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• W2364133246 abstract "Both in clinical bacteriological work and for the students' practice the culturing methodsfor clostridia used at present are often not satisfactory.We therefore made efforts to modifycertain technical procedures particularly in regard to those used in the isolation and identificationof this of bacteria.After preliminary trials,we found certain useful modifications,which areoffered here for consideration. 1.In the literature,there are various methods for obtaining anaerobic enviroment,neces-sary for the isolation of species of Clostridia.The anaerobic Petri dish technic advocated byWang I-min has been found simple and practical.In his original paper Wang used 1.0 gm.ofpyrogallic acid and 1.0 cc.of a 10% solution of sodium hydroxide to produce the requiredanaerobic space.As to the quantities of the chemicals needed,there was,however no uniforminformation.Besides this,it was found that in the anaerobic spaces so produced the cultureswere not so satisfactory as expected. For these reasons,expeximent of the anaerobic Petri dish method with Special attention forstudying the optimal quantities of chemicals and moisture were carried out.According to ourexperimental results,0.4 gm.of the pyrogallic acid and 1.0 cc.of a 6% of sodium hydroxideadded to the space enclosed in the bottom dish,which contained 12 cc.of a pH 7.6 3% liverdigest agar,and sealed on a glass plate,produced an anaerobic enviroment,which wassufficient for a good growth of the Clostridium tetani, Under such a condition,there wasalso produced,the least amount of moisture. The original paper suggested the use of gauze to absorb the above chemicals,but it hasbeen found that in so doing the presence of the negative pressure in the dish often caused,during the opening of the dish,the formation splashing droplets,which frequently spoiled theculture.In our experiments,small Petri dishes of 6 cm.diameter were therefore used to holdthe solution and this did not only prevent the formation of droplets,but also keep the set upclean. 2.In the clinical specimens obtained from infected wounds,there may be the presence oforganisms with the nature of spreading growth,such as Proteus vulgaries and Clostridiumtetani.This kind of growth is very unfavorable for the isolation of a pure culture.' Toovercome this difficulty,we tried to incorporate solution of boric acid or chloral hydrate atvarious concentrations into the pH 7.63% liver digest agar.It was found that a final dilutionof 0.1% boric acid or 0.05% chloral hydrate gave the most favorable inhibitory action. Between these two substances,the former was found more preferable,because it insures not only abetter growth of the Clostridium tetani,but also a good growth for the other species of thisgenus. 3.Sugar fermentation and protein liquification are two biological tests often used for theidentification of the species of Clostridia.Smith recommeded a high colmn of 0.15% brain andheart infusion broth agar with 1% tryptone as the medium base of the sugar test and an 8%egg powder in brain and heart broth for protein liqufication.We considered that this mediumbase was rather costful and was not a medium base available in most of the small laboratories.Besides,the cloudiness of the egg powder in brain and heart infusion broth was a disadvantagein the detection of the culture.By our experimentation,it was found that a pH 7.4 of 0.2%peptic blood digest agar in high columns were a good substitute for the medium base of thesugar test,because it was observed that in such a medium Cl.tetani,Cl.welchii,Cl.septicum,Cl.sporogenus,and Cl.histolyticum grew satisfactoryly. We tried to put a small cube of coagulated egg white or human serum into each of the highcolumn of the 0.2% peptic digest agar as a substitute for the Smith's egg in brain and heartinfusion broth.After autoclaving,we innoculated the 5 species of Clostridia as mentionedabove and it was observed,within 72 hours of incubation,that the species which possess theactivities of protein liqufication,exhibited the ability of liqufying the protein cubes. Smith used the solution of bromthymol blue on a glass plate to detect the pH changes inthe fermentation tubes.In our opinion,this indicator was somewhat too sensitive,owingto the fact that some species of Clostridia would produce certain substances such as hydrogensulfide which might render the medium slightly acidic,especially in the early stage of incubation.This slight shifting of acidity might be mistaken for the splitting of the sugars.By trial wefinally found that a good substitute for the bromthymol blue is a strip of filter paper permeatedwith bromcresel purple.This indicator paper was made by wetting an ordinary filter paperwith a 1.6% bromcresol purple which had been diluted 50 times with 95% ethanol and driedin the incubator.We inoculated the 5 species of Clostridia into the fermentation tubesconsisted of 7 kinds of sugars,and dipped loopful of the contents of the tubes separately on ourindicator paper every day.In so doing,it enabled us to determine definitly the pH changesof the 5 known organisms within 72 hours during incubation. We adapted these modified technics to examine a few specimens from clinic and soil withsatisfactory results." @default.
• W2364133246 created "2016-06-24" @default.
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• W2364133246 date "1957-01-01" @default.
• W2364133246 modified "2023-09-23" @default.
• W2364133246 title "SOME SUGGESTIONS ON THE CULTURING TECHNICS FOR CLOSTRIDIA" @default.
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