CN1076755C - Qinghai Vibrion dry-powder preparation - Google Patents
Qinghai Vibrion dry-powder preparation Download PDFInfo
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- CN1076755C CN1076755C CN97106203A CN97106203A CN1076755C CN 1076755 C CN1076755 C CN 1076755C CN 97106203 A CN97106203 A CN 97106203A CN 97106203 A CN97106203 A CN 97106203A CN 1076755 C CN1076755 C CN 1076755C
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- qinghai
- qinghai vibrion
- resuscitation fluid
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Abstract
The present invention discloses a dry powder preparation of Qinghai vibrio, which is formed in a mode that Qinghai vibrio is cultured, screened and added with protection liquid to be made into a dry powder preparation. Different resuscitation fluids are adopted to reanimate the Qinghai vibrio in use according to tested objects with different water qualities to test the biologic toxicity of the water qualities.
Description
Qinghai Vibrion of the present invention (Vibrio qinghaiensis) dry powder formulations microorganism belonging to genus formulation art.
At present the test of bio-toxicity to be had several different methods but be simple be to adopt ocean photogenic bacterium (mainly being photobacterium phosphoreum) to carry out to water quality, as Bulich.A.A at AmericanSociety for Testing and materials, Philadolphia, PA, 1979, described in the PP98-106 article like that,, the luminous physiological requirement of ocean luminescent bacteria is relatively harsher, it must contain 3%NaCl, and acidity is about PH7, and the temperature of environment is controlled between 15-25 ℃, in environment so, can show its normal luminous instinct.But in practical measurement,, the confidence level that records the result is reduced, the utilization of ocean luminescent bacteria in test organisms toxicity is restricted widely owing to after in pollution water sample, adding 3%NaCl, the bio-toxicity of sample is changed.
The present invention's ocean luminescent bacteria restriction of being subjected on the bio-toxicity of test water quality for a change; find that in practice the luminous physiological requirement of Qinghai Vibrion is lower than the luminous physiological requirement of ocean luminescent bacteria; advantage applied widely (sees red legend outstanding person etc. for details, in May, 1994, photogenic bacterium one novel species-Qinghai Vibrion; Oceanologia et Limnologia Sinica; the 25th volume, the 3rd phase, PP273-279); and Qinghai Vibrion can be made into dry powder doses, makes it easier preservation and application.
The present invention utilize naked carp in Qinghai Lake the body surface culture of isolated and photogenic bacterium--Qinghai Vibrion; this Qinghai Vibrion is made dry powder formulations; its major technique be with Qinghai Vibrion through liquid culture to logarithmic phase; the vibrios body is collected in centrifugation, then with after the thalline washes clean, is suspended in again in the protective material solution; the prescription of its protective material solution is milk 1-10%wt; sucrose or trehalose 1-10%wt, arginine 0.1-0.5%wt, water are surplus.Suspension with thalline is sub-packed in ampoule again, and every bottle of 1ml places ice to make it to be frozen into solid-state, then, takes out and to place that to be dried to water content on the freeze drier about about 5%, and with ampule sealing, it is stand-by below 10 ℃ to store in refrigerator.In test; at first make Qinghai Vibrion dry-powder preparation add resuscitation fluid A or resuscitation fluid B; after making its recovery; toxicity to water quality is tested; its resuscitation fluid A is the phosphoric acid buffer that volumetric molar concentration and acidity are respectively 0.05-0.2M and PH6.0-9.0, and resuscitation fluid B is that percentage concentration is the 0.5-8% Glucose Liquid.
Qinghai Vibrion stabilized illumination more than hour in the distilled water environment, still continued growth and luminous good in the 5%NaCl saline solution.Growth that adapts to and luminous temperature and acidity are respectively in the scope of 10-30 ℃ and PH5.5-9.5.These growths of Qinghai Vibrion, the characteristics of luminescence is better than the ocean photogenic bacterium greatly to growth, luminous environmental requirement.Therefore, the acidity that Qinghai Vibrion uses, temperature, salinity wider general, the result of mensuration is more reliable.
Embodiment:
With the Qinghai Vibrion liquid culture to mid-log phase; with the whizzer of 3000rpm centrifugal 10 minutes; collect thalline; thalline is suspended in protective material solution with distilled water wash after clean again; its protective material solution is milk; trehalose; arginine and water, its concentration is respectively 8%, 4% and 0.3%; surplus is a water; then Qinghai Vibrion suspension is sub-packed in ampoule, every bottle is 1ml, and it is solid-state to place dry ice that it is frozen into; putting into after the taking-up again and being dried to water ratio on the freeze drier is about 5%, places refrigerator stand-by below 10 ℃.
In testing sample, add NaCl; making its salinity is 0.5%NaCl, or adds glucose, and its concentration is about 3%; take out Qinghai Vibrion dry-powder preparation then; add 3ml resuscitation fluid A in ampoule, shake up, leave standstill 5min; resuscitation fluid A is the 0.07MPH9.0 phosphoric acid buffer; Qinghai Vibrion 50l after the recovery is added in the 2ml analyte sample fluid, place on the luminous photometer and measure luminous intensity, make blank solution luminous intensity test controlled trial simultaneously.Try to achieve the degree of water quality toxicity according to this with luminous inhibiting rate.
2. get Qinghai Vibrion dry-powder preparation, handle through resuscitation fluid B, resuscitation fluid B is 2% glucose, the aqueous solution.In the testing sample of 2ml, (containing 4% glucose) adds 50l resuscitation fluid B Qinghai Vibrion must, and the method for pressing embodiment 1 is carried out same mensuration, tries to achieve the water quality toxicity based on heavy metal contamination according to this.
Claims (3)
1. a Qinghai Vibrion (Vibrio qinghaiensis) dry powder formulations; Qinghai Vibrion be a kind of body surface that picks up from naked carp in the Qinghai Lake, through culture of isolated and fresh water type photogenic bacterium; it is characterized in that, the said preparation obtained by freeze drying that is Qinghai Vibrion under the protective material solution protection that contains milk, sucrose or trehalose, arginine and water, water content is 5% dry powder formulations.
2. the method for making of the described Qinghai Vibrion of claim 1 (Vibrio qinghaiensis) dry powder formulations; with the Qinghai Vibrion that picks up from the body surface of naked carp in the Qinghai Lake through liquid culture to mid-log phase; thalline is collected in centrifugation; and with the thalline washes clean; it is characterized in that; clean thalline is suspended in protective material solution; thallus suspension liquid is sub-packed in ampoule; every bottle of 1ml; place dry ice to make it to be frozen into solid-state this ampoule, then, take out and to place that to be dried to water content on the freeze drier be 5%; with ampule sealing; store in the refrigerator below 10 ℃, stand-by, the solid matter in this ampoule is finished product; Qinghai Vibrion dry-powder preparation, the prescription of above-mentioned protective material solution is:
Milk 1-10%wt
Sucrose or trehalose 1-10%wt
Arginine 0.1-0.5%wt
Water surplus
3. the purposes of the described Qinghai Vibrion of claim 1 (Vibrio qinghaiensis) dry powder formulations, utilize the bio-toxicity of described preparation test water quality, in testing sample, add NaCl or grape glucose, be respectively 0.5% or 2-4% to the per-cent salinity of this sample or percentage concentration, add the 3ml resuscitation fluid to the ampoule that fills described preparation, shake up, leave standstill 5min, the preparation 50 μ l of recovery are added in the testing sample of 2ml, place the luminous intensity of measuring thalline on the luminous photometer, make blank solution luminous intensity test controlled trial simultaneously, try to achieve the bio-toxicity of testing sample according to luminous inhibiting rate, it is characterized in that above-mentioned resuscitation fluid has two kinds of resuscitation fluid A and resuscitation fluid B: resuscitation fluid A is the phosphoric acid buffer that volumetric molar concentration and acidity are respectively 0.05-0.2M and pH6.0-9.0; Resuscitation fluid B is that percentage concentration is the Glucose Liquid of 0.5-8.0%.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN97106203A CN1076755C (en) | 1997-01-07 | 1997-01-07 | Qinghai Vibrion dry-powder preparation |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN97106203A CN1076755C (en) | 1997-01-07 | 1997-01-07 | Qinghai Vibrion dry-powder preparation |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1187537A CN1187537A (en) | 1998-07-15 |
| CN1076755C true CN1076755C (en) | 2001-12-26 |
Family
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Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN97106203A Expired - Lifetime CN1076755C (en) | 1997-01-07 | 1997-01-07 | Qinghai Vibrion dry-powder preparation |
Country Status (1)
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| CN (1) | CN1076755C (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102604873A (en) * | 2012-03-28 | 2012-07-25 | 北京滨松光子技术股份有限公司 | Rejuvenation method of marine luminous bacteria |
Families Citing this family (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103149199B (en) * | 2012-12-25 | 2015-05-27 | 北京城市排水集团有限责任公司 | Manufacturing method of water-quality acute toxicity high throughput rapid determination reagent kit and using method thereof |
| CN105368735B (en) * | 2013-01-18 | 2019-01-22 | 力合科技（湖南）股份有限公司 | Vibrio-qinghaiensis sp. Q67 B and its separation screening and application |
| CN104419644B (en) * | 2013-09-10 | 2018-07-06 | 上海市刑事科学技术研究院 | A kind of photobacteria lyophilized preparation and preparation method thereof |
| CN103468572B (en) * | 2013-09-25 | 2016-07-06 | 北京尚洋东方环境科技有限公司 | A kind of freeze drying protectant of luminescent bacteria |
| CN104762209B (en) * | 2015-03-30 | 2017-12-29 | 华东理工大学 | Photobacterium phosphoreum freeze-dried powder and preparation method thereof |
| CN106546579A (en) * | 2016-10-28 | 2017-03-29 | 华南理工大学 | A kind of organic solvent improves the method that photobacteria detects toxicant susceptibility |
| CN111004759B (en) * | 2020-01-04 | 2023-04-07 | 广东环凯生物科技有限公司 | Stabilizer for vibrio cholerae and application thereof |
-
1997
- 1997-01-07 CN CN97106203A patent/CN1076755C/en not_active Expired - Lifetime
Non-Patent Citations (3)
| Title |
|---|
| AM348CAN SOCIETY FOR TESTIJG AND MATERIALS HILADOLPHIA PA 1979.1.1 BULICH,A A * |
| AM348CAN SOCIETY FOR TESTIJG AND MATERIALS HILADOLPHIA PA 1979.1.1 BULICH,A A;AMAMICAMSONPA 1979.1.1 BUICHAA * |
| AMAMICAMSONPA 1979.1.1 BUICHAA * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102604873A (en) * | 2012-03-28 | 2012-07-25 | 北京滨松光子技术股份有限公司 | Rejuvenation method of marine luminous bacteria |
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| Publication number | Publication date |
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| CN1187537A (en) | 1998-07-15 |
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Owner name: BEIJING BINSONG PHOTON TECHNOLOGY CO., LTD. Free format text: FORMER OWNER: HUADONG NORMAL UNIV; SHANGHAI CITY ENVIRONMENT PROTECTION BUREAU Effective date: 20081010 |
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Effective date of registration: 20081010 Address after: Room 708, block A, silver net center, No. 113, Haidian District, Beijing, Zhichun Road: 100086 Patentee after: Beijing Hamamatsu Photonics Technology Co., Ltd. Address before: No. 3663, Shanghai, Zhongshan North Road: 200062 Co-patentee before: Shanghai Environmental Protection Bureau Patentee before: East China Normal University |
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Address after: 100070, No. 18, No. 11, No. 188, South Fourth Ring Road, Fengtai District, Beijing Patentee after: Beijing Hamamatsu Photonics Technology Co., Ltd. Address before: 100086, room 708, block A, silver net center, No. 113, Haidian District, Beijing, Zhichun Road Patentee before: Beijing Hamamatsu Photonics Technology Co., Ltd. |
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Address after: 100070, room 3, building 128, No. 903 South Fourth Ring Road, Beijing, Fengtai District Patentee after: Beijing Hamamatsu Photonics Technology Co., Ltd. Address before: 100070, No. 18, No. 11, No. 188, South Fourth Ring Road, Fengtai District, Beijing Patentee before: Beijing Hamamatsu Photonics Technology Co., Ltd. |
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Effective date of registration: 20160630 Address after: 065001, No. 21, No.1 Road, Langfang Development Zone, Hebei province (Room 101 of complex building) Patentee after: HAMAMATSU PHOTONICS MEDICAL TECHNOLOGY (LANGFANG) CO., LTD. Address before: 100070, room 3, building 128, No. 903 South Fourth Ring Road, Beijing, Fengtai District Patentee before: Beijing Hamamatsu Photonics Technology Co., Ltd. |
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