Method for separating pediococcus pentosaceus

A technology of Pediococcus pentosaceae and sterilizers, applied in the direction of microorganism-based methods, separation of microorganisms, biochemical equipment and methods, etc., can solve the problems of increasing the workload of identification work, reducing the probability of isolation of Pediococcus pentosaceae, and achieving The effect of reducing workload

Inactive Publication Date: 2012-01-25
INST OF SUBTROPICAL AGRI CHINESE ACAD OF SCI
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Problems solved by technology

Because Pediococcus pentosaceae belongs to facultative anaerobic bacteria, the growth of Pediococcus pentosaceae can be inhibited under the situation of too much oxygen, therefore, this method may reduce the isolati

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  • Method for separating pediococcus pentosaceus

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Embodiment 1

[0028] The following examples are used to illustrate the present invention, but are not intended to limit the scope of the present invention.

[0029] Below in conjunction with accompanying drawing, the present invention is described in further detail:

[0030] A method for isolating Pediococcus pentosaceae, the steps are:

[0031] A. MRS solid medium preparation: The medium formula is (parts by weight): beef protein powder 10, fish juice 10, yeast extract juice powder 5, glucose 20, sodium acetate 5, diammonium citrate 2, magnesium sulfate 0.58, manganese sulfate 0.28, agar powder 15, add 1 ml Tween 80, dissolve in distilled water 1000, adjust the pH value to 6 or 6.2 or 6.3 or 6.4 or 6.6, put it in a sterilizing pot at 115 ℃ for 13 or 14 or 15 or 16 or 17 minutes, naturally cooled to 40 or 42 or 43 or 44 or 45 ℃.

[0032] B. Sample preparation: Take 1 ml of porcine intestinal digesta, mix it with sterile double-distilled water at a volume ratio of 1:10, vortex for 1 minute...

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Abstract

The invention discloses a method for separating pediococcus pentosaceus, which comprises the following steps: A, regulating the pH of a prepared MRS (de Man, Rogosa and Sharpe) solid culture medium, and then, placing the MRS solid culture medium in a sterilizing pot for sterilizing, and naturally cooling the MRS solid culture medium; B, mixing a sample to be separated by using sterile double distilled water, whirling the mixture, and then standing the mixture; C, removing to obtain a supernatant fluid, thinning the supernatant fluid, respectively taking three dilution degrees of 10 to the power of -3, 10 to the power of -4 and 10 to the power of -5, respectively mixing the mixture with the prepared liquid MRS solid culture medium, adding the mixture in a glass plate, and shaking a culturing dish, so that the culture medium and a sample mixture are distributed at the bottom of the culture dish; and D, after the culture medium and the sample mixture are solidified, adding the MRS solid culture medium, solidifying the mixture, and then placing the mixture in a biochemical incubator to culture, so that the pediococcus pentosaceus is separated at the bottom of the culture dish. The method is feasible, and is simple and convenient to operate; after the sample to be separated is thinned, a diluent is poured into the plate; after the diluent is solidified, the diluent is poured into the culture medium to form an anaerobic environment; the probability that separated bacteria are cloned into the pediococcus pentosaceus is 100%; the method is more accurate; and the workload is reduced obviously.

Description

technical field [0001] The invention relates to a method for isolating Pediococcus pentosacea, in particular to a method suitable for isolating Pediococcus pentosacea from the gastrointestinal tract of animals. Background technique [0002] The Latin scientific name of Pediococcus is Pediococcus Claussen. The cells are spherical and never elongated, with a diameter of 1.2-2.0 microns; under suitable conditions, they divide in two directions to form quadruples. Single cells are rare and do not form chains; Lamb's stain is positive; no movement, no spore production; facultative anaerobic, some strains will inhibit growth when there is oxygen; the optimum growth temperature is 25-40 ℃. [0003] Pediococcus pentosacea ( Pediococcus pentosaceus ) is a kind of Pediococcus, the colony is round, milky white, smooth, raised, neatly edged, opaque, and Gram-stain positive. The Chinese Ministry of Agriculture has approved that Pediococcus pentosacea can be used as a microbial strain...

Claims

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Application Information

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IPC IPC(8): C12R1/01C12N1/02C12N1/20
Inventor 王升平印遇龙李丽立姚康黄瑞林李铁军李凤娜
Owner INST OF SUBTROPICAL AGRI CHINESE ACAD OF SCI
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