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VBNC (viable but noncultivable)-like cultivable lactobacillus
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A technology of lactic acid bacteria and preservation number, applied in the field of cultivable lactic acid bacteria, can solve the problems of difficult stability of biological properties, sudden disappearance, short storage period, etc.
Inactive Publication Date: 2015-04-22
JILIN AGRICULTURAL UNIV
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[0004] The purpose of the present invention is to solve the bacteria in the maintenance period of VBNC, whose biological properties are difficult to exist stably, easily disappear suddenly during the research process, and have a short storage period problem, but provide a kind of unfavorable conditions such as resistance to oligotrophy and low temperature, after entering the VBNC state, the VBNC bacterial mutants that can be cultivated by conventional culture methods, can stably reproduce and inherit, and can be cultivated in a non-cultivable state like live Lactobacillus
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Embodiment 1
[0018] Example 1 Chemical mutagenesis of Lactobacillus plantarum
[0019] Lactobacillus (L) was isolated from naturally fermented yoghurt by the Preventive Veterinary Medicine Laboratory of Jilin Agricultural University, and was identified as Lactobacillus plantarum by 16sRNA ( Lactobacillus plantarum ). Preserved by the Laboratory of Preventive Veterinary Medicine, Jilin Agricultural University.
[0020] 1) Obtaining the culturable Lactobacillus MVR strain in a non-culturable state that seems to be alive.
[0021] Experimental group: take 5×10 8 Add 100μl NTG (1mg / ml) to 1ml of Lactobacillus (L) cell suspension per ml, mix thoroughly, and react at 30°C for 30min, after the reaction, use 0.1M phosphate buffer [pH7.0] To terminate the reaction, the dosage of NTG is appropriate to achieve a lethal rate of 80% to 95%.
[0022] Control group: Lactobacillus (L) suspension not treated with NTG.
[0023] The bacterial solutions of the experimental group and the control group ...
Embodiment 2
[0026] Example 2 Feeding the cultureable Lactobacillus MVR strain in a non-culturable state that seems to be alive, and its influence on weaned piglets
[0027] Grouping of experimental animals The weaned piglets were randomly divided into four groups: 1, 2, 3, and 4. Among them, Group 1 was fed with VBNC Lactobacillus mutant, Group 2 was fed with VBNC Lactobacillus, Group 3 was fed with Lactobacillus, and Group 4 was blank control. Groups 1 to 3 contained 10 per gram of feed 9The standard of 1 CFU of live bacteria was added to the feed, and the feeding was carried out in circles and ad libitum. Feed 3 times a day, the test period is 7d. After raising for 7 days, stop feeding each experimental strain, and still contain 10 per gram of feed. 9 Escherichia coli was added to the standard of 1 CFU viable bacteria count, and the diarrhea situation of each group was observed every day. The next day after the occurrence of diarrhea, continue to feed for 7 days according to the a...
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Abstract
The invention discloses a VBNC (viable but noncultivable)-like cultivable lactobacillus which is the latest researched and discovered novel bacterium existence form, is a VBNC bacteria mutant, has the VBNC bacteria-related gene after entering into the VBNC state, and can be cultured by a normal method without the resuscitation of the resuscitation method of the viable but noncultivable state of the bacterial activity. The VBNC bacteria mutant of the lactobacillus plantarum is preserved at the China General Microbiological Culture Collection Center at 18th Sep 2012. The regret that the VBNC bacteria can not be grown, reproduced and preserved can be remedied; and the lactobacillus is taken as a VBNC bacteria model to research the generation mechanism of the VBNC, the lactobacillus has wide prospect in the aspect that the lactobacillus is taken as a carrier to research the oral vaccine and the like, and the immunologic activity of the VBNC lactobacillus and the mutant of the lactobacillus to the weaned pig can be detected, so that the cultivable lactobacillus lays a foundation for further preparing the novel pig microecologics.
Description
technical field [0001] The present invention relates to a living form of bacteria, specifically cultivable lactobacillus in a non-culturable state that seems to be alive. Background technique [0002] As we all know, the animal microecological live bacteria preparations commonly used at present are mainly developed from various probiotics such as lactic acid bacteria, bacillus, and yeast. Among them, the most widely used and most diverse live bacterial preparations are mainly lactobacillus. However, because Lactobacillus is not heat-resistant, it cannot withstand the destructive effects of high temperature and high pressure in the process of feed granulation and puffing, so many experts strongly suggest that they should be fed directly. However, the most important sign of live bacteria preparation products for direct feeding is the number of viable bacteria, and it is stipulated that the number of viable bacteria in qualified products should reach 10 9 CFU / g, so the number...
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