A kind of Clostridium butyricum slzx19-05 and its application
A technology of Clostridium butyricum and bacteria agent, applied in the field of probiotics, can solve the problems of drug resistance gene transmission, environmental pollution, shortage, etc., and achieve the effect of strong sensitivity, high tolerance, and promotion of production
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Embodiment 1
[0056] Example 1 Isolation and Identification of Clostridium butyricum SLZX 19-05
[0057] 1. Isolation of strain SLZX19-05
[0058] Clostridium butyricum (Clostridium butyricum) SLZX 19-05 was isolated from the excrement of Tibetan pig breeding cooperatives in Longzi County, Shannan City, Tibet Autonomous Region. The specific isolation method is as follows:
[0059] 1. Preliminary screening: take 1g of Tibetan pig manure sample, mix it in sterile saline, heat it in a water bath at 80°C for 10 minutes, and dilute it in a gradient with sterile saline. Spread evenly on a 0.04g / L RCM agar plate, culture in an anaerobic incubator at 37°C for 24 hours, randomly select single colonies with differences on the plate according to the colony shape and acid production, number them and inoculate them on the RCM agar plate for marking Line purification culture.
[0060] 2. Re-screening: take 1% volume seed solution (OD=0.8 or so) of the bacteria with acid-producing ability initially iden...
Embodiment 2
[0069] Example 2 Characteristic Detection of Clostridium butyricum SLZX 19-05
[0070] Activation of seed solution: inoculate Clostridium butyricum SLZX 19-05 into sterilized RCM liquid medium, leave it in anaerobic state at 37°C for 16 hours, then take it out for use.
[0071] 1. Determination of growth curve
[0072] Draw a certain amount of Clostridium butyricum bacteria liquid from the seed liquid according to 1% inoculation amount and inoculate it into RCM liquid medium (OD600=0.176), put it into an anaerobic workstation, and culture it statically at 37°C. Use a microplate reader to measure the OD600 value of the fermentation broth every 2 hours, and save the fermentation broth for each period of time to count the viable bacteria.
[0073] The results show( image 3 and Figure 4 ), the Clostridium butyricum SLZX 19-05 strain entered the logarithmic growth phase at 6 h, entered the plateau phase at 12 h, and the maximum number of viable bacteria reached 1.6×10 7 CFU / m...
Embodiment 3
[0084] Example 3 Effect of Clostridium butyricum SLZX 19-05 on mouse intestinal function
[0085] 1. Experimental method
[0086] 1. Select about 12g C57BL / 6J male mice one week after weaning, use drinking water dissolved in 0.5g / L vancomycin, 1g / L neomycin sulfate, 1g / L metronidazole, 1g / L ampicillin , to construct a pseudo-sterile mouse model, select 18 pseudo-sterile mice after 4 weeks, and randomly divide them into three groups (Con group is the control group, fed with sterile normal saline; Low dose group is the low-dose Clostridium butyricum group, Contains 2.1×10 5 CFU / mL Clostridium butyricum normal saline; the High dose group was a high dose Clostridium butyricum group, and the concentration of Clostridium butyricum was 2.1×10 6 CFU / mL saline), 6 mice per group.
[0087] 2. Gastric gavage method: gavage once a day, 200 μL each time, stop gavage after colonization of Clostridium butyricum was detected in the feces on the third day, and collect chyme and tissue sampl...
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