枯草芽孢杆菌作为新的益生菌演讲稿课件.ppt
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1、枯草芽孢杆菌作为新枯草芽孢杆菌作为新的益生菌演讲稿的益生菌演讲稿CONTENTS01020304INTRODUCTIONMATERIALS AND METHODSRESULTS AND DISCUSSIONCONCLUSIONINTRODUCTIONProbiotics:live microorganismspositive effect on diverse functions of an organismprevent the invasion of various pathogens Experiments with mice proved that B.subtilis spores
2、were able to germinate,and the cells could proliferate and form the spores again in intestines of animals Autochthonous lactic acid bacteria of the genera Lactobacillus and Bifidobacterium are usually used as probiotics;gram positive spore-forming bacteria of thegenus Bacillus are used as probiotics
3、 as well Studies have revealed that in addition to the resistance of the probiotic Bacillus strains to bile acids,they were capable of immune stimulation in case of gastrointestinal disorders.Ability to synthesize antibiotics,bacteriocins,cyclic lipopeptides,and lytic enzymes with antimicrobial acti
4、vity provides probiotic activity for bacteria of the genus Bacillus.Being probiotics,the strains of B.subtilis and B.coagulans were shown to have a growth-stimulating and prophylactic effect on broiler chickens1INTRODUCTION标题The goal of the present work was to characterize the properties of these ne
5、w strains and to assess their potential use as probiotics.01INTRODUCTIONMATERIALS AND METHODSThe subjects of the study were B.subtilis strains GM2 and GM5 with high antimicrobial activity(Mardanova et al.,2017).Opportunistic coliform bacteria,Salmonella enterica serotype Typhimurium ATCC14028s,Klebs
6、iella oxytoca,and Escherichia coli,as well as micromycetes Fusarium avenaceum,F.oxysporum,F.redolens,and F.solani,were used as test cultures.02MATERIALS AND METHODSNutrient media and cultivation conditions.The following media were used for cultivation.These were:(1)LB(LuriaBertoni)medium(g/L):trypto
7、ne,10.0;yeast extract,5.0;NaCl,5.0;(2)LA medium(g/L):tryptone,10.0;yeast extract,5.0;NaCl,5.0;agar,20.0;(3)medium no.1(g/L):maltose,20.0;peptone,10.0;CaCl2,1.0;(4)medium no.2(g/L):soybean meal,30.0;NaNO3,3.0;K2HPO4,1.0;MgSO4,0.2;KCl,0.2;(5)medium no.3(g/L):corn extract,20.0;lactose,10.0;(6)deoxychol
8、ate citrate agar for salmonella cultivation.Bacteria were cultivated in a thermostat at 37C and using an IKAKS 4000 incubator shaker(Germany)at 37C and rotation speed of 200 rpm.Optical density of the culture was measured using a Bio-Rad spectrophotometer(United States)at a wavelength of 590 nm.02MA
9、TERIALS AND METHODSThe dynamics of bacterial growth and spore formation were studied on the LB medium and the medium no.1.Bacteria were cultivated for 48 h using the incubator shaker(37C,200 rpm).Bacterial culture samples were collected every 2 h in order to measure optical density(at the wavelength
10、 of 590 nm)using a spectrophotometer and to determine the extracellular proteolytic activity.The numbers of spores formed were counted in a Goryaev chamber(Optical Market,Ukraine),starting after 10 h of cultivation of bacteria.The bacteria were examined under a MICROS AUSTRIA MC 300 microscope(Austr
11、ia).02MATERIALS AND METHODSAbility to grow at various values of ambient pH wasstudied in 250-mL Erlenmeyer flasks containing 50 mL of the LB medium(pH 210).Bacteria were cultivated using the incubator shaker(37C,200 rpm,aeration).Optical density of the cultures was determined after 24 h of growth.Re
12、sistance of the spores to pH and spore capacityfor in vitro germination were studied according to themethod described(Haller et al.,2001).Spores of bacilli were obtained by heating a 2-day-old bacterial culture for 90 min at 60C to eliminate the vegetative cells.The initial concentration of the spor
13、es was 4 107 CFU/mL.The spore suspension in LB medium was incubated at various pH values at 40C.The spores were incubated at pH 5.0 for 60 min and concentrated by centrifugation(6000 rpm,10 min).The supernatant was removed;the spore pellet was transferred into the same volume of the fresh LB medium(
14、pH 3.0)and incubated for 90 min.An aliquot of the suspension(0.1 mL)was taken and heated for 90 min at 60C.A series of dilutions was prepared and plated to form a lawn on the LA medium for the subsequent enumeration of CFU/mL.The remaining spore suspension was concentrated by centrifugation,transfer
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