Preliminary Research And Application Of A Simple Anaerobic Culture Device

Anaerobic bacteria refers to a type of microorganism that grows under anaerobic conditions or under conditions of low redox potential.The main metabolic mode is anaerobic fermentation.Due to a series of physiological characteristics of anaerobic bacteria,such as the lack of superoxide hydrogen dismutase,catalase and cytochrome oxidase in the cells,Therefore,it can only be achieved under strictly anaerobic conditions and in a lower redox potential environment.This has brought great difficulties to the isolation,cultivation,research,and application of anaerobic bacteria.At present,the basic methods of anaerobic bacteria cultivation in the laboratory are Hungate tube rolling technology,anaerobic tank technology,and anaerobic glove box technology,in addition to media deoxidization technology,steel fiber deoxidization technology,gas bag anaerobic system and anaerobic culture dish technology.Because of the high requirement of these anaerobic culture methods and technical equipment,the high cost,the complicated operation,and the general laboratory do not have the conditions,thus the extensive research work of anaerobic bacteria is limited.In anaerobic bacteria,lactic acid bacteria are fermentative carbohydrates,the main products of which are lactic acid,a class of non-spore,gram-free bacteria staining positive bacteria,they are mainly heterotrophic anaerobic metabolism.Lactobacillus is the main representative genus of lactic acid bacteria and is widely found in the intestines of livestock and poultry.A lot of studies have shown that Lactobacillus can effectively regulate the normal flora in the gastrointestinal tract of the animal body and maintain the micro-ecological balance of the intestinal tract.However,due to the limitation of anaerobic equipment,there are still individuals and small research teams in China who do not have stable and reliable anaerobic methods and equipment to cultivate and study Lactobacillus.In this study,a simple anaerobic culture device was designed.The device consisted of the anaerobic proliferation tube,anaerobic dilution tube and anaerobic culture box.During the test,we used the anaerobic indicator to test its anaerobic effect.Our anaerobic method was validated by culturing obligate anaerobic bacteria-Bifidobacteria.Lactobacillus was isolated and cultured from the intestinal contents of healthy chickens by using TPY lactobacillus culture medium,and the morphology and culture characteristics of the isolates were observed.Molecular biological methods were used to identify 25 strains of Lactobacillus and one obligate anaerobic bacteria-Veillonella Magna.Among them,ten strains of Lactobacillus reuteri are numbered L-1,L-3,L-6,D-1,Y-3,MY-2,JY-3,MCPY-2,GYM-1,GYM-2;three strains of Lactobacillus oris are numbered D-6,D-7,D-5,one strain of Lactobacillus caviae,numbered Y-1;three strains of Lactobacillus ingluviei,numbered H-2,H-3,JY-4;two strains of Lactobacillus crispatus,numbered D-2,H-4;one strain of Lactobacillus brevis,numbered SY-1;one strain of Lactobacillus pentosus numbered SY-3;one strain of Lactobacillus kitasatonis,numbered GYC-2,one strain of Lactobacillus johnsonii,numbered GYC-1;two strains of Lactobacillus gallinarum,numbered HC-1,HC-2.This study designed a simple anaerobic culture device,which isolated and cultured Lactobacillus and achieved good results.The method is low cost,convenient,effective,stable and reliable.Basically,it can meet the research of anaerobic bacteria by individual and small research teams and has certain promotion value.