:
The preparation of Bdellovibrio culture: referring to Chinese microbiology and Journal of Immunology 1982,2 (1): 12-15.The Bdellovibrio that obtains and the escherichia coli host of inactivation add in the semi-solid top-layer agar of tap water and mix, be poured over then on the double-deck agar plate of tap water, it is solidified, then cultivated 28 hours for 37 ℃, choose the typical Bdellovibrio of plaque, picking list plaque is soaked in the sterilization tap water, then, get supernatant and make double-deck agar plate again, heightening temperature cultivates, and repeatedly for several times, until selecting Bdellovibrio strain Bd39, Bd40, Bd59, Bd76, Bd81, Bd94, Bd98, Bd329, the Bd334 that in 10-43 ℃ of environment, can grow.
Then, be the host with the escherichia coli of inactivation, proofread and correct its pH value to 3.8-10.0, through fermentation culture, promptly obtain the Bdellovibrio culture.
The preparation of phage culture: referring to Bacteriophages IntersciencePublishers, Inc., New York USA. or " phage " 1975, department Ti east, Chen Tingzuo master translate.Publish phage research department, Jiangsu.With corresponding vibrio, shigella, Bacillus typhi, Escherichia coli, the bacillus pyocyaneus culture is the host, and screening is to corresponding vibrio, shigella, Bacillus typhi, Escherichia coli, the phage strain that the bacillus pyocyaneus cracking performance is strong obtains its culture with the phage general culture method.
The Bdellovibrio culture that obtains at last, the content of Bdellovibrio are 10
3-7Pfu/ml; Phage culture, the content of phage are 10
6-13Pfu/ml.
4, the preparation of different genera bacterial cultures: 24 strain different genera antibacterials separate with nutrient agar panel line respectively, cultivate 18 hours for 37 ℃, select the colonies typical transferred species in the nutrient broth 37 ℃ cultivated 4-6 hour, promptly obtain the different genera bacterial cultures.
Experimental design: the content with Bdellovibrio is 10
3-7Pfu/ml, the content of phage are 10
6-13The pfu/ml mixed culture is the cracking of the strain culture in the different Pseudomonas of 20-70 hundred million cfu/ml to 24 strain content.
Use the double-layer tap water agar flat band method, every flat board is the host with hundred million different genera antibacterials of 10-35, adds 0.5ml Bdellovibrio and phage mixed culture simultaneously, cultivates 48,72 hours each observed and recorded results 1 time in 37 ℃.Establishing with escherichia coli C600 simultaneously is the host, checks that the ability of Bdellovibrio, phage mixed culture formation plaque is a matched group.
5, the results are shown in subordinate list 1.
Table 1. Bdellovibrio, phage mixed culture are to the lytic effect of different genera antibacterial
6, conclusion (of pressure testing):
1), Bdellovibrio, phage mixed culture all have splitting action to the pathogenic bacteria of 22 strain different generas, cleavage rate is 100%;
2), Bdellovibrio, phage mixed culture do not have splitting action to 2 strains microbial ecological preparation production commonly used with bacterial strain.
Embodiment two: the protective effect that Bdellovibrio, phage mixed culture infect Cavia porcellus angle conjunctiva
1, bacterium source: Chinese medicine antibacterial preservation administrative center vibrio phage specialized laboratory provides.
2, the screening of Bdellovibrio bacteriovorus bacterial strain and phage strain: from the Bdellovibrio bacteriovorus bacterial strain that obtains and phage strain, filter out Bdellovibrio Bd98, Bd329 and the phage ∮/F2a responsive to shigella flexneri F2 a 301 (product strain) with conventional method.
3, the preparation of Bdellovibrio, phage mixed culture: the bacterial strain that filters out with step 2 is prepared by the method for embodiment one.
4, the preparation of shigella flexneri culture: shigella flexneri F2 a301 separates with nutrient agar panel line, cultivates 18 hours for 37 ℃, select the colonies typical transferred species in the nutrient broth 37 ℃ cultivated 4-6 hour, promptly obtain the shigella flexneri culture.
Experimental design: with Bdellovibrio content is 10
6Pfu, phage content are 10
8Pfu mixed culture and shigella flexneri content are 2.5 * 10
6The cfu culture splashes in the guinea pig eye, and fixing eyelid 2 minutes.Establish Bdellovibrio, phage mixed culture and shigella flexneri culture matched group simultaneously.Observed once in per 24 hours.Continue 5 days.
5, the results are shown in Table 2.
The protective effect that table 2. Bdellovibrio, phage mixed culture infect Cavia porcellus angle conjunctiva
6, conclusion (of pressure testing):
1), Bdellovibrio, phage mixed culture have no stimulation to Cavia porcellus angle conjunctiva;
2), Bdellovibrio, phage mixed culture are 83.3% to Cavia porcellus angle conjunctiva infection protective rate.
Embodiment three: Bdellovibrio, phage mixed culture are measured the lytic effect of Bacillus typhi
1, bacterium source: Bdellovibrio, phage are provided by Chinese medicine antibacterial preservation administrative center vibrio phage specialized laboratory; Bacillus typhi is provided by Jiangsu Province Center for Disease Control (CDC).
2, the screening of Bdellovibrio bacteriovorus bacterial strain and phage strain: from the Bdellovibrio bacteriovorus bacterial strain that obtains and phage strain, filter out Bdellovibrio Bd59, Bd81, Bd98, Bd125, Bd329, Bd334 and Bacillus typhi phage ∮/A, ∮/D, ∮/E, ∮/K, ∮/M1, ∮/36, ∮/53, ∮/96, ∮/Vi to 206 strain Bacillus typhi sensitivities with conventional method.
3, the preparation of Bdellovibrio, phage mixed culture: the bacterial strain that filters out with step 2 is prepared by the method for embodiment one.The content of Bdellovibrio is 10
6Pfu/ml, the content of phage is 10 for every milliliter
9Pfu/ml.
4, the preparation of Bacillus typhi culture: separate with nutrient agar panel line respectively, cultivated 18 hours for 37 ℃, select the colonies typical transferred species in the nutrient broth 37 ℃ cultivated 4-6 hour, promptly obtain the Bacillus typhi culture.
Experimental design: think 10
6Pfu/ml, the content of phage are 10
9The pfu/ml mixed culture is the cracking of 20-30 hundred million cfu cultures to 206 strain Bacillus typhi content, comprising reference culture 5 strains, and local bacterial strain 201 strains (isolating 199 strains in the human faecal mass, isolating 2 liang of strains in the sewage) in Jiangsu.Comprise in the 206 strain Bacillus typhi that as bacteriophage typing be 133 strains of ∮/A, ∮/D, ∮/E, ∮/K, ∮/M1, ∮/36, ∮/53, ∮/96, ∮/Vi and 184 strains that 20 kinds of antibacterials such as ampicillin are different sensitivitys.
Use the double-layer tap water agar flat band method, every flat board is the host with 2,000,000,000 cfu Bacillus typhi, adds 0.5ml Bdellovibrio and phage mixed culture simultaneously, cultivates 48 hours observed and recorded results in 25 ℃.Establishing escherichia coli C600 simultaneously is matched group, checks that the ability of Bdellovibrio, phage mixed culture formation plaque is.
5, the results are shown in subordinate list 3.
Table 3. Bdellovibrio, phage mixed culture are to the lytic effect of Bacillus typhi
6, conclusion (of pressure testing):
1), Bdellovibrio, phage mixed culture have splitting action to 199 strains in the 206 strain Bacillus typhi, cleavage rate is 96.60%;
2), Bdellovibrio, phage mixed culture have splitting action to 105 strains in the 133 strain different bacteriophages type Bacillus typhi, cleavage rate is 98.10%;
3), Bdellovibrio, phage mixed culture have splitting action to 183 strains in the 184 strain drug resistance Bacillus typhi, cleavage rate is 99.50%.
Embodiment four: Bdellovibrio, phage mixed culture are measured the lytic effect of vibrio
1, bacterium source: Bdellovibrio, phage are provided by Chinese medicine antibacterial preservation administrative center vibrio phage specialized laboratory; Vibrio is provided by Nat'l Pharmaceutical ﹠ Biological Products Control Institute, Jiangsu Province Center for Disease Control (CDC).
2, the screening of Bdellovibrio bacteriovorus bacterial strain and phage strain: from the Bdellovibrio bacteriovorus bacterial strain that obtains and phage strain, filter out Bdellovibrio Bd81, the Bd98 of 69 strain vibrio sensitivities and vibrio phage ∮/1b, ∮/1c, ∮/1d, ∮/1j with conventional method.
3, the preparation of Bdellovibrio, phage mixed culture: the bacterial strain that filters out with step 2 is prepared by the method for embodiment one.The content of Bdellovibrio is 10
4Pfu/ml, the content of phage are 10
6Pfu/ml.
4, the preparation of vibrio culture: 69 strain vibrios separate with nutrient agar panel line respectively, cultivate 18 hours for 37 ℃, select the colonies typical transferred species in the nutrient broth 37 ℃ cultivated 4-6 hour, promptly obtain the vibrio culture.
Experimental design: the content with Bdellovibrio is 10
4Pfu/ml, the content of phage are 10
6The pfu/ml mixed culture is the cracking (comprising vibrio cholera 39 strains, NAG vibrios 10 strains, vibrio parahaemolytious 20 strains) of 20-50 hundred million cfu cultures to 69 strain vibrio content.
Use the double-layer tap water agar flat band method, every flat board is the host with 20-50 hundred million cfu vibrios, adds 0.5ml Bdellovibrio and phage mixed culture simultaneously, in 37 ℃ of cultivations, and 48 hours observed and recorded results.Establishing escherichia coli C600 simultaneously is matched group, checks that Bdellovibrio, phage mixed culture form the ability of plaque.
5, the results are shown in subordinate list 4.
Table 4. Bdellovibrio, phage mixed culture are to the lytic effect of vibrio
6, conclusion (of pressure testing):
1), Bdellovibrio, phage mixed culture have splitting action to 32 strains in the 39 strain vibrio cholera, cleavage rate is 82.05%;
2), Bdellovibrio, phage mixed culture have splitting action to 8 strains in the 10 strain NAG vibrios, cleavage rate is 80.00%;
3), Bdellovibrio, phage mixed culture have splitting action to 18 strains in the 20 strain vibrio parahaemolytious, cleavage rate is 90.00%.
Embodiment five: Bdellovibrio, phage mixed culture are to the scavenging action of shigella flexneri in the water body
1, bacterium source: Chinese medicine antibacterial preservation administrative center vibrio phage specialized laboratory provides.
2, the screening of Bdellovibrio bacteriovorus bacterial strain and phage strain: from the Bdellovibrio bacteriovorus bacterial strain that obtains and phage strain, filter out Bdellovibrio Bd329 and the phage ∮/F2a responsive to shigella flexneri F2 a 301 (product strain) with conventional method
3, the preparation of Bdellovibrio, phage mixed culture: the bacterial strain that filters out with step 2 is prepared by the method for embodiment one.The content of Bdellovibrio is 10
6Pfu/ml, the content of phage are 10
8Pfu/ml.
4, the preparation of shigella flexneri culture: the line of shigella flexneri F2 a 301 usefulness nutrient agar panels separates, and cultivates 18 hours for 37 ℃, selects the colonies typical transferred species in nutrient broth, cultivates 4-6 hour, and promptly obtains the shigella flexneri culture for 37 ℃.
Experimental design: in same water body, the content that adds Bdellovibrio simultaneously is 10
3Pfu/ml, the content of phage are 10
4Pfu/ml mixed culture and shigella flexneri content are the 2500cfu culture.Establish the matched group that does not add Bdellovibrio, phage mixed culture and only add the shigella flexneri culture simultaneously.Per 24 hours water samplings once detect the number of shigella flexneri in the water body.Continue 7 days.
5, the results are shown in Table 5.
Table 5. Bdellovibrio, phage mixed culture are to the scavenging action of shigella flexneri in the water body
6, conclusion (of pressure testing):
Bdellovibrio, phage mixed culture are 99.28-99.97% to the clearance rate of shigella flexneri in the water body, compare with matched group, and learning processing (p＜0.01) by statistics has significant differences.
Embodiment six: Bdellovibrio, phage mixed culture are to the scavenging action of vibrio cholera in the river
1, bacterium source: Chinese medicine antibacterial preservation administrative center vibrio phage specialized laboratory provides.
2, Bdellovibrio bacteriovorus bacterial strain. and the screening of phage strain: from the Bdellovibrio bacteriovorus bacterial strain that obtains and phage strain, filter out Bdellovibrio Bd98, Bd329 and choleraphage ∮/1b to vibrio cholera ElTor18001 sensitivity with conventional method
3, the preparation of Bdellovibrio, phage mixed culture: the bacterial strain that filters out with step 2 is prepared by the method for embodiment one
4, the preparation of vibrio cholera culture: vibrio cholera Fl Tor18001 separates with the nutrient agar panel line, cultivates 18 hours for 37 ℃, selects the colonies typical transferred species in nutrient broth, cultivates 4-6 hour, and promptly obtains the vibrio cholera culture for 37 ℃.
Experimental design: in same river, the content that adds Bdellovibrio simultaneously is 10
2Pfu/ml, the content of phage are 10
4Pfu/ml mixed culture and vibrio cholera content are 10
7Cfu culture/ml.Establish simultaneously and do not add Bdellovibrio, phage mixed culture, only add the matched group of vibrio cholera culture.The timing water sampling detects the number of vibrio cholera in the water body later on.Continue 13 days.
5, the results are shown in Table 6.
Table 6. Bdellovibrio, phage mixed culture are to the scavenging action of vibrio cholera in the river
6, conclusion (of pressure testing):
Test group is compared with matched group, and Bdellovibrio, phage mixed culture have tangible scavenging action to vibrio cholera in the river.
Embodiment seven: Bdellovibrio, phage mixed culture are to the scavenging action of NAG vibrios in the river
1, bacterium source: Chinese medicine antibacterial preservation administrative center vibrio phage specialized laboratory provides.
2, the screening of Bdellovibrio bacteriovorus bacterial strain and phage strain: from the Bdellovibrio bacteriovorus bacterial strain that obtains and phage strain, filter out Bdellovibrio Bd98 and NAG vibrios phage ∮/1i to NAG vibrios 17001 sensitivities with conventional method
3, the preparation of Bdellovibrio, phage mixed culture: the bacterial strain that filters out with step 2 is prepared by the method for embodiment one.
4, the preparation of NAG vibrios culture: NAG vibrios 17001 usefulness nutrient agar panels line separates, and cultivates 18 hours for 37 ℃, select the colonies typical transferred species in the nutrient broth 37 ℃ cultivated 4-6 hour, promptly obtain the NAG vibrios culture.
Experimental design: in same river, the content that adds Bdellovibrio simultaneously is 10
3Pfu/ml, the content of phage are 10
5Pfu/ml mixed culture and NAG vibrios content are 10
6Pfu culture/ml.Establish simultaneously and do not add Bdellovibrio, phage mixed culture, only add the matched group of NAG vibrios culture.The timing water sampling detects the number of NAG vibrios in the water body later on.Continue 48 days.
5, the results are shown in Table 7.
Table 7. Bdellovibrio, phage mixed culture are to the scavenging action of NAG vibrios in the river
6, conclusion (of pressure testing):
Test group and matched group compare, and Bdellovibrio, phage mixed culture have tangible scavenging action to NAG vibrios in the river.
Embodiment eight: Bdellovibrio, phage mixed culture are measured aquatic animal pathogenic bacterium lytic effect
1, bacterium source: Bdellovibrio, phage are provided by Chinese medicine antibacterial preservation administrative center vibrio phage specialized laboratory; The aquatic animal pathogenic bacterium are provided by fresh water fishery research center, Chinese aquatic science institute Wuxi and Shanghai Aquatic Products Univ. 9CN).
2, the screening of Bdellovibrio bacteriovorus bacterial strain and phage strain: from the Bdellovibrio bacteriovorus bacterial strain that obtains and phage strain, filter out Bdellovibrio Bd59, Bd81, Bd98, Bd296, Bd329 and Aeromonas hydrophila ∮ PB31, yersinia ∮ YB1, vibrio parahaemolyticus ∮ VP3a01, phagies such as vibrio alginolyticus ∮ VB131, pseudomonas fluorescens ∮ PS039 to 11 strain aquatic animal pathogenic bacterium sensitivities with conventional method.
3, the preparation of Bdellovibrio, phage mixed culture: the bacterial strain that filters out with step 2 is prepared by the method for embodiment one.
4, the preparation of aquatic animal pathogenic bacterium culture: 11 strain aquatic animal pathogenic bacterium separate with the nutrient agar panel line respectively, cultivated 18 hours for 37 ℃, select the colonies typical transferred species in the nutrient broth 37 ℃ cultivated 4-6 hour, promptly obtain aquatic animal pathogenic bacterium culture.
Experimental design: with Bdellovibrio content is 10
6Pfu/ml, phage content are 10
8The pfu/ml mixed culture is 10 to 11 strain aquatic animal pathogenic bacterium content
6-12The cracking of cfu/ml culture.
Use the double-layer tap water agar flat band method, every flat board is the host with 2,000,000,000 cfu aquatic animal pathogenic bacterium, adds 0.5ml Bdellovibrio and phage mixed culture simultaneously, cultivates 48 hours observed and recorded results in 37 ℃.Establishing escherichia coli C600 simultaneously is matched group, checks that Bdellovibrio, phage mixed culture form the ability of plaque.
5, the results are shown in Table 8.
Table 8. Bdellovibrio, phage mixed culture are to the lytic effect of 11 strain aquatic animal pathogenic bacterium
6, conclusion (of pressure testing):
Bdellovibrio, phage mixed culture all have splitting action to 11 strain aquatic animal pathogenic bacterias, and cleavage rate is 100%.Infer that in view of the above Bdellovibrio, phage mixed culture have bigger theory significance and using value to the control of aquatic animal bacterial disease.
Embodiment nine: Bdellovibrio, phage mixed culture are measured the lytic effect of Aeromonas hydrophila
1, bacterium source: Bdellovibrio, phage are provided by Chinese medicine antibacterial preservation administrative center vibrio phage specialized laboratory; Aeromonas hydrophila is provided by the Jiangsu Prov. Inst. of Microbiology.
2, the screening of Bdellovibrio bacteriovorus bacterial strain and phage strain: from the Bdellovibrio bacteriovorus bacterial strain that obtains and phage strain, filter out Bdellovibrio Bd81, Bd296 and Aeromonas hydrophila phage ∮ PB31 to 9 Hygrophilous monad sensitivities with conventional method.
3, the preparation of Bdellovibrio, phage mixed culture: the bacterial strain that filters out with step 2 is prepared by the method for embodiment one.
4, the preparation of Aeromonas hydrophila culture: 9 Hygrophilous monads separate with nutrient agar panel line respectively, cultivate 18 hours for 37 ℃, select the colonies typical transferred species in the nutrient broth 37 ℃ cultivated 4-6 hour, promptly obtain the Aeromonas hydrophila culture.
Experimental design: with Bdellovibrio content is 10
6Pfu/ml, phage content are 10
9The pfu/ml mixed culture is 10 to 9 Hygrophilous monad content
6-12The cracking of cfu/ml culture.
Use the double-layer tap water agar flat band method, every flat board is the host with 2,000,000,000 cfu Aeromonas hydrophilas, adds 0.5ml Bdellovibrio and phage mixed culture simultaneously, cultivates 48 hours observed and recorded results in 37 ℃.Establish escherichia coli C600 matched group simultaneously, check that the ability of Bdellovibrio, phage mixed culture formation plaque is.
5, the results are shown in Table 9.
Table 9. Bdellovibrio, phage mixed culture are to the lytic effect of Aeromonas hydrophila
6, conclusion (of pressure testing):
Bdellovibrio, phage mixed culture all have splitting action to 9 strain aquatic animal pathogenic bacteria Aeromonas hydrophilas, and cleavage rate is 100%.Infer that in view of the above Bdellovibrio, phage mixed culture have bigger theory significance and using value to the control of fish, shrimp, shellfish aquatic animal bacterial disease.
Embodiment ten: Bdellovibrio, phage mixed culture are measured the animal pathogen lytic effect
1, bacterium source: experiment Salmonella choleraesuls C78-2, chickling Salmonella C79-20, Pullorum Disease Salmonella C79-13, chicken colibacillosis C83861, swine escherichia coli C83907 bacterial strain, provided by China Veterinary Drugs Supervisory Inst., Salmonella enteritidis 50041 is provided by Nat'l Pharmaceutical ﹠ Biological Products Control Institute.Bdellovibrio, phage are provided by Chinese medicine antibacterial preservation administrative center vibrio phage specialized laboratory.
2, the screening of Bdellovibrio bacteriovorus bacterial strain and phage strain: from the Bdellovibrio bacteriovorus bacterial strain that obtains and phage strain, filter out Bdellovibrio Bd81, Bd98, Bd296 and phage ∮ P22, ∮ A, ∮ 1533, ∮ T7, ∮ 17 to 6 strain animal pathogen sensitivities with conventional method.
3, the preparation of Bdellovibrio, phage mixed culture: the bacterial strain that filters out with step 2 is prepared by the method for embodiment one.
4, the preparation of animal pathogen culture: Salmonella choleraesuls, the chickling Salmonella, the Pullorum Disease Salmonella, chicken colibacillosis, swine escherichia coli, Salmonella enteritidis separate with the nutrient agar panel line respectively, cultivate 18 hours for 37 ℃, select the colonies typical transferred species in the nutrient broth 37 ℃ cultivated 4-6 hour, promptly obtain the animal pathogen culture.
Experimental design: with Bdellovibrio content is 10
3Pfu/ml, phage content are 10
6The pfu/ml mixed culture is 10 to 6 strain animal pathogen content
8The cracking of cfu/ml culture.
Use the double-layer tap water agar flat band method, every flat board is the host with 1,000,000,000 cfu animal pathogens, adds 0.5ml Bdellovibrio and phage mixed culture simultaneously, cultivates 48 hours observed and recorded results in 37 ℃.Establishing escherichia coli C600 simultaneously is matched group, checks that Bdellovibrio, phage mixed culture form the ability of plaque.
5, the results are shown in Table 10.
Table 10. Bdellovibrio, phage mixed culture are to the lytic effect of animal pathogen
6, conclusion (of pressure testing):
Bdellovibrio, phage mixed culture all have splitting action to 6 strain animal pathogens, and cleavage rate is 100%.Infer that in view of the above Bdellovibrio, phage mixed culture have bigger using value to the control of animal bacteria disease.
Embodiment 11: Bdellovibrio, phage mixed culture are measured the yellow and white dysentery of piglet therapeutic effect
1, bacterium source: Bdellovibrio, phage are provided by Chinese medicine antibacterial preservation administrative center vibrio phage specialized laboratory.
2, the screening of Bdellovibrio bacteriovorus bacterial strain and phage strain: from the Bdellovibrio bacteriovorus bacterial strain that obtains and phage strain, filter out Bdellovibrio Bd81, Bd98 and swine escherichia coli phage ∮ T101 to the swine escherichia coli sensitivity with conventional method.
3, the preparation of Bdellovibrio, phage mixed culture: the bacterial strain that filters out with step 2 is prepared by the method for embodiment one.
4, experimental design: carry out in experimental farm, Agricultural University Of Nanjing Jiangpu.The interior piglet of 30 ages in days that HUANGBAI(sic) dysentery will take place at random in test is divided into two groups: one group of oral Bdellovibrio content is 10
6Pfu/ml, phage content are 10
10Pfu/ml mixed culture 10ml drug treatment, once a day, for three days on end; Another group is an oral gentamicin 100mg drug treatment, once a day, and for three days on end.Then every day the observed and recorded piglet course of disease situation of change, continuous 10 days.
5, the results are shown in subordinate list 11.
Table 11. Bdellovibrio, phage mixed culture are to the therapeutic effect of yellow and white dysentery of piglet
6, conclusion (of pressure testing):
1), Bdellovibrio, phage mixed culture be to the therapeutic effect that has of yellow and white dysentery of piglet, cure rate reaches 100%;
2), Bdellovibrio, phage mixed culture be better than gentamycin to the therapeutic effect of yellow and white dysentery of piglet, cure rate is higher than gentamycin 29.42%.
Embodiment 12: Bdellovibrio, phage mixed culture are observed the yellow and white dysentery of piglet prevention effect
1, bacterium source: Bdellovibrio, phage are provided by Chinese medicine antibacterial preservation administrative center vibrio phage specialized laboratory.
2, the screening of Bdellovibrio bacteriovorus bacterial strain and phage strain: from the Bdellovibrio bacteriovorus bacterial strain that obtains and phage strain, filter out Bdellovibrio Bd81, Bd98 and swine escherichia coli phage ∮ T101 to Zhu escherichia coli sensitivity with conventional method.
3, the preparation of Bdellovibrio, phage mixed culture: the bacterial strain that filters out with step 2 is prepared by the method for embodiment one.
4, experimental design: carry out in Nanchang City, Jiangxi Province and Zhangshu City.Test is conventional with Bdellovibrio, phage mixed culture prevention HUANGBAI(sic) dysentery with the annual piglet that breeds in pig farm, Nanchang City in 2008 and Zhangshu City pig farm, and the piglet that HUANGBAI(sic) dysentery takes place is treated with Bdellovibrio, phage mixed culture.
Prophylactic tria, adopting the oral cavity to gavage a 5ml Bdellovibrio content same day from the piglet birth is 10
6Pfu/ml, phage content are 10
10The pfu/ml mixed culture was strengthened once in the time of ten days; Another group is an oral ofloxacin 50mg drug treatment, strengthens once in the time of ten days.Then every day the observed and recorded piglet HUANGBAI(sic) dysentery generation incidence.Continuous 30 days.
Therapeutic test, the piglet that HUANGBAI(sic) dysentery will take place at random in test is divided into two groups: one group of oral Bdellovibrio content is 10
6Pfu/ml, phage content are 10
10Pfu/ml mixed culture 10ml drug treatment, once a day, for three days on end; Another group is an oral ofloxacin 200mg drug treatment, once a day, and for three days on end.Then every day the observed and recorded piglet course of disease situation of change, continuous 10 days.
5, the results are shown in subordinate list 12,13.
Table 12. Bdellovibrio, phage mixed culture are to the preventive effect of yellow and white dysentery of piglet
Table 13. Bdellovibrio, phage mixed culture are to the therapeutic effect of yellow and white dysentery of piglet
6, conclusion (of pressure testing):
1), Bdellovibrio, phage mixed culture have the prevention protective effect to yellow and white dysentery of piglet, can improve the prevention protective rate with the ofloxacin group contrast is 82.63%;
2), Bdellovibrio, phage mixed culture have therapeutic effect to yellow and white dysentery of piglet, cure rate is 95.83%, can improve cure rate 18.91% with the ofloxacin group contrast;
3), Bdellovibrio, phage mixed culture are better than ofloxacin to the prevention effect of yellow and white dysentery of piglet.
Embodiment 13: Bdellovibrio, phage mixed culture are to poultry preventing and treating bacterial diseases of young effect measuring
1, bacterium source: Bdellovibrio, phage are provided by Chinese medicine antibacterial preservation administrative center vibrio phage specialized laboratory.
2, the screening of Bdellovibrio bacteriovorus bacterial strain and phage strain: from the Bdellovibrio bacteriovorus bacterial strain that obtains and phage strain, filter out Bdellovibrio Bd81, Bd98, Bd329, Bd334 and swine escherichia coli ∮ T101, Salmonella enteritidis ∮/50041 phagies to poultry common pathogen sensitivity with conventional method.
3, the preparation of Bdellovibrio, phage mixed culture: the bacterial strain that filters out with step 2 is prepared by the method for embodiment one.
4, experimental design: experiment is organized in cities such as Jiangsu Province's East Platform, Xinghua, Jiangdu, Gaoyou, Jiangning by Jiangsu Province bureau of animal husbandry, the district carries out.
Prophylactic tria, it is 10 that the piglet birth adopted the oral cavity to gavage a 5ml Bdellovibrio content same day
6Pfu/ml, phage content are 10
10The pfu/ml mixed culture was strengthened once in the time of ten days; It is 10 that chickling hatches the oral 0.5ml Bdellovibrio content of employing on same day drinking-water
6Pfu/ml, phage content are 10
10The pfu/ml mixed culture was strengthened once in the time of ten days; The matched group oral antibiotic.Observed and recorded poultry bacterial disease generation every day incidence then.Continuous 30 days.
Therapeutic test, test 1 is 10 with the oral Bdellovibrio content of the poultry that HUANGBAI(sic) dysentery takes place
6Pfu/ml, phage content are 10
10Pfu/ml mixed culture 10-30ml drug treatment, once a day, for three days on end; Test 2 is 10 with the oral Bdellovibrio content of the fowl that bacterial disease takes place
6Pfu/ml, phage content are 10
10Pfu/ml mixed culture 1.0-3.0ml drug treatment, once a day, for three days on end; Establish the matched group of oral antibiotic drug treatment simultaneously.The course of disease situation of change of 1 poultry of observed and recorded every day then, continuous 10 days.
5, the results are shown in subordinate list 14,15.
Table 14. Bdellovibrio, phage mixed culture are to the preventive effect of poultry bacterial disease
Table 15. Bdellovibrio, phage mixed culture are to the therapeutic effect of poultry bacterial disease
6, conclusion (of pressure testing):
1), Bdellovibrio, phage mixed culture are to the generation of poultry bacterial disease and popular the prevention protective effect arranged, protective rate is respectively 82.41%, 78.22%; Be higher than antibiotic 12.33%, 6.12% respectively.
2), Bdellovibrio, phage mixed culture have therapeutic effect to the poultry bacterial disease, cure rate is respectively 100%, 99.33%, is higher than antibiotic 10.68%, 22.23% respectively.
3), Bdellovibrio, phage mixed culture are better than antibiotic to the prevention effect of poultry bacterial disease.
Embodiment 14: Bdellovibrio, phage mixed culture are to the preventive effect of DISEASE IN FLOCKS
1, bacterium source: Bdellovibrio, phage are provided by Chinese medicine antibacterial preservation administrative center vibrio phage specialized laboratory.
2, the screening of Bdellovibrio bacteriovorus bacterial strain and phage strain: from the Bdellovibrio bacteriovorus bacterial strain that obtains and phage strain, filter out Bdellovibrio Bd76, Bd334 and Salmonella enteritidis ∮/50041 phagies to Pullorum Disease bacillus sensitivity with conventional method.
3, the preparation of Bdellovibrio, phage mixed culture: the bacterial strain that filters out with step 2 is prepared by the method for embodiment one.
4, experimental design: experiment is carried out in cities such as Yingkou, Liaoning Province, Panjin Fushun by Yingkou City, Liaoning Province Center for Disease Control (CDC) tissue.
It is 10 that chickling hatches the oral 0.5ml Bdellovibrio content of employing on same day drinking-water
6Pfu/ml, phage content are 10
10The pfu/ml mixed culture, continuous 7 days; Matched group oral antibiotic and blank group.Then every day the observed and recorded white diarrhea disease the generation incidence.
5, the results are shown in Table 16.
Table 16. Bdellovibrio, phage mixed culture are to the preventive effect of DISEASE IN FLOCKS
6, conclusion (of pressure testing):
1), Bdellovibrio, phage mixed culture be to the prevention protective effect of DISEASE IN FLOCKS, protective rate is 87.00%, antibiotic 71.00%.Bdellovibrio, phage mixed culture protective rate are higher than antibiotic 16.00%.
2), Bdellovibrio, phage mixed culture are better than antibiotic to the prevention protection effect of DISEASE IN FLOCKS.
Embodiment 15: Bdellovibrio, phage mixed culture are to the prevention effect of chick Hakuri
1, bacterium source: Bdellovibrio, phage are provided by Chinese medicine antibacterial preservation administrative center vibrio phage specialized laboratory.
2, the screening of Bdellovibrio bacteriovorus bacterial strain and phage strain: from the Bdellovibrio bacteriovorus bacterial strain that obtains and phage strain, filter out Bdellovibrio Bd76, Bd334 and Salmonella enteritidis ∮/50041 phagies to the common Pullorum Disease bacillus sensitivity of fowl with conventional method.
3, the preparation of Bdellovibrio, phage mixed culture: the bacterial strain that filters out with step 2 is prepared by the method for embodiment one.
4, experimental design: experiment is carried out in Agricultural University Of Nanjing.Test divides two groups, 45 of 45 Ai Wei mattresses of every group 1 age in days chickling.Test group, the oral 0.5ml Bdellovibrio content of drinking water every day is 10
6Pfu/ml, phage content are 10
10The pfu/ml mixed culture, continuous 4 days; Matched group need not any medicine.Two groups of feeding environments, management condition are identical.Continuous then 30 days, observed and recorded bacterial disease generation every day incidence.
5, the results are shown in Table 17.
Table 17. Bdellovibrio, phage mixed culture are to the preventive effect of DISEASE IN FLOCKS
6, conclusion (of pressure testing):
Bdellovibrio, phage mixed culture have tangible prevention protective effect to DISEASE IN FLOCKS, and protective rate is 100.00%.
Embodiment 16: Bdellovibrio, phage mixed culture are to the prevention effect of chick Hakuri
1, bacterium source: Bdellovibrio, phage are provided by Chinese medicine antibacterial preservation administrative center vibrio phage specialized laboratory.
2, the screening of Bdellovibrio bacteriovorus bacterial strain and phage strain: from the Bdellovibrio bacteriovorus bacterial strain that obtains and phage strain, filter out Bdellovibrio Bd76, Bd334, Bd329 and Salmonella enteritidis bacteriophage ∮/50041, coliphage ∮/T1 to the common Pullorum Disease bacillus sensitivity of fowl with conventional method.
3, the preparation of Bdellovibrio, phage mixed culture: the bacterial strain that filters out with step 2 is prepared by the method for embodiment one.
4, experimental design: experiment is carried out in Jiangning District chicken farm, Nanjing.Test divides two groups, 14600 of every group 1 red POLO of age in days, Ai Wei mattress chickling.Test group, the oral 0.5ml Bdellovibrio content of drinking water every day is 10
6Pfu/ml, phage content are 10
10The pfu/ml mixed culture, for three days on end; Matched group is with other antibacterials or without the medicine blank.Each organizes feeding environment, management condition is identical.Continuous then 56 days, observed and recorded bacterial disease generation every day incidence.
3, the results are shown in Table 18.
Table 18. Bdellovibrio, phage mixed culture are to the preventive effect of DISEASE IN FLOCKS
6, conclusion (of pressure testing):
Bdellovibrio, phage mixed culture have tangible preventive effect to DISEASE IN FLOCKS, with matched group comparison protective rate be 79.34%.
Embodiment 17: Bdellovibrio, phage mixed culture are measured the yellow and white dysentery of piglet therapeutic effect
1, bacterium source: Bdellovibrio, phage are provided by Chinese medicine antibacterial preservation administrative center vibrio phage specialized laboratory.
2, the screening of Bdellovibrio bacteriovorus bacterial strain and phage strain: from the Bdellovibrio bacteriovorus bacterial strain that obtains and phage strain, filter out Bdellovibrio Bd81, Bd98 and swine escherichia coli phage ∮ T101 to the swine escherichia coli sensitivity with conventional method.
3, the preparation of Bdellovibrio, phage mixed culture: the bacterial strain that filters out with step 2 is prepared by the method for embodiment one.
4, experimental design: carry out on the precious Huashan of the Nanjing Jiangning District live pig farm of raising pigs.Test is with 233 of the piglets in York hybridization breeding 20 ages in days, and test is divided into four groups: one group of oral Bdellovibrio content is 10
6Pfu/ml, phage content are 10
10Pfu/ml mixed culture 5ml, once a day, for three days on end; In addition two groups of oral gentamicin, chloromycetin 100mg, once a day, for three days on end.Establish simultaneously need not any medicine the blank group.Then every day the observed and recorded piglet course of disease situation of change, continuous 30 days.
5, the results are shown in Table 19.
Table 19. Bdellovibrio, phage mixed culture are to the preventive effect of yellow and white dysentery of piglet
6, conclusion (of pressure testing):
Bdellovibrio, phage mixed culture have preventive effect to yellow and white dysentery of piglet, and protective rate reaches 82.11%, and protective rate is higher than gentamycin 59.77%, chloromycetin 71.05%.
Embodiment 18: Bdellovibrio, phage mixed culture are to the plant pathogen lytic effect
1, bacterium source: Bdellovibrio, phage are provided by Chinese medicine antibacterial preservation administrative center vibrio phage specialized laboratory.Experiment is provided by agricultural product quality verification test center, Jiangsu Province with the plant pathogen bacterial strain.
2, the screening of Bdellovibrio bacteriovorus bacterial strain and phage strain: from the Bdellovibrio bacteriovorus bacterial strain that obtains and phage strain, filter out Bdellovibrio Bd81, Bd296, Bd334 and phage ∮ PE5, ∮ 1537, ∮ 119X, ∮ T7, ∮ 1214 to 9 strain plant pathogen sensitivities with conventional method.
3, the preparation of Bdellovibrio, phage mixed culture: the bacterial strain that filters out with step 2 is prepared by the method for embodiment one.
4, the preparation of plant pathogen culture: plant pathogen erwinia amylovora EA137, soft rot of cabbage bacterium EC153, the big pathogenic bacteria Tr1 of unwrapping wire vegetable fertilizer, Kidney bean aeruginosa atcc 11355, Solanaceae pseudomonas PS138, corynebacterium michiganense CM9, Nicotiana tabacum L. pseudomonas NRRLB877, bacillus pyocyaneus 2019, a blue or green pathogenic bacteria S173 separate with the nutrient agar panel line respectively, cultivated 18 hours for 37 ℃, select the colonies typical transferred species in the nutrient broth 37 ℃ cultivated 4-6 hour, promptly obtain the plant pathogen culture.
Experimental design: with Bdellovibrio content is 10
6Pfu/ml, phage content are 10
10The pfu/ml mixed culture is 10 to 9 strain plant pathogen content
8The cracking of cfu/ml culture.
Use the double-layer tap water agar flat band method, every flat board is the host with 3,000,000,000 cfu plant pathogens, adds 0.5ml Bdellovibrio and phage mixed culture simultaneously, cultivates 48 hours observed and recorded results in 37 ℃.Establishing escherichia coli C600 simultaneously is matched group, checks that Bdellovibrio, phage mixed culture form the ability of plaque.
5, the results are shown in Table 20.
Table 20. Bdellovibrio, phage mixed culture are to the lytic effect of plant pathogen
6, conclusion (of pressure testing):
Bdellovibrio, phage mixed culture all have splitting action to 9 strain plant pathogens, and cleavage rate is 100%.Infer that in view of the above Bdellovibrio, phage mixed culture have bigger using value to the control of bacteriosis.
Embodiment 19: Bdellovibrio, phage mixed culture are to the observation of blackberry antisepsis
1, bacterium source: Bdellovibrio, phage are provided by Chinese medicine antibacterial preservation administrative center vibrio phage specialized laboratory.
2, the screening of Bdellovibrio bacteriovorus bacterial strain and phage strain: from the Bdellovibrio bacteriovorus bacterial strain that obtains and phage strain, filter out Bdellovibrio Bd81, Bd296, Bd334 and phage ∮ PE5, ∮ 1537, ∮ 119X, ∮ T7, ∮ 1214 to the plant pathogen sensitivity with conventional method.
3, the preparation of Bdellovibrio, phage mixed culture: the bacterial strain that filters out with step 2 is prepared by the method for embodiment one.
4, experimental design: test in blackberry planting base, Jurong, Jiangsu Province county.Test divides two groups: one group is that to pluck preceding 24 hours in blackberry be 10 with nebulization with Bdellovibrio content
6Pfu/ml, phage content are 10
10The pfu/ml mixed culture evenly is sprayed at the blackberry surface that sophisticated plan is plucked, based on fruit.Pluck after 24 hours, pluck the conventional preservation in back; Two groups is will be 10 with Bdellovibrio content with nebulization at once after blackberry is plucked
6Pfu/ml, phage content are 10
10The pfu/ml mixed culture evenly is sprayed at the blackberry surface of having plucked, conventional then the preservation.Two groups of blanks of all establishing without Bdellovibrio, phage mixed culture.
5, the results are shown in Table 21.
Table 21. Bdellovibrio, phage mixed culture are to the observation of blackberry antisepsis
6, conclusion (of pressure testing):
1), pluck in blackberry and with nebulization Bdellovibrio, phage mixed culture evenly to be sprayed at the blackberry surface that sophisticated plan is plucked in preceding 24 hours, can prolong blackberry and guarantee the quality fresh-keeping 3-4 days;
2), after blackberry is plucked, with nebulization Bdellovibrio, phage mixed culture evenly are sprayed at the blackberry surface of having plucked at once, can prolong blackberry and guarantee the quality fresh-keeping 1-2 days.
Embodiment 20: Bdellovibrio, phage mixed culture are to the preventive and therapeutic effect of Rhizoma Zingiberis Recens and green pepper bacterial wilt
1, bacterium source: Bdellovibrio, phage are provided by Chinese medicine antibacterial preservation administrative center vibrio phage specialized laboratory.
2, the screening of Bdellovibrio bacteriovorus bacterial strain and phage strain: from the Bdellovibrio bacteriovorus bacterial strain that obtains and phage strain, filter out Bdellovibrio Bd81, the Bd296 of plant pathogen sensitivity and phage ∮ PE5, ∮ 1537, ∮ 119X with conventional method.
3, the preparation of Bdellovibrio, phage mixed culture: the bacterial strain that filters out with step 2 is prepared by the method for embodiment one.
4, experimental design: Vegetable Company is tested at the green home of Shandong Province's Laiwu City.Test divides two groups: one group is to select each 500 in the green pepper of just having emerged and Rhizoma Zingiberis Recens, is 10 with Bdellovibrio content
8Pfu/ml, phage content are 10
10Pfu/ml mixed culture 10ml pouring seedling root repeats once continuous then 50 days observed and recorded growing states after 20 days; Two groups is to select to fall ill or each 50 in the green pepper of initiation potential and Rhizoma Zingiberis Recens arranged, and is every milliliter 10 with Bdellovibrio content
6Pfu, phage content is 10 for every milliliter
10Pfu mixed culture 20ml waters root, and every day 1 time for three days on end.Two groups of green pepper and Rhizoma Zingiberis Recens seedling that all in equivalent environment, select same quantity, or the green pepper of morbidity and Rhizoma Zingiberis Recens are without the blank of Bdellovibrio, phage mixed culture.
5, the results are shown in Table 22,23.
Table 22. Bdellovibrio, phage mixed culture are to green pepper, the blue or green withered preventive effect of Rhizoma Zingiberis Recens
Table 23. Bdellovibrio, phage mixed culture are to the effect of green pepper, the blue or green withered treatment of Rhizoma Zingiberis Recens
6, conclusion (of pressure testing):
1), with Bdellovibrio, phage mixed culture pouring seedling root, the green pepper bacterial wilt is had preventive effect, protective rate is 82.35%;
2), with Bdellovibrio, phage mixed culture pouring seedling root, green pepper Rhizoma Zingiberis Recens disease is had preventive effect, protective rate is 73.68%;
3), with Bdellovibrio, phage mixed culture pouring seedling root, treatment is 85.71% (24/28) to the cure rate of green pepper bacterial wilt;
4), with Bdellovibrio, phage mixed culture pouring seedling root, treatment is 77.27% (17/22) to the cure rate of Rhizoma Zingiberis Recens bacterial wilt.
Embodiment 21: Bdellovibrio, phage mixed culture are observed coliform scavenging action in the river
1, bacterium source: Chinese medicine antibacterial preservation administrative center vibrio phage specialized laboratory provides.
2, the screening of Bdellovibrio bacteriovorus bacterial strain and phage strain: from the Bdellovibrio bacteriovorus bacterial strain that obtains and phage strain, filter out Bdellovibrio Bd98, Bd59, Bd329 and coliphage ∮/T1, ∮/T2, ∮/T7 to the coliform sensitivity with conventional method.
3, the preparation of Bdellovibrio, phage mixed culture: the bacterial strain that filters out with step 2 is prepared by the method for embodiment one.
4, experimental design: in natural river, the content that adds Bdellovibrio is 10
4Pfu/ml, the content of phage are 10
6The pfu/ml mixed culture.Establish simultaneously and do not add Bdellovibrio, phage mixed culture matched group.The timing water sampling detects the variation of coliform in the water body later on.Continue 33 days.
5, the results are shown in Table 24.
Table 24. Bdellovibrio, phage mixed culture are to the influence of coliform in the river
6, conclusion (of pressure testing):
Test group and matched group compare, and statistical procedures (P＜0.01) has significant difference, thereby prove that Bdellovibrio, phage mixed culture have scavenging action to coliform in the river.
Embodiment 22: Bdellovibrio, phage mixed culture are observed total number of bacteria scavenging action in the river
1, bacterium source: Chinese medicine antibacterial preservation administrative center vibrio phage specialized laboratory provides.
2, the screening of Bdellovibrio bacteriovorus bacterial strain and phage strain: from the Bdellovibrio bacteriovorus bacterial strain that obtains and phage strain, filter out Bdellovibrio Bd59, Bd81, Bd98, Bd329 and vibrio phage ∮ V3, ∮ M6 to pathogenic bacterium sensitivity common in the river with conventional method, shigella phage ∮ P22, typhoid fever phage ∮ Vi, ∮ S154, Escherichia coli phage ∮ T2, ∮ T7, pyocinophages ∮ PE5.
3, the preparation of Bdellovibrio, phage mixed culture: the bacterial strain that filters out with step 2 is prepared by the method for embodiment one.
4, experimental design: in natural river, the content that adds Bdellovibrio is 10
4Pfu/ml, the content of phage are 10
5The pfu/ml mixed culture.Establish simultaneously and do not add Bdellovibrio, phage mixed culture matched group.The timing water sampling detects the variation of total number of bacteria in the water body later on.Continue 30 days.
5, the results are shown in Table 25.
Table 25. Bdellovibrio, phage mixed culture are to total number of bacteria in the river
6, conclusion (of pressure testing):
Test group and matched group compare, and the amplitude that total number of bacteria reduces in the river of Bdellovibrio, phage mixed culture is higher than matched group 39-55 doubly.
Embodiment 23: Bdellovibrio, phage mixed culture are to the observation of cyanophyceae control action in the river
1, bacterium source: Chinese medicine antibacterial preservation administrative center vibrio phage specialized laboratory provides.
2, the screening of Bdellovibrio bacteriovorus bacterial strain and phage strain: from the Bdellovibrio bacteriovorus bacterial strain that obtains and phage strain, filter out Bdellovibrio Bd59, Bd296 and phage ∮ 53K, ∮ S157 to the plant pathogen sensitivity with conventional method.
3, the preparation of Bdellovibrio, phage mixed culture: the bacterial strain that filters out with step 2 is prepared by the method for embodiment one.
4, experimental design: send out in Sheyang County, Jiangsu Province farm Aquatic product (channel catfishes) cultivation base positive and test.The Ietalurus Punetaus of test and Selection cyanophyceae heavy contamination is cultured 10 in fish pond, and the area on each pool is 150m * 300m, the about 1.2m of the depth of water.Test group was splashed in per 10 days, and (content is 10 to 1 Bdellovibrio
8/ pfu), (content is 10 to phage
10/ pfu) mixed culture 300ml/ mu/rice.Continuous 3 times.Continuous then 90 days observed and recorded blue algae growth situations; Select the heavier Ietalurus Punetaus of equivalent environment medium blue algae pollution to culture 5 in fish pond, without Bdellovibrio, phage mixed culture and with the matched group of quicklime.
5, the results are shown in Table 26.
Table 26. Bdellovibrio, phage mixed culture are to cyanophyceae control observation on effect in the river
6, conclusion (of pressure testing):
Bdellovibrio, phage mixed culture have very obvious control effect to spreading of cyanophyceae in the river.
Claims (2)
1. an active bio antibacterial is characterized in that, is that culture with Bdellovibrio and phage is that Main Ingredients and Appearance is mixed in proportion and makes, and it is characterized in that the mixed culture of Bdellovibrio and phage prepares by the following method:
(1) acquisition of Bdellovibrio culture: the Bdellovibrio host bacterium that makes inactivation earlier, be the host with this inactivation host bacterium then, produce the Bdellovibrio culture with the Bdellovibrio general culture method, select on the double-deck agar plate of tap water plaque typical again, energy cracking noxious bacteria, do not destroy beneficial bacteria, the Bdellovibrio bacteriovorus bacterial strain that in 10-43 ℃ of environment, can grow, at last inactivation host bacterium that makes in the said process and the Bdellovibrio that the selects ratio in 10:1-10000:1 is suspended in the fermentation culture, proofread and correct its pH value 3.8-10.0, through fermentation culture, fermentation culture medium or with standby after the fermentation culture medium lyophilizing;
(2) acquisition of phage culture: produce responsive phage host bacteria suspension, produce the phage culture with the double-deck agar culture method of phage, select the virulent phage of the typical energy of plaque cracking noxious bacteria on the double-deck agar plate of nutrient broth again, single plaque that the picking transparency is good is soaked in the nutrient broth, then, get supernatant and make the double-deck agar plate of nutrient broth again, at last with the phage culture that makes in the said process and responsive host bacteria suspension by 1:10-1:10
8Ratio be suspended in the fermentation culture, proofread and correct its pH value 3.8-10.0, through fermentation culture, fermentation culture medium is removed responsive host bacterium through aseptic filtration; Determine aseptic fermentation culture medium or the fermentation culture medium lyophilizing is standby through checking;
(3) with above-mentioned Bdellovibrio fermentation culture medium or with fermentation culture medium filtering and concentrating lyophilized powder with through checking the phage fermentation culture medium after determining aseptic aseptic filtration or with the ratio preparation of fermentation culture medium lyophilized powder in 1:1-1:1000000, promptly the content of Bdellovibrio is every milliliter of 10-10
10Pfu, the content of phage is 10-10 for every milliliter
16Pfu; Mixed preparing makes it be suspended in medium liquid or is blended in the excipient; Promptly obtain product.
2. a method for preparing the described active bio antibacterial of claim 1 is characterized in that, may further comprise the steps:
(1) acquisition of Bdellovibrio culture: the Bdellovibrio host bacterium that makes inactivation earlier, be the host with this inactivation host bacterium then, produce the Bdellovibrio culture with the Bdellovibrio general culture method, select on the double-deck agar plate of tap water plaque typical again, energy cracking noxious bacteria, do not destroy beneficial bacteria, the Bdellovibrio bacteriovorus bacterial strain that in 10-43 ℃ of environment, can grow, at last in the fermentation culture that the inactivation host bacterium that makes in the said process and the Bdellovibrio that selects are suspended in the ratio of 10:1-10000:1, proofread and correct its pH value, through fermentation culture, fermentation culture medium or with standby after the fermentation culture medium lyophilizing;
(2) acquisition of phage culture
Produce responsive phage host bacteria suspension, produce the phage culture with the double-deck agar culture method of phage, select the virulent phage of the typical energy of plaque cracking noxious bacteria on the double-deck agar plate of nutrient broth again, single plaque that the picking transparency is good is soaked in the nutrient broth, then, get supernatant and make the double-deck agar plate of nutrient broth again, at last with the phage culture that makes in the said process and responsive host bacteria suspension by 1:10-1:10
8Ratio be suspended in the fermentation culture, proofread and correct its pH value, through fermentation culture, fermentation culture medium is removed responsive host bacterium through aseptic filtration; Determine aseptic fermentation culture medium or the fermentation culture medium lyophilizing is standby through checking;
(3) determine aseptic phage fermentation culture medium or with the ratio preparation of fermentation culture medium lyophilized powder in 1:1-1:1000000, promptly the content of Bdellovibrio is every milliliter of 10-10 with above-mentioned Bdellovibrio fermentation culture medium or with the fermentation culture medium lyophilized powder with through checking
10Pfu, the content of phage is 10-10 for every milliliter
16Pfu; Mixed preparing makes it be suspended in medium liquid or is blended in the excipient; Promptly obtain product.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1090761A (en) * | 1993-09-22 | 1994-08-17 | 秦生巨 | Bactericide prepared from biology and production method thereof |
| CN1857327A (en) * | 2006-04-06 | 2006-11-08 | 薛恒平 | Production process of bdellophage preparation |
-
2010
- 2010-11-18 CN CN201010549139A patent/CN101991611B/en not_active Expired - Fee Related
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1090761A (en) * | 1993-09-22 | 1994-08-17 | 秦生巨 | Bactericide prepared from biology and production method thereof |
| CN1857327A (en) * | 2006-04-06 | 2006-11-08 | 薛恒平 | Production process of bdellophage preparation |
Non-Patent Citations (2)
| Title |
|---|
| 《生物学杂志》 19871018 罗成等 噬菌体研究的历史与现状 第20-23页 1-2 , 第19期 2 * |
| 《贵阳医学院学报》 20091231 曾佳 噬菌蛭弧菌的分离培养研究 第689-690页 1-2 第34卷, 第6期 2 * |
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