A NOTE UPON THE USE OF ANTIFORMIN IN SPUTUM EXAMINATION, AND ON STAINING METHODS FOR THE DEMONSTRATION OF TUBERCLE BACILLI

AND\ls ON\t STAEVI,NG,66. METHODS FOR TIIEC DE.11ONST]wRA4treated with antiformlii NTION OF lETHBERCLE BTACSILHDL J and further investigate TION OFTUBE A ILLL has been relatively 'largE By G. H. K. MACALISTER, M.D., D.P.H. , thlat statistics based ASSISTANT BACTERIOLOGIST, LISTER INSTITUTE. sufficient value to justif3 The nuimber of sputa SIN-CE the routine examination of sputum as a mleans of thirteen months has b liagnosis for tuibercle became a universal practice, it has 622 were foLnd to be pos beeni recognized that diagnosis might be rendered more mens, which gave negat precise by some satisfactory miiethod of rendering the were treated with anti sputuni homogeneous, and dissolving the non-tubercutlous showed thc presence o elemi1ents therein contained. For this purpose suclh repercentage of cases fou ag,ents as caustic alkallis, hydrogen peroxide, etc., were was thus 27.36, and ti tried w^itlh a certain measure of success. The procedure 27.76. The cases foun of Ellermann and Erlandsen (1908), which consisted ill treated with antiformir autodigestion of the sputum in the presence of dilute total number of sputa in sodiuna carbonate, and subsequent treatment witll dilute those found negative or sodium lhydrate, was shown by careftul workers (JRrgensen, 0.54 may be described m 1910) to give results superior to those . obtained with percentage antiformin, but, nevertheless, tlle introdutction of this latter reagent must be regarded as the chief recent Co0mparison i advance in this branclh of study. Antiformiiin is a Other observers have proprietary substance whliclh is made up of eciual of improvement with parts of caustic soda 15 per cent. , and liquor sodae sliown by our figures. chlorinatae. It was first used in breweries for c(leausing thle metlhod, Meyer (19 vats and for general disinfection. It is a very powerful examination of 114 sa solvent of organic substances, rapidly causing the found positive on dire disintegration and solution of suclh materials as pus, mucus, 99 samples were treat connective tissues, faeces, and hair. Waxes and waxv investigations revealed substances alone resist its action. -Uhlenhuth and 14 cases. Other contri Xylander (1909) studied its solvent and its germnicidal subject, too numerous t powers, alnd published the description of a method for its ment percentages rangin applicationi to the investigation of tuberculous material. has recently miiade a c They found tlhat, whereas otlher bacteria dissolved in the principal modifications c reagent as rapidly as sugar in water, ttubercle bacilli obtained such improvex retained tlleir vitality in a 20 per cent. solution for twelve He recoimmends the us to twenty-four houlrs, and continued to exhibit tlheir that the examination be characteristic fornm and staining reactions for eight days. material has become hoi Ulhlenhuith made use of a 20 or 25 per cent. dilution of of bacilli, wheu very f4 antifomrnin. This was added to spututm in equal bulk. method of Lorenz the 1 The mixture was well shaken and incubated at 370C. for sLx involvinlg the ulse of ] to twelve hours. Then after centrifugalization; the deposit reports unfavourably, be was examined for tubercle bacilli in the ordinary way. lhave a detrimenital eff Lorenz (1911) used the same strength of antiformin, but acid-fast bacilli. lheated the mixture over a flame, thus rendering the In order to reconcile t sputum more rapidly lhomogeneous. Haserodt (1909) slhoolk Institute with those qu( uip tlle sputuLm and antiformin mixtture, after it lhad sider tlle procedure tha become lhomogeneous, with ligroin, and placed the mixture in the first instance t in a water bath at60° C. The lighter fluid rising to the sputum for examinatiol top, carried with it the tubercle bacilli, wlicll collecte(d at examined, if need be, for the zone of juncetion between the two layers of fluid. For cases tubercle bacilli, if this method no centrifug,e is required. first lhalf-minute, but in Whlen rapid concentration of tubercle bacilli in sputum detected after three or R is required', Hammerl's method (1909) is satisfactory. One two of the nine cases in part of sputum is shaken up with two parts of a 1 per cent. after antiformin treatm solution of caustic potash in strong ammonia. The filimi showed the presence mrixtture is incubated for half an lhour, being shaken uip prepared from material I flomi time to time. Tlheni, after the addition df one part of fornmiin are examninied wi acetone, to lower the specific gravity of thc liquid, it is filmli. Here bacilli, if prE centrifugalized and the sediment examined. Tlhis metlhod lnumbers, but in one in I have tried in a nuLimber of cases with good results. By after a protracted searcel this miieans, lowever, tubercle bacilli are rapidly killed, and ing to note tlle worlk tlierefore the method cannot be used for the purpose of twenty-four lhours' sput obtaining pture cultures of tuberele. Antiformini, which patients, and foullid in does not kill tubercle bacilli if used in sufficient dilution, is two, four, or six bacilli. well adapted for such cultural pul-poses. No paper previously In the routine work of tlle Lister Institute, the sample with such a large numbi of sptutum is lmlixed with ani equal quantity of a 30per cent. and the results of tliese solution of antiformin, and the mnixtare is incubated overthe greater the care expE night at 370 C. After centrifuigalization, the supernatanit direct film, the les, vil fllid is poured away and replaced by an ecqual bulk of use of antiformin. Thlis iioriial saline soltution. Tinis is slhaken up witli the of tllc antiformin metho deposit, and the mixtire again centrifugalized. Films as low as 0.54 involve fromii tlc deposit tlhus washed adlhere better to the slide precision, whliclh is of i tlhan tlhose prepared after one celntrifugalization only. specimens examined is L TIhe anitifon-imin is diluted for use with distilled water. It was suggested by B3itzke (1910) that tile use of tal) water for this purpose miight introdiuce a fallacy, owing to By means of treating the occasiolnal prescence in suclh water of acid-fast bacilli. samples of sputtum found Organii ni of this type have been found on water taps at | examination, it was pos thle Lister Instituite. I of tuberele bacilli in 9 ea r THE BTITIER 4 MEDICAL JOURNAL 41