AbstractHydrolysis of the azlactones (I) followed by treatment with the thiosemicarbazide (II) yields the thioxo‐1,2,4‐triazinones (III).

The present review covers fluoroquinolone usage in chronic otitis suppurativa (COS) in case of chronic otitis media, cholesteatoma, radical mastoid cavity infection and chronic or relapsing otitis externa. A total of six publications were included in the final evaluation. Enoxacin was effective in 35%; ciprofloxacin (five publications) was used in 82 patients with 67% effectivity in otitis externa and otitis media and 61% effectivity in radical mastoid cavity infection. No serious adverse reactions were reported. The promising efficacy of otitis media and otitis extern and safety profile needs further confirmation in double blind prospective clinical studies, that will provide a firm basis for changing the current treatment schedules of COS.

Des biopsies gastroduodénales ont été prélevées, tant en sites pathologiques que sur muqueuse normale, chez 128 patients soumis à un examen endoscopique.

In a pilot study 29 cases of suppurative otitis were treated with ciprofloxacin. Acceptable clinical results were obtained in case of chronic otitis media without cholesteatoma and in case of otitis externa. Cholesteatoma and mastoid cavity cases gave no satisfactory results. Guidelines are proposed to define the specific indications and place of this drug in suppurative otitis.

Seventeen strains of mycobacteria, recovered from six armadillos experimentally infected with Mycobacterium leprae, were examined in ten different laboratories. This collaborative study included use of conventional bacteriological tests, lipid analyses, determination of mycobactins and peptidoglycans, characterization by Py-MS, and immunological, metabolic, pathological and DNA studies. These armadillo-derived mycobacteria (ADM) formed five homogeneous groups (numbered ADM 1 to 5) on the basis of phenetic analyses. However, DNA studies revealed only four homogeneous groups since group ADM 1 and one of the two strains in group ADM 3 showed a high level of DNA relatedness. The phenetic and DNA studies confirmed that the ADM strains differed from all other known mycobacteria. Cultural, biochemical, metabolic and pathogenic properties as well as DNA-DNA hybridizations clearly differentiated these ADM from M. leprae.

The second cooperative numerical taxonomic analysis of Runyon group IV rapidly growing mycobacteria was facilitated by combining the results of 12 individual participants, each performing one or more of the selected tests by his or her own choice of method. The 177 characters were coded for analysis and the matching matrix was generated. Eight of nine clusters which were defined in this investigation were of previously named species. The recognized clusters were as follows: Mycobacterium flavescens, M. vaccae, M. chitae, M. thermoresistibile, and M. smegmatis. One large cluster, designated “M. parafortuitum complex” was noted which contained strains representative of taxa earlier identified as M. parafortuitum, M. diernhoferi, M. aurum, M. neoaurum, and a cluster of strains of unnamed species. It is suggested that segregation of species or subspecies within this complex not be attempted at this time.

This is a report of the second cooperative study, conducted by the International Working Group on Mycobacterial Taxonomy, to standardize and establish reproducibility of tests for classifying and identifying mycobacteria. Five tests that met specified rigorous criteria have already been described (Wayne et al., 1974). In the present report, six additional tests are described which meet these criteria. They are tests for production of niacin and of catalase and for resistance to 10 μg of isonicotinic acid hydrazide (isoniazid) per ml, 1 μg of thiophene 2-carboxylic acid hydrazide per ml, 250 μg of hydroxylamine hydrochloride per ml, and 500 μg of p-nitrobenzoate per ml.

Summary: A co-operative taxonomic study has been performed on cultures belonging to the ‘rhodochrous complex’. Phenetic data on 98 cultures (78 rhodochrous cultures, 12 marker cultures with the genus designation Mycobacterium and 8 with the designation Nocardia) studied in four laboratories were collected and analysed by numerical taxonomic methods. The precipitinogenic properties and the presence of different types of mycolic acids were analysed independently to establish correlation with the numerical classification. The chemotaxonomic and serological data correlated well with the numerical analyses. The rhodochrous taxon can be distinguished from the genera Mycobacterium and Nocardia, and from the proposed genus Actinomadura; furthermore, it can be divided into at least three homogeneous subgroups. Further studies however, are needed before the question of the generic location of the rhodochrous taxon can be settled and before the taxonomic status of the rhodochrous subclusters can be resolved.

The International Working Group on Mycobacterial Taxonomy has undertaken a series of cooperative studies to standardize and establish reproducibility of tests that are useful for classifying and identifying mycobacteria. To date 25 techniques have been examined, and 5 of these met our rigorous criteria for reproducibility and differential power. The properties determined by these tests are urease activity, pigment production, tolerance to 5% NaCl, hydrolysis of Tween 80, and galactosidase activity.

SUMMARY A co-operative study of 28 cultures of Mycobacterium fortuitum by three laboratories, each using different methods, is described. Two agglutination types, four sensitin types, five immunodiffusion types, three lipids types and three biotypes were recognised amongst the strains. In most cases there was good agreement between different means of typing, and the strains could be divided into three groups. Twelve strains, including the type strains of Myco. ranae and Myco. fortuitum, formed a completely homogeneous group as judged by all methods used. A second and slightly less homogeneous group of seven strains included the type cultures of Myco. giae and Myco. peregrinum, and a third group, the least homogeneous, contained nine strains of unnamed organisms. It is suggested that the first two groups might be considered subspecies, Myco. fortuitum fortuitum and Myco. fortuitum giae (peregrinum) respectively.

SUMMARY: A co-operative taxonomic study has been performed on slowly growing non-pigmented mycobacteria (Runyon's group III). Phenetic data on 89 strains, studied in 18 laboratories, were collected and analysed by a numerical taxonomic method. A variety of immunological properties, lipid analyses and measures of pathogenicity were analysed independently to establish correlation with numerical classification. Mycobacterium gastri, M. nonchromogenicum, M. terrae, M. avium and M. xenopi were recognized by almost all participants as distinct species. Mycobacterium novum was considered to be synonymous with M. terrae. A clearcut distinction could not be made between M. avium and M. intracellulare; the majority of participants in the study recommend that M. intracellulare be reduced to a synonym of M. avium. A minority of authors cannot agree with this proposal.

SUMMARY: A co-operative numerical taxonomic analysis of rapidly growing mycobacteria of Runyon's group IV is reported. There was no limitation on the number, nature, or method of performance of the test characters contributed by each of the 12 participants. Initially 415 test characters were coded for analysis; deletion of irrelevant and repetitious data resulted in a final 195 characters used to generate the matching matrix. All nine major clusters defined in the study were of named species; however, three of these were noted to contain two or more species and the reduction of some of these to synonymy with prior epithets is discussed. Recognized clusters were: Mycobacterium smegmatis, M. phlei, M. vaccae (including M. para-fortuitum), M. diernhoferi, M. flavescens, the rhodochrous taxon, M. thamnopheos, M. fortuitum, M. chelonei. Results of the pooled information are compared with those of the individual participants. Immunological results generally correlated well with numerical analyses.

SUMMARY: A co-operative numerical taxonomic analysis of slowly growing scotochromogenic mycobacteria of Runyon′s group II with no restrictions on numbers of characters contributed by individual participants, or on techniques employed to derive these characters, is reported. Of four scotochromogen clusters defined in this study, three were of named species (Mycobacterium flavescens, M. gordonae and M. scrofulaceum) and one, a small cluster of four strains, may represent a new species. Results from pooled data are compared to results obtained with data from individual participating laboratories. Immunologic data were treated separately and showed good correlation with the numerical analyses.

During the past several years, the Mycology Unit of the Communicable Disease Center has had an opportunity to study a number of dermatophytes isolated from various parts of Africa. Among these, Trichophyton yaoundei, a new species described by Cochet, Doby-Dubois, Deblock and Vaiva in 1957 (1), merits special consideration, as it is little known and its specific identity has not been confirmed by other workers T. yaoundei, originally isolated from patients in the former French colony of Cameroon, was first described as T. camerounense by Cochet and Doby-Dubois in 1957 (2). Later in the same year, these authors along with Deblock and Vaiva renamed the organism T. yaoundei since they had found that the earlier combination had been used to describe a different organism. The specific name yaoundei was derived from the province of Yaounde where the organism was first isolated. The authors stated that this species may have been observed previously by Catanei in 1939 (3) when he described a “prune-colored” dermatophyte among a group of isolates obtained from various African countries. In 1955 J. N. Doby and M. Doby-Dubois conducted a routine survey for tinea capitis among 3,000 school children, both Caucasian and Negro, in the province of Yaounde. Of 1,400 children with scajp lesions, 866 were diagnosed as having ringworm infections on the basis of cultural studies. All the infections were found among the children of the Bantu Bamiléké and Haoussa tribes. Of these cases 80.8 per cent were due to various trichophyton species and 19.2 per cent were due to microsporum species. Among the trichophytons, 241 (49.6%) were recognized as a new trichophyton species, later described as T. yaoundei. The other trichophyton species isolated were T. sudanense Joyeux, 1912, 149 strains (30.7%), and T. jerrugineum\ Ota, 1922, 64 strains (13.1%). In 9 instances, T. yaoundei was found in combined infections with other dermatophyte species. In unmixed infections the hairs were observed to be invaded in an endothrix pattern with large spores. MATERIALS AND METHODS Among the African dermatophyte isolates studied at the Mycology Unit, 7 cultures have been identified as T. yaoundei. These cultures and their sources are listed in Table I. In one instance cultures were isolated at the Mycology Unit directly from hairs sent from Africa (Stanleyville, Belgian Congo). Of the remaining cultures, six were isolated in the Belgian Congo from patients observed at the Laboratoires Médical Provincial of both Stanleyville and Elisabethville.** When infected hairs were available, some were studied microscopically and others were cultured. All strains were grown on Sabouraud dextrose agar, casein agar, wort agarjf and rice grains. Nutritional tests as used for the identification of dermatophytes (5) were carried out. The isolates were grown on hair in vitro (6) and on If T. ferrugineum is considered to be a member of the genus Microsporum by myself (Lucille K. Georg) and a number of other mycologists. Isolated by P. Dupagne, 1958. ** Isolated by S. R. Pattyn, 1959. ft Casein Agar: Casamino acid (vitamin free) 2.5 gm. Glucose 40.0 gm. MgS04 0.1 gm. KH2PO4 1.8 gm. Agar 20.0 gm. Water (distilled) 1000.0 ml. (Adjust pH to 6.8) 19 20 THE JOURNAL OF INVESTIGATIVE DERMATOLOGY W < 4 * •**' -\ -TV J* : jy** y y % IS!pliiIll MB Plate I. a.Trichophyton yaoundei, Strain B253. Colony on Sabouraud dextrose agar at 8 weeks, b.T. yaoundei, Strain B253. Close-up of colony, c.T. yaoundei, Strain B124. Close-up of colony, d.T. yaoundei. Mycelium from glabrous colony X 256. e.T. yaoundei. Glabrous colonies with brown pigmentation. One colony showing development of area of white fluffy growth, f.T. yaoundei, Strain B309. Microconidia from growth on casein agar, X 512. TRICHOPHYTON YAOUNDEI 21 ' PLATE H. aHuml hair infection byVs-, Shows endothrix pattern of invasion. X b. Edge of hair infected by 2\yaoundei, X 475. moistened soil baited with hair (7). In addition, animal inoculation studies were carried out. RESULTS I.Study of Clinical Material In one instance infected hairs were available for study. They were in the form of short, dark fragments usually bent or curved. In KOri mounts it was observed that they were invaded in an endothrix pattern. The short hair fragments were broken off bluntly at each end. They were filled with masses of large spores (2 1-7 3 m) with an avera e slze of 4'7 M ( , I a and b). After remaining in the alkaline solution for an hour or longer, the hairs swelled and their walls ruptured. The spores which emerged were observed to be in chains. Cultures made from remaining hairs from this specimen yielded T. yaoundei. II.Morphological Studies of Fungus Isolates a. Gross colony appearances All isolates developed very slowly at room temperature on Sabouraud dextrose agar or on Cycloheximide Medium (8). Cycloheximide did not inhibit their growth. The colonies first appeared after 7 to 10 days of incubation as small, dull white, or slightly yellowish growths with a moist or glabrous to wax-like surface. Their surface was at first either flat or raised m the form of small knobs. However, most colonies became definitely heaped and irregularly folded, particularly at the center of the colony. In * Average spore size derived from measurement of 100 arthrospores. some cases the central area became depressed or the surface of the colony had an irregular lumpy appearance with many knobs and depressions. Only 3 of the strains remained flat, however, these developed shallow radial grooves or a finely wrinkled or verrucose surface. Colonies obtained by inoculating infected hairs only attained a diameter of about 10 mm. after 4 to 6 weeks of incubation. Maximum colony size after 10 weeks incubation was 30 mm. diameter. Subcultures developed more rapidly than the original isolates. In the early stages of development-up to 6 to 4 weeks growth-the colonies were very similar to those of T. verrucosum, and could easily be mistaken for that organism. They resembled also the glabrous, non-pigmented form of 1. violaceum (var. glabrum). Upon prolonged incubation however, the colonies of T yaound™ developed a characteristic tan to dark brown pigmentation. In most of the isolates, pigmentation developed only after 4 weeks incubation at which time the colonies assumed a tan coloration. Some strains remained yellowish tan but others gradually darkened and eventually became deep chocolate brown. Some strains required 8 to 10 weeks incubation before the deepest brown was attained. The majority of the strains produced a tan pigment on the reverse side of the colony also. This pigment was soluble and diffused throughout the medium. In the original description of the Cameroon strains by Cochet et al (1), all cultures became tan or dark chocolate brown, and the medium was always filled with brown pigment. Plate II illustrates the gross morphology of recently isolated strains. 22 THE JOURNAL OF INVESTIGATIVE DERMATOLOGY TABLE I List of isolates of T. yaoundei studied Stock No. Senders' No. B250 387 B251 537 B252 887 B253 84 B289 124 X247 30 B308 617 B309 633 B310 646 B311 708 Type of Material Culture Culture Culture Hair Hair Hair Culture Culture Culture Culture Tinea capitis, Cameroon Tinea capitis, Cameroon Tinea capitis, Cameroon Tinea capitis, Stanleyville, Belgian Congo Tinea capitis, Stanleyville, Belgian Congo Tinea capitis, Stanleyville, Belgian Congo Tinea capitis, Elisabethville,* Belgian Congo Tinea capitis, Elisabethville, Belgian Congo Tinea capitis, Elisabethville, Belgian Congo Tinea capitis, Elisabethville, Belgian Congo Sender of Culture or Hairs G. Cochet G. Cochet G. Cochet P. Dupagne P. Dupagne P. Dupagne S. R. Pattyn S. R. Pattyn S. R. Pattyn S. R. Pattyn * The isolation of T. yaoundei from Elisabethville and its environs was reported by Pattyn and Sassen in 1960 (4). Most of the strains studied at the CDC had a marked tendency to become polymorphic after several transfers. Not only did individual colonies of an isolate vary in rate of growth, form, and pigmentation, but single colonies developed many different types of growth. Frequently the raised colony was surrounded by flat areas of tough, colorless, glabrous growth; or certain areas of the colony became covered with short white velvety aerial mycelium. Old cultures often were covered completely with white, velvety, aerial mycelium. Cultures which remained glabrous maintained some tan pigmentation. However both the pigmentation of the colonies as well as the diffusible brown pigment became partially or entirely lost after several subcultures. Of the cultures obtained from Dr. Cochet, all grew slowly and formed slightly heaped and folded colonies. Two of the strains had glabrous areas with a light tan pigmentation and some areas covered with a white velvety aerial mycelium. One of the strains was completely overgrown with white, velvety mycelium. None of these strains became dark brown and no brown pigment developed in the medium surrounding these colonies.* Growth of our isolates on enriched media such as casein agar, and brain heart infusion agar, appreciably changed the appearance of the * According to a communication from Dr. Cochet, these strains originally exhibited the characteristic chocolate brown pigment of this species. However, after several transplants, this pigmentation has completely disappeared. colonies. Growth was more rapid, and development of the white, velvety aerial mycelium occurred earlier and covered most of the colony. The characteristic chocolate brown, glabrous colonies were not obtained on these media. Growth on wort agar was similar to that on Sabouraud dextrose agar. All strains grew at approximately the same rate whether incubated at 25°C or 37°C. The isolates grew very slowly on rice grains, producing only a small amount of white aerial mycelium after several months incubation. No growth was obtained on the soil-hair plates. This medium, designed to induce sexual reproduction, consisted of autoclaved soil that had been moistened with water and baited with sterilized bits of horse hair (7). b. Microscopic appearance The colonies were tough and leathery in texture. Teased preparations in lacto-phenol cotton blue revealed a branched mycelium of irregular diameter which frequently broke up into short segments. The drier parts of the colonies were friable and composed of masses of chlamydo-spores. The more or less submerged thin, glabrous growth at the edges of the colonies was composed of very delicate, thin mycelium which was more regular in form. Attempts to make mounts of the aerial growth were usually unsuccessful unless slide culture preparations were made. Slide culture preparations made with Sabouraud dextrose agar revealed a thick irregular non-sporulating mycelium. However, in casein agar slide cultures, a fine branched my- TRICHOPHYTON YAOUNDEI 23 celium of uniform diameter that bore numerous delicate microconidia developed. These were small, pyriform, hyaline, one-celled, smooth walled spores borne laterally on the aerial mycelium and attached directly by their slightly truncate bases. Some terminal clusters of spores also developed. The conidia were approximately 1.5 to 2.5 jLt long by 0.75 x 1.0 /x wide. In general they were similar to the microconidia of other trichophyton species. Hitherto, microconidia had not been described for this species. Macro-conidia were not found in any preparation. Rice grains, wort agar and brain heart infusion agar did not stimulate sporulation. Plate II (d and f) illustrates the microscopic morphology of T. yaoundei. III.Physiological Studies Nutritional studies were undertaken according to the methods described by Georg and Camp (5). All strains were found to have the ability to develop equally well on vitamin-free media as on vitamin containing media. Growth was consistently better on casein, vitamin-free agar than on ammonium nitrate, vitamin-free agar. In vitro hair tests as described by Ajello and Georg (6), were tried with all strains. The organisms grew well in the dilute yeast extract solution; however, no growth occurred on the hairs, and accordingly perforations were not observed after 4 weeks incubation. IV.Animal Inoculations Attempts to produce experimental ringworm infections in guinea-pigs were unsuccessful. DISCUSSION Trichophyton yaoundei appears to be a distinct dermatophyte species. It is characterized by the development on Sabouraud dextrose agar of irregularly heaped glabrous colonies which are at first cream to tan in color, but after several weeks incubation, become a dark chocolate brown. During growth of the colony, the medium becomes diffused with a soluble brown pigment. The organism grows well on a vitamin-free medium such as casein agar. It develops slowly on vitamin-free NH4N03 agar. Its growth is not stimulated by the addition of vitamins or incubation at 37°C. Microconidia characteristic of the genus trichophyton are developed by some strains on casein agar. This species was apparently first encountered by Cochet and Doby-Dubois in the French Cameroon in 1957. It is doubtful that the “prune colored” dermatophyte isolated from the Cameroon by Catanei in 1939 represented T. yaoundei (as suggested by Cochet and Doby-Dubois) since he made no mention of a deep browrn or “chocolate colored” pigment. T. yaoundei is distinguished from the morphologically similar T. verrucosum by the following. 1.T. verrucosum grows much more slowly on Sabouraud dextrose agar. 2.T. verrucosum grows better at 37 °C than at room temperature. 3.T. verrucosum has complete requirements for one or more vitamins (thiamine, inositol, pyridoxine), and will not grow on vitamin-free media (9). 4.T. verrucosum does not develop dark brown pigmentation. 5.T. verrucosum produces a large-spored ecto-thrix type of hair invasion. T. yaoundei is different'ated from T. violaceum by the following. 1.T. violaceum usually develops some lavender or purple pigmentation. 2.T. violaceum, as well as its variety glabrum, never develops colonies with dark brown pigmentation. 3.T. violaceum will develop very slowly on vitamin free casein agar, but is greatly stimulated by the addition of thiamine to this medium (10). Clinically T. yaoundei infections appear to be similar to those produced by other endothrix species such as T. violaceum, T. tonsurans, and T. sudanense. T. yaoundei apparently has a limited geographic distribution. At present it has been reported only from the Cameroon Republic, and from the Republic of the Congo. STRAINS AVAILABLE Three strains of T. yaoundei from this series have been deposited with the American Type Culture Collection. The culture collection numbers are listed below. ATCC 13946 Trichophyton yaoundei. Isolated at the Mycology Unit, Communicable Disease Center, Atlanta, Georgia, from endothrix 24 THE JOURNAL OF INVESTIGATIVE DERMATOLOGY hairs, Stanleyville, Belgian Congo (Dec. 3, 1958) CDC #B289. ATCC 13947 Trichophyton yaoundei. Isolated by Dr. S. R. Pattyn from tinea capitis, Elisabethville, Belgian Congo (Aug. 15, 1959) CDC #B309. ATCC 13948 Trichophyton yaoundei. Same source as 13947 (Aug. 15, 1959) CDC #B311. SUMMARY 1. Seven strains of T. yaoundei from several areas in Africa were studied and described. 2.T. yaoundei appears to be a distinct dermatophyte species. It has been isolated from at least 250 cases of endothrix tinea capitis in African natives. Cultures from infected hairs develop slowly on Sabouraud dextrose agar. The colony is irregularly heaped and folded and is largely glabrous with a wax-like surface. It is at first cream to tan, but after several weeks becomes a dark chocolate brown. The medium becomes filled with a diffusible brown pigment. 3. Microconidia were found for the first time in this species by growing the isolates on casein agar. 4. Criteria useful in the identification of this species, and for its differentiation from morphologically similar species were defined.