April 23, 1887.] THE DETECTION OF THE GONOCOCCUS. 455 A NEW COLOR-TEST (ROUX’S) FOR THE DETECTION OF. THE GONOCOCCUS; WITH REMARKS ON ITS PRACTICAL IMPORTANCE. BY EDMUND C. WENDT, M.D., OF NEW TOWC For the past three years the relation of Neisser’s gonococci to gonorrhoea has been engaging my attention. My studies were pursued with the view mainly of adding to our clinical knowledge of the subject, and were carried on conjointly with Dr. Charles W. Allen, of New York. I was early struck with the difficulty, if not the impossibility, of differ- entiating the microbes of blennorrhcea from similar diplococci. Like other observers, I was unable to confirm Neisser’s statement regarding the characteristic appearance of the specific micrococci of gonorrhoea. And, indeed, it is now generally admitted by those most competent to judge, that the gonococcus is not sufficiently well characterized by its morpho- logical attributes, to admit of our establishing its identity. Again, while it is quite true that, in most cases, the peculiar intracellular grouping of the gono- cocci will render a diagnosis highly probable, never- theless, I cannot allow to go unchallenged the claim of Bumm, to the effect that this constitutes a truly pathognomonic sign. Among about one thousand specimens, of various secretions examined, I have several times observed this alleged pathognomonic arrangement of microbes within the pus-corpuscles, although gonorrhoea was quite out of the question. It was not until I had become thoroughly ac- quainted with Roux’s color-test for gonococci, that I felt confident that blennorrhoea could be diagnosti- cated with certainty, by the aid of the microscope. The method of Roux was made public at a meeting of the Acad6mie des Sciences, held November 8, 456 THE DETECTION OF THE GONOCOCCUS. [Medical News 1886. After a considerable number of experiments, I am able to state my belief that it is a reliable one. The method itself is based on the peculiarity of gono- cocci to hold aniline colors only very loosely, and to become speedily decolorized when subjected to the action of ordinary reagents for that purpose. Although Bumm has found and described no less than five different kinds of diplococci,1 it does not appear that any but the true gonococci retain aniline colors so poorly as to almost immediately fade from view in the presence of decolorizing fluids. On the con- trary, by employing the method of Gram, the ordinary bacteria, as is well known, appear deeply stained. But in a given specimen stained after the manner of Gram, all gonococci invariably disappear, even more quickly than do the anatomical elements thereof. It appears that Gram’s solution of iodine and iodide of potash, so far from fixing the aniline color upon gonococci, as it does upon the other schizo- mycetes, causes them to give up the stain previously imbibed. I will now describe, step by step, the method of procedure which I have found to yield the best results, and which differs in no essential respects at least, from that advocated by Roux, to whom is due the credit of having first applied it for purposes of differential diagnosis. The material to be examined is spread in a thin film upon a glass slide by draw- ing a second slide over it. It is then allowed to dry at the ordinary temperature of the room.2 The specimen is next stained by one or two drops of a saturated solution of methyl-violet in aniline water. Twenty to thirty seconds are amply sufficient to accomplish this purpose. After washing in distilled water, and placing a cover glass on the slide, it is ready for the first examination. The gonococci are found deeply stained, precisely like all the other microorganisms present in the specimen. The cover- slip is now removed, Gram’s solution of iodine in iodide of potassium (iodine i part, iodide of potas- sium 2 parts, distilled water 300 parts) is poured upon the slide and allowed to act for three minutes. After again washing in water, absolute alcohol is poured over the specimen and quickly removed by a gentle stream of water. The cover-slip is now replaced and the specimen submitted to a second examina- tion. This procedure brings out ordinary bacteria in so strikingly beautiful a manner, that it has been called by Friedlander “nearly ideal.” But while this is true of microbes in general, the gonococci are found to have disappeared. In typical gonorrhoeal pus from the male urethra the gonococci exist in such abundance as to over- shadow by their presence all other microorgan- isms. While the intracellular grouping of the microbes is certainly characteristic, that sign alone should not be relied upon in making a positive diag- nosis. But if, in addition, the bacteria disappear on the application of the above method, it is safe to assume the presence of blennorrhoea. If, on the Fig. i. Usual appearance of the discharge in acute gonorrhoea. A. Intra- cellular group of gonococci (pathognomonic according to Bumm). B. Extracellular group of gonococci. C. Urethral epithelium containing gonococci. Schematic. other hand, these microorganisms persist, the infer- ence is justifiable that they are not gonococci, even though they should occur in groups within the cells. Fig. 2. Usual appearance of discharge in “nearly cured’’ gonorrhoea. Schematic. I have not found it necessary to employ eosine as a contrast-stain, as recommended by Roux. But there is no harm in its employment, and it facili- tates focussing in those cases where only gonococci exist in a specimen. The eosine always causes the anatomical elements to reappear with a faint rosy hue, the gonococci also assuming a very faint color. The other bacteria remain purple, and thus a double coloration appears. But it is necessary to have some experience with the use of eosine, or the speci- men will be spoiled. I employ an alcoholic solution, 1 These diplococci were obtained from various sources, such as ordinary atmospheric air, the preputial smegma, the discharge in acute vaginitis of children, etc. They are all accurately described in the second edition of Bumm’s work, entitled “ Der Mikro- Orgamsmus der Gonorrhoischen Schleimhaut Erkrankungen, Gonococcus Neisser,” Wiesbaden, 1877. 2 I have come to discard the use of heat for drying specimens of gonococci, since their shape is better preserved without passing the glass-slide through the flame. April 23, 1S87.] CASE OF IMPERFORATE HYMEN. 457 and allow it to act about one minute. On the whole, I do not regard it as necessary for purposes of diagnosis. If the experience of others will be found to sub- stantiate, as Dr. Allen’s and my own has done, the claim to absolute accuracy made for his method by Roux, then I doubt not a decided step in advance will have been taken. Those familiar with bacteri- ological methods know only too well how difficult it is to obtain the supreme proof of the etiological significance of the gonococcus by the implantation of pure cultures. Thus so able and competent an observer as Sternberg was compelled recently to ad- mit (The Medical News, February 26, 1887) that he had mistaken a non-pathogenic culture of diplo- cocci for one of the true gonococcus, and that, therefore, the negative outcome of his inoculation experiments was devoid of significance.1 Bumm himself only succeeded after many futile efforts, and according to him the gonococcus grows well, outside of the body, only upon human blood- serum. Moreover, in our country it would be difficult to find healthy men or women who would submit to having their sexual organs tampered with to the extent of artificially producing, or attempting to produce, a typical gonorrhoea. In Germany there appears to be no lack of “human material ” for the experiments of the enterprising mycologist. As gonorrhoea cannot be produced in animals, we must either depend upon microscopical evidence for purposes of diagnosis, or go without. It is for these reasons that the new color-test described above will be welcomed by the practitioner as placing within his reach a ready means of deciding positively as to the true nature of a discharge. A word of caution is perhaps necessary here. Granting the absolute reliability of Roux’s method as applied to intracellular groups of diplococci, it does not follow that the presence, in a suspected discharge, of the microbes of gonorrhoea must result in a communication of disease under favoring conditions. It does prove the infective capacity of a secretion just as positively as the absence of the gonococci proves the opposite. But that is all. It is a well-known clinical fact that exposure to infec- tion does not always result in an attack of the particular disease in question. This is eminently true of gonorrhoea. I have observed cases of mar- ried men whose urethral discharges always showed gonococci, but whose wives remained healthy and gave birth to children showing no sign of ophthalmic blennorrhoea. But it is also true that men with a minimum of discharge containing gonococci have communicated the disease to others. I have never known a case in which infection resulted in the absence of gonococci. And again I have seen a few cases in which all the clinical symptoms of gonorrhoea were present, but in which the abundant creamy discharge contained no gonorrhoeal microbes. In these cases recovery resulted in from two to six days, by letting things alone and ordering a placebo. These are doubtless the cases that appear in literature as “cured” by some “original” or novel plan of treatment. A systematic microscopic examination of all suspected secretions will soon dispel all such illusions. True gonorrhoea is never “cured” in less than four weeks. In many cases the process is of much longer duration. At least this has been my experience with the disease. But true gonorrhoea is not a benign urethral catarrh, it is a virulent inflam- mation due to bacterial invasion. This should be generally recognized by the profession, for there still prevails in our ranks much looseness of con- ception regarding the true nature of the disease, coupled ‘with a disposition to shirk the responsibility which modern methods of bacterioscopy have entailed upon every earnest, honest, and well-meaning prac- titioner. 712 Madison Avenue. 1 See Sternberg's articles in The Medical News, of January 20, 1883, and October 18, 1884.