262 THE PROTOZOA 



get serum in that way or by squeezing the chancre afterward. Mix and make a 

 smear as for blood. When dry examine with the oil immersion objective and the 

 treponemata will be found to stand out as white spirals against a dark background. 

 Treponemata often appear as if bent in the middle. 



Harrison prefers collargol to india ink. One part of collargol is put in a bottle 

 with 19 parts of water and well shaken. This shaking is repeated. One loopful 

 of the suspected serum and one loopful of the collargol suspension are mixed and 

 smeared out and examined as for the india-ink method. 



Cultivation. T. pallidum has been cultivated anaerobically in horse serum by 

 Schereschewsky. The cultures contained other organisms. Muhlens, by growing 

 anaerobically on horse-serum agar (i to 3), claims to have obtained pure cultures. 

 Animal inoculations with this material were negative, however. 



Noguchi has cultivated T. pallidum under strict anaerobic conditions in a medium 

 of ascitic fluid containing a piece of fresh sterile tissue, preferably placenta. The 

 growth is faintly hazy and does not have an offensive odor. Spirochceta micro- 

 dentium shows similar morphology but the cultures have a foul odor. Sp. macro- 

 dentium is similar culturally but differs morphologically. 



Luetin. When cultures of T. pallidum, grown for one or more weeks in ascitic 

 fluid agar and ascitic fluid are ground in a mortar, heated to 6oC. for one hour 

 then, with the final addition of i% trikresol, we have an emulsion called "luetin." 

 This extract produces an allergic reaction on the skin of certain syphilitics (luetin 

 reaction). To carry out the test luetin is introduced intradermally at the insertion 

 of the left deltoid and a control emulsion of agar media injected in the right arm. 

 A negative result shows as an erythema without pain or papule formation. Positive 

 reactions show as papules vesicles or even pustules giving rise to discomfort for 

 several days. Not only do we have papular and pustular type reactions but also 

 torpid ones (taking ten days or more to develop). While the control side usually 

 becomes normal in forty-eight hours yet in latent and tertiary syphilis the control 

 may show almost as marked a reaction. The term "Umstimmung" is applied to 

 this susceptibility to trauma of the skin of those having tertiary syphilis. Some 

 cases of parasyphilitic infections which are negative to the Wassermann test give 

 a positive luetin reaction. Tertiary yaws cases frequently give a positive luetin 

 reaction. See comparison of Wassermann and luetin statistics. 



Noguchi has recently demonstrated T. pallidum in all layers of the cerebral 

 cortex except the outermost one in 12 cases out of 70 cases of general paresis 

 examined. 



Diagnosis. In diagnosis either use the dark ground illuminator or make a thin 

 smear from the sanious oozing after vigorous friction of the chancre with gauze, 

 taking up this blood-stained serum on the end of a slide and smearing the surface 

 of a second slide with the adhering material. It is in most cases more satisfactory 

 to curet the lesion, in this way obtaining material from the areas of the thickened 

 arteries. Fontana's silver staining method is an excellent one. 



In the diagnosis of cerebrospinal syphilis we use, in addition to the Wassermann 

 test of the blood, (i) the Nonne-Apelt reaction in which about i c.c. of a saturated 

 aqueous solution of ammon. sulphate is added to an equal amount of cerebrospinal 

 fluid. If turbidity or rather opalescence appear immediately, or within three 

 minutes, the test is positive. We now use a ring test. (2) The counting of the 

 lymphocytes in the cerebrospinal fluid. A lymphocytosis occurs in cerebrospinal 



