Method for counting effective viable bacterium content of Bdellovibrio bacteriovorus powder

A technology for the content of Bdellovibrio and live bacteria, which is applied in the field of microbial determination, can solve problems such as reduction, inability to distinguish dead bacteria from live bacteria, and inaccurate detection results of Bdellovibrio accurate effect

Active Publication Date: 2012-02-15
绿奥环保科技(上海)有限公司
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Problems solved by technology

For example, the microscope direct counting method cannot distinguish dead bacteria from live bacteria, and the counting result is the sum of dead bacteria and live bacteria; the dilute tap water double-layer agar plate me

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  • Method for counting effective viable bacterium content of Bdellovibrio bacteriovorus powder

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Embodiment 1

[0011] 1 Materials and methods

[0012] 1.1 Experimental materials

[0013] Phage Bdellovibrio powder BBM, the effective live bacteria content determined by tap water double-layer agar plate method is 3×10 7 pfu / g, purchased from Shanxi Weishi Yukang Industrial Co., Ltd.; 1mL glass pipette, sterile glass coating rod, 1.5% tap water agar plate, 0.6% tap water soft agar (incubated at 65°C), nutrient broth , 50mL stoppered centrifuge tube, slide glass, purchased from Sinopharm Chemical Reagent Co., Ltd.; Escherichia coli, purchased from TaKaRa Company; 1mL glass pipette, nutrient broth, filled with 9mL 10-fold diluted sterile nutrient broth 50mL stoppered centrifuge tube, 0.6% tap water soft agar (incubated at 65°C), sterilized at 121°C for 20min; biochemical incubator, purchased from Japan Sanyo Company; vortex oscillator, purchased from Shanghai Qite Analytical Instrument Co., Ltd. Company; temperature-controlled shaker, purchased from the United States Forma Scientific compa...

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Abstract

The invention provides a method for counting the effective viable bacterium content of Bdellovibrio bacteriovorus powder, which comprises the following steps: appropriately diluting a sample to be detected, so that Bdellovibrio bacteriovorus in the sample to be detected is sufficiently dispersed into individual cells; then, carrying out incubation culture in an aseptic nutrient broth diluted by 10 times to promote growth and propagation, thus forming a megascopic turbid liquid; further determining that a large amount of Bdellovibrio bacteriovorus is contained in the turbid liquid having the maximum dilution through plaque detection; and finally, determining the total amount of effective viable bacteria in the sample to be detected. Compared with a microscopic direct observation method anda dilution-by-tap-water double-layer agar plate method, the method provided by the invention is more accurate, and overcomes the defects that detection is inaccurate due to dilution in the dilution-by-tap-water double-layer agar plate method and killed bacteria and viable bacteria can not be distinguished in the microscopic direct observation method.

Description

technical field [0001] The invention relates to a microorganism measurement technology, in particular to a method for counting the effective viable bacteria content of bacteriophage Bdellovibrio powder. Background technique [0002] Bdellovibrio phage preparation is a green product with no toxic side effects and no drug residues. It is rich in Bdellovibrio phage with unique phage characteristics, and is a commonly used water quality microbial improver in aquaculture. However, the lack of quality standards is one of the main problems in the development of bacteriophage Bdellovibrio preparations, and one of the biggest problems leading to the lack of quality standards is that the number of live bacteria does not match the label, and some live bacteria are even difficult to detect. Therefore, a stable and reliable counting method of Bdellovibrio preparations is undoubtedly of great significance to the normal production and quality control of Bdellovibrio preparations. At prese...

Claims

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Application Information

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IPC IPC(8): C12Q1/08C12R1/63
Inventor 曹海鹏何珊卫若鹏
Owner 绿奥环保科技(上海)有限公司
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