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Method for quickly determining bacterial number in Riemerella anatipestifer culture

A technique for cultivating Riemer's and bacilli in duck plague, applied in biochemical equipment and methods, measurement of color/spectral characteristics, measurement/inspection of microorganisms, etc., can solve problems such as the indetermination of the number of bacteria, shorten detection time, and operate Sophisticated, easy-to-use effects

Inactive Publication Date: 2011-01-05
SICHUAN AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the optical density or transmittance is not only affected by the concentration of bacteria, but also affected by factors such as cell size, shape, culture medium composition, and the wavelength of light used, and usually only detects suspensions containing a large number of bacteria. The relative number of bacteria can not be determined by this method for samples or tissue samples that are too dark in color, but this method is simple and quick
The main key to the determination of the number of bacteria by turbidimetry is how to eliminate the interference in the sample, the determination of the wavelength of the light wave used and the corresponding relationship between the measured OD value and the number of bacteria, otherwise only the relative number of bacteria can be judged but not determined accurate count of bacteria

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] ① Sampling: Direct quantitative sampling of liquid culture or bacterial liquid; ② Centrifugal washing: After centrifuging at 3000-5000 rpm for 5-10 minutes, wash with normal saline continuous centrifugation (3000-5000 rpm, 5-10 minutes) 3 times; ③Dilution: Dilute appropriately with normal saline; ④Detection: Measure the OD value at 560nm under the ultraviolet spectrophotometer (note that use normal saline to adjust to 0); The total number of bacteria (CFU / ml) = OD 560nm Value × dilution factor × 2 × 10 9 CFU / ml.

[0021] For example: after serum type I Riemerella anatipestifer RA-CH-I strain was fermented with N-J synthetic medium at 35-40°C for 24 hours, the number of bacteria contained in Riemerella anatipestifer in the fermentation liquid was detected, and the specific steps were as follows: :

[0022] ①Sampling: Take 1ml of fermentation broth; ②Centrifugal washing: After centrifuging at 3000-5000 rpm for 5-10 minutes, wash with normal saline continuously (3000-50...

Embodiment 2

[0025] This example lists a concrete operation example of solid culture.

[0026] For example: Serum Type I Riemerella anatipestifer RA-CH-I strain is cultivated in a 12em petri dish in a candle jar at 35-40°C for 24-48 hours with trypticase soybean agar medium, then rinse the duck plague in the petri dish For other researches on Bacillus mercii, if it is necessary to detect the number of bacteria, the specific steps are as follows:

[0027] ① Sampling: Add 5ml of normal saline to the petri dish, use a pipette to absorb the normal saline to rinse the Riemerella anatipestifer growing on the surface of the agar, pour the washed bacteria solution into a 15ml centrifuge tube; add another 5ml of normal saline Wash the agar plate repeatedly once to obtain a total of 10ml of bacterial liquid; ②Centrifugal washing: after centrifuging at 3000-5000 rpm for 5-10 minutes, wash with normal saline continuously (3000-5000 rpm, 5-10 minutes) 3 times; ③ Dilution: Dissolve the precipitate with...

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PUM

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Abstract

The invention discloses a method for quickly determining a bacterial number in a Riemerella anatipestifer culture, comprising the following steps of: (1) sampling; (2) centrifugal cleaning: after centrifuging for 5-10 minutes by 3000-5000 turns per minute, continuously and centrifugally washing 3 times by using physiological saline, wherein each time of centrifugal cleaning spends 5-10 minutes at rotation speed of 3000-5000 turns per minutes; (3) diluting: diluting N times by using the physiological saline; (4) detecting: detecting an OD (Outside Diameter) value at a 560 nm point under an ultraviolet spectrophotometer; and (5) calculating: calculating the number of Riemerella anatipestifer according to the following formula: a total bacterial number (CFU / ml)=an OD560nm value*2*109CFU (Clonal Formation Unit) / ml.

Description

technical field [0001] The invention relates to the counting of bacteria of Riemerella anatipestifer, in particular to a method for detecting the bacteria quantity of Riemerella anatipestifer in a Riemerella anatipestifer culture by using an ultraviolet spectrophotometer. Background technique [0002] Duck infectious serositis is a contagious disease that occurs in domestic ducks, geese, turkeys and a variety of poultry. Syndrome, duck plague septicemia and new duck disease, the pathogen is Riemerella anatipestifer (Riemerella anatipestifer, RA). The disease presents as acute sepsis or chronic infection. Clinically, the main manifestations are cough, panting, diarrhea, increased eye and nose secretions, ataxia, and head and neck tremor. Perihepatitis, air sacculitis, meningitis, and caseous salpingitis. The disease can occur all year round, and it is serious in winter, mainly through contaminated feed, drinking water, droplets, etc.; 1-8 week-old ducks are highly sensitive...

Claims

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Application Information

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IPC IPC(8): G01N21/33C12Q1/06
Inventor 程安春汪铭书朱德康陈孝跃
Owner SICHUAN AGRI UNIV
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