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Bacillus subtilis peptidoglycan extraction and the impact on immune function

Author: ShengQin
Tutor: PanKangCheng
School: Sichuan Agricultural University
Course: Preventive Veterinary Medicine
Keywords: Bacillus subtilis Peptidoglycan Extract Mice Immune function
CLC: S852.6
Type: Master's thesis
Year: 2011
Downloads: 123
Quote: 1
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Abstract


Objective: This study Pab02 Bacillus subtilis cell wall peptidoglycan separation and extraction and analysis of physical and chemical properties, different concentrations of peptidoglycan on immune function, as probiotics, immunostimulants development and application of theoretical basis. Methods: (1) using trichloroacetic acid Pab02 Bacillus subtilis cell wall peptidoglycan separation and extraction. Analysis of Bacillus subtilis peptidoglycan amino acids; using Bradford assay the protein content; phenol - sulfuric acid method for the determination of its polysaccharide content; using vanillin assay was total lipid content. (2) of Bacillus subtilis peptidoglycan on immune function. Animal groups: 5-week-old Kunming mice 208 were randomly divided into four groups. 0.9% in the control group were injected with sterile saline 0.5 ml / only; low-dose group were injected 0.5 mg / ml solution of Bacillus subtilis peptidoglycan, 0.5 ml / only; middle dose group were injected 1 mg / ml peptide of Bacillus subtilis polysaccharide solution was 0.5 ml / only; high dose group were injected 2 mg / ml solution of Bacillus subtilis peptidoglycan 0.5 ml / only. Detection of the immune mice MEASURES: ① detection thymus, spleen absolute weight, index and delayed-type hypersensitivity. ② using spectrophotometer serum lysozyme, antibacterial activity, serum hemolysin and splenic antibody-producing cells. ③ measured using flow cytometry of peripheral blood and spleen T lymphocyte subsets. ④ using carbon clearance method for the determination of macrophage phagocytosis. ⑤ MTT determination of lymphocyte proliferation stimulation index. Results: (1) Extraction of Bacillus subtilis cell wall peptidoglycan Pab02 yield 11.26%. Amino acid composition analysis shows that the extracted sample peptidoglycan amino acid composition and amino acid composition of Bacillus subtilis cell line. Chemical composition analysis showed that the protein content of peptidoglycan was 53.55%, total sugar content of 32.98%, total lipid was 2.91%. (2) different concentrations of Bacillus subtilis peptidoglycan various immune parameters in mice were improved functions: ① peptidoglycan group absolutely immune organ weight and index were higher, and showed significant dose-effect relationship. Delayed-type hypersensitivity showed footpad thickness of peptidoglycan group increased significantly compared with the control group. Description peptidoglycan can promote the growth and development of immune organs, enhance the body immunity against specific antigens. ② After injection of different concentrations of peptidoglycan serum lysozyme, antibacterial activity, serum hemolysin and splenic antibody-producing cells than the control group, and showed significant dose-effect relationship. Tip peptidoglycan can effectively improve the body's specific and nonspecific immunity. ③ peripheral blood and splenic T lymphocyte subsets show peptidoglycan group CD3 CD4, CD3 CD8, CD3 CD4 / CD3 CD8 were higher, and with peptidoglycan were positively correlated, suggesting that peptidoglycan can improve the body T lymphocyte subsets content, and thus higher on the body to maintain immunity has a good effect. ④ peptidoglycan group carbon clearance rate was significantly higher, and with the increasing trend of peptidoglycan concentration. Description peptidoglycan mice with increased monocyte-macrophage phagocytic function, thereby enhancing the body's non-specific immune function. ⑤ Bacillus peptidoglycan can significantly increase lymphocyte proliferation and stimulation index showed significant dose-effect relationship. Visible peptidoglycan can promote immune cell growth, thus contributing to the development of the immune system, improve immune function.

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CLC: > Agricultural Sciences > Livestock, animal medicine,hunting,silkworm,bee > Animal Medicine ( Veterinary Medicine) > Basic Veterinary Science > Animal Microbiology ( Veterinary Microbiology, )
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