Streptococcus suis 7-type high-density fermentation medium and special strain

A high-density fermentation and fermentation medium technology, which is applied in the screening of special strains and the field of high-density fermentation medium, can solve the problems of unsuitable for large-scale enrichment, high cost, and slow growth of bacteria

Active Publication Date: 2012-12-05
HUAZHONG AGRICULTURAL UNIVERSITY +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although this medium is easy to use, it has the disadvantages of slow bacterial growth and high cost during the cultivation process. 2 billion, not suitable for large-scale enrichment

Method used

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  • Streptococcus suis 7-type high-density fermentation medium and special strain
  • Streptococcus suis 7-type high-density fermentation medium and special strain
  • Streptococcus suis 7-type high-density fermentation medium and special strain

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Embodiment 1: Isolation and identification of Streptococcus suis 7-YZ

[0054] 1. Isolation of Streptococcus suis 7-YZ strain

[0055] The disease materials to be separated - the disease materials of various tissues were obtained by the applicant in August 2005 from the disease materials submitted for inspection by a pig farm in Yongzhou City, Hunan Province. The specific separation method is as follows: aseptically collect heart blood, lung tissue, spleen tissue, and synovial fluid, inoculate them on a TSA plate, and observe after culturing at 37°C for 12-24 hours. Pick small colonies with a diameter of 0.1mm-1.0mm, off-white, translucent, smooth, round, and with neat edges, and inoculate them in TSB liquid medium for overnight culture on a shaker at 37°C. Purely cultured bacteria were subjected to Gram staining for microscopic examination. Then observed with an optical microscope, Gram staining was positive, double or short streptococcus, and it was subcultured and ...

Embodiment 2

[0077] Embodiment 2: Fermentation medium component optimization test:

[0078] Taking the number of live bacteria (cfu) as the screening index, using a single factor experiment, first from many carbon sources (glucose, lactose, sucrose, maltose, glycerin, citric acid); nitrogen sources (soybean peptone, yeast powder, peptone, beef extract, pancreas peptone); inorganic salts and organic acid factors (Na 2 PO4, K 2 HPO4, MgSO 4 , CaCl 2 , (NH 4 ) 2 SO 4, ammonium acetate, and sodium citrate to screen out some important factor screening designs, using reasonable experimental design, considering the orthogonal effects of important carbon sources, nitrogen sources, inorganic salts and organic acid factors, etc., for Find the steepest climbing test in the optimization area near the maximum value, and use the Central Composite Design to optimize the maximum value of the response surface to fit the functional relationship between the factors and the response value to obtain the ...

Embodiment 3

[0084] Embodiment 3: the shaking flask fermentation culture of Streptococcus suis 7-YZ

[0085] (1) Preparation of first-class seeds: Streptococcus suis 7-YZ with the preservation number CCTCC NO: M2011160 was streaked on a TSA plate, cultured at 37°C for 16-24 hours, and then picked out with a diameter of 0.1mm-1.0mm, Small off-white, translucent, smooth-surfaced, round, and neat-edged colonies were inoculated on several TSA plates, cultured at 37°C for 16 hours, and first-grade seeds were obtained. Or store it at 2-8°C for later use (it should not exceed 5 days, and it should not exceed 5 generations on the culture medium).

[0086] (2) Preparation of secondary seed liquid: use an inoculation loop to take a single colony of the primary seed of step (1), inoculate it in a commercially available TSB liquid medium, place it on a shaker at 37°C at 170rpm / min, and cultivate it for 6 hours , so that OD 600 Reached 0.713, obtained the secondary seed bacterial liquid.

[0087] (3...

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Abstract

The invention belongs to the technical field of agricultural microbes and specifically to isolation and screening as well as fermentable cultivation of a streptococcus suis 7-type strain. A streptococcus suis 7-YZ strain with a preservation number of CCTCC NO: M2011160 is obtained through separation. The fermentation technology comprises the steps of: 1) taking streptococcus suis 7-YZ strain cant seeds for streak cultivation on an TSA culture medium flat panel to obtain first-class seeds; 2) grafting first-class seed single colony into a TSB (Trypticase Soy Broth) culture medium shake flask to enable the OD600 to be 0.7-0.9 so as to obtain second-class seeds; 3) grafting the second-class seeds to the shake flask for fermentable cultivation so as to obtain a streptococcus suis 7-YZ bacteria liquid. According to the streptococcus suis 7-type high-density fermentation medium and the special strain, disclosed by the invention, growth and metabolism of streptococcus suis 7-YZ strain are facilitated, fast growth speed of the thallus is achieved, relatively high bacteria amount can be obtained, 6-7 billions of the streptococcus suis 7-YZ viable counts are cultured. The production cost is decreased by 30-50% when being compared with that of a commercial TSB culture medium.

Description

technical field [0001] The invention belongs to the technical field of agricultural microorganisms, and specifically relates to a high-density fermentation medium suitable for Streptococcus suis type 7 and its application. The invention also includes the screening of special bacterial strains suitable for the above-mentioned medium. Background technique [0002] S. suis infection has become one of the major problems in the swine industry worldwide. It can cause meningitis, arthritis, endocarditis, pneumonia and sepsis in pigs, and can also cause human death. At present, 33 serotypes have been found according to the different capsular antigens, among which the pathogenic S. suis serotypes that can cause human and animal diseases are mainly type 1 (SS1), type 2 (SS2), and type 7 ( SS7) and type 9 (SS9) [Wisselink et al., 2002]. Streptococcus suis type 2 is the most prevalent serotype. In addition to type 2 strains, type 7 strains are also one of the frequently isolated serot...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12R1/46
Inventor 金梅林陈关平陈守文徐高原康超王杨波陈波何伟韩进曹毅陈焕春
Owner HUAZHONG AGRICULTURAL UNIVERSITY
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