Summary of the invention:
In view of above present situation, the invention provides a kind of selective coloration culture medium of clostridium perfringens, the incubation time that has solved above-mentioned traditional culture identification method existence is long, lacks the problem of specificity.
Technical scheme of the present invention realizes by following content:
A kind of selective coloration culture medium of clostridium perfringens, it is characterized in that: the composition of configuration 1000ml substratum is casease hydrolyzate 12.0g-20.0g, yeast soaks powder 6.0g-12.0g, S-WAT 0.20g-0.80g, PXB 0.005g-0.015g, Sulphadiazine Sodium 0.05g-0.20g, ironic citrate 0.20g-0.80g, 4-12 unit of pancreas Suning, agar powder 10.0g-18.0g, adding distil water is to 1000ml.
The composition of described configuration 1000ml substratum is casease hydrolyzate 12.0g-14.0g, yeast soaks powder 10.0g-12.0g, S-WAT 0.20g-0.40g, PXB 0.010g-0.015g, Sulphadiazine Sodium 0.05g-0.10g, ironic citrate 0.60g-0.80g, 4-6 unit of pancreas Suning, agar powder 16.0g-18.0g, adding distil water is to 1000ml.
The composition of described configuration 1000ml substratum is casease hydrolyzate 14.0g-16.0g, yeast soaks powder 8.0g-10.0g, S-WAT 0.40g-0.60g, PXB 0.008g-0.010g, Sulphadiazine Sodium 0.10g-0.15g, ironic citrate 0.40g-0.60g, 6-10 unit of pancreas Suning, agar powder 12.0g-16.0g, adding distil water is to 1000ml.
The composition of described configuration 1000ml substratum is casease hydrolyzate 16.0g-20.0g, yeast soaks powder 6.0g-8.0g, S-WAT 0.60g-0.80g, PXB 0.005g-0.008g, Sulphadiazine Sodium 0.15g-0.20g, ironic citrate 0.20g-0.40g, 10-12 unit of pancreas Suning, agar powder 10.0g-12.0g, adding distil water is to 1000ml.
Described distilled water is distilled water.
Described substratum is by high pressure steam sterilization, and its force value is 103.4kPa.
The final pH value of described substratum is 7.0 ± 0.2 under 25 ℃ of conditions.
Layoutprocedure: under 35-37 ℃ temperature condition, add an amount of distilled water earlier to container, after again casease hydrolyzate, yeast being soaked powder, S-WAT, PXB, Sulphadiazine Sodium, ironic citrate, pancreas Suning, agar powder and adding in the distilled water fully dissolving, be settled to 1000ml with volumetric flask, making its final pH value under 25 ℃ by interpolation acid (HCL) or alkaline matter (NaOH) in solution is 7.0 ± 0.2, at last by 103.4kPa (1.05kg/cm
3) high pressure steam sterilization, need be during packing, and be sub-packed in aseptic plate at Bechtop.
Advantage of the present invention: incubation time obviously shortens than traditional method, and other bacterium with identical cultural characters is suppressed all and does not grow, thereby has realized the effect of fast culture and evaluation clostridium perfringens.
Embodiment:
Under 35-37 ℃ of condition, add an amount of distilled water earlier to container, after again casease hydrolyzate, yeast being soaked powder, S-WAT, PXB, Sulphadiazine Sodium, ironic citrate, pancreas Suning, agar powder and adding in the distilled water fully dissolving, be settled to 1000ml with volumetric flask, making its final pH value under 25 ℃ by interpolation acidity or alkaline matter in solution is 7.0 ± 0.2, at last by 103.4kPa (1.05kg/cm
3) high pressure steam sterilization, need be during packing, and be sub-packed in aseptic plate at Bechtop.
The casease hydrolyzate, yeast soaks powder, and the required nitrogenous source of bacterial growth, carbon source and somatomedin are provided; Agar powder is the vehicle of substratum; PXB, Sulphadiazine Sodium suppress other varied bacteria growing; Pancreas Suning is a positive growth factor; Ironic citrate and S-WAT are colored indicator, and S-WAT also can suppress other Gram-positive bacteria growing.
Cultivated 8-12 hour at 35-37 ℃ with this substratum, the clostridium perfringens well-grown, bacterium colony shows black, and the Gram-positive and the negative bacterium of other reduction sulphite are not grown, thereby realize fast culture and identify clostridium perfringens.
Embodiment 1
A kind of substratum is provided, is used for fast culture and identifies clostridium perfringens.Main component is: casease hydrolyzate 12.0g, yeast soaks powder 12.0g, S-WAT 0.20g, PXB 0.015g, Sulphadiazine Sodium 0.05g, ironic citrate 0.80g, 4 units of pancreas Suning, agar powder 18.0g.The accurate above composition of weighing after fully dissolving with distilled water, is settled to 1000ml with volumetric flask, at 103.4kPa (1.05kg/cm
3) condition under carry out high pressure steam sterilization, it is standby to be sub-packed in aseptic plate, final PH (25 ℃) value is 6.8.Cultivated 8 hours under 35 ℃ of conditions with this substratum, the clostridium perfringens well-grown, bacterium colony shows black, and the Gram-positive and the negative bacterium of other reduction sulphite are not grown, thereby realize fast culture and identify clostridium perfringens.
Embodiment 2
A kind of substratum is provided, is used for fast culture and identifies clostridium perfringens.Main component is: casease hydrolyzate 20.0g, yeast soaks powder 6.0g, S-WAT 0.80g, PXB 0.005g, Sulphadiazine Sodium 0.20g, ironic citrate 0.20g, 12 units of pancreas Suning, agar powder 10.0g.The accurate above composition of weighing after fully dissolving with distilled water, is settled to 1000ml with volumetric flask, at 103.4kPa (1.05kg/cm
3) condition under carry out high pressure steam sterilization, it is standby to be sub-packed in aseptic plate, final PH (25 ℃) value is 7.2.Cultivated 12 hours under 37 ℃ of conditions with this substratum, the clostridium perfringens well-grown, bacterium colony shows black, and the Gram-positive and the negative bacterium of other reduction sulphite are not grown, thereby realize fast culture and identify clostridium perfringens.
Embodiment 3
A kind of substratum is provided, is used for fast culture and identifies clostridium perfringens.Main component is: casease hydrolyzate 14.0g, yeast soaks powder 10.0g, S-WAT 0.40g, PXB 0.01g, Sulphadiazine Sodium 0.10g, ironic citrate 0.60g, 6 units of pancreas Suning, agar powder 16.0g.The accurate above composition of weighing after fully dissolving with distilled water, is settled to 1000ml with volumetric flask, at 103.4kPa (1.05kg/cm
3) condition under carry out high pressure steam sterilization, it is standby to be sub-packed in aseptic plate, final PH (25 ℃) value is 7.0.Cultivated 10 hours under 36 ℃ of conditions with this substratum, the clostridium perfringens well-grown, bacterium colony shows black, and the Gram-positive and the negative bacterium of other reduction sulphite are not grown, thereby realize fast culture and identify clostridium perfringens.
Embodiment 4
A kind of substratum is provided, is used for fast culture and identifies clostridium perfringens.Main component is: casease hydrolyzate 16.0g, yeast soaks powder 8.0g, S-WAT 0.60g, PXB 0.008g, Sulphadiazine Sodium 0.10g, ironic citrate 0.40g, 10 units of pancreas Suning, agar powder 12.0g.The accurate above composition of weighing after fully dissolving with distilled water, is settled to 1000ml with volumetric flask, at 103.4kPa (1.05kg/cm
3) condition under carry out high pressure steam sterilization, it is standby to be sub-packed in aseptic plate, final PH (25 ℃) value is 6.9.Cultivated 9 hours under 36 ℃ of conditions with this substratum, the clostridium perfringens well-grown, bacterium colony shows black, and the Gram-positive and the negative bacterium of other reduction sulphite are not grown, thereby realize fast culture and identify clostridium perfringens.
Embodiment 5
A kind of substratum is provided, is used for fast culture and identifies clostridium perfringens.Main component is: casease hydrolyzate 18.0g, yeast soaks powder 10.0g, S-WAT 0.60g, PXB 0.012g, Sulphadiazine Sodium 0.15g, ironic citrate 0.60g, 10 units of pancreas Suning, agar powder 16.0g.The accurate above composition of weighing after fully dissolving with distilled water, is settled to 1000ml with volumetric flask, at 103.4kPa (1.05kg/cm
3) condition under carry out high pressure steam sterilization, it is standby to be sub-packed in aseptic plate, final PH (25 ℃) value is 7.1.Cultivated 11 hours under 37 ℃ of conditions with this substratum, the clostridium perfringens well-grown, bacterium colony shows black, and the Gram-positive and the negative bacterium of other reduction sulphite are not grown, thereby realize fast culture and identify clostridium perfringens.