Effects of surface active agents on the susceptibility of Swiss mice to Candida albicans

Summary The LD50's ofCandida albicans combined with Plurafac B26, Mulsor 224, Pluronic L62, and Polyethylene Glycol 400 Mono Laurate injected intraperitoneally into female white Swiss mice were calculated by the method ofReed &Muench. Each of the previously mentioned surface-active agent —Candida albicans combinations had a lower LD50 than theCandida albicans control.Plurafac B26 was found to intially decrease the number of leukocytes in the peritoneal cavity and thus enhance the invasiveness ofCandida albicans.     

Quelques champignons pathogènes,isolés du sol roumain,par la méthode de l'inoculation intrapéritonéale à la souris

Resumé En injectant, selon le procédé d'Ajello et al., dans la cavité péritonéale de la souris blanche, une suspension de terre prélevée de divers endroits et en ensemençant après un délai de 6 à 8 semaines des fragments du foie, de la rate, des poumons et des reins de l'animal inoculé, les auteurs ont décelé dans le sol roumain la présence des champignons suivants:Geotrichum candidum, Trichophyton mentagrophytes, Keratinomyces ajelloi (une souche pathogène) etCoccidioides immitis. Selon les auteurs, cette méthode d'isolement est capable de compléter la méthode deVanbreuseghem pour l'isolement des dermatophytes du sol. La présence duCoccidioides immitis dans le sol roumain mérite une mention spéciale, la coccidioidomicose étant, jusqu'en 1961, inconnue en Roumanie.

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Summary By injecting the supernatant from aqueous suspensions of different rumanian soil specimens intraperitoneally into white mice and subsequent culture of their livers and spleens, according to the procedure ofAjello et al., the authors were able to recover the following fungi:Geotrichum candidum, Trichophyton mentagrophytes, Keratinomyces ajelloi (a pathogenic strain) andCoccidioides immitis (two strains). In the author's opinion, this indirect isolation procedure may complete the method developed byVanbreuseghem for isolating dermatophytes from soil. The presence ofCoccidioides immitis in the rumanian soils needs special mention owing to the fact that coccidioidomycosis was unknown in Rumania until 1961.

     

The effects of culture media on Saprolegnia antigens

In a previous paper (Hodkinson &Hunter, 1970a) we reported that salmon, suffering from a disease called ulcerative dermal necrosis, appeared to have precipitating antibodies to the mycelium of a fungus associated with their diseased skin lesions. This was confirmed when antigens were prepared from pure cultures of the fungus, a species ofSaprolegnia (Hodkinson &Hunter, 1970b). In this paper we describe the effects of culture media on the antigenicity ofSaprolegnia mycelium, as detected by double gel diffusion with salmon sera.     

Phage typing ofSalmonella Typhi in the Netherlands

Summary A survey has been given of the results of phage typing of strains ofS. typhi found in Holland. It has been shown, that type A includes a different group of strains in systems drawn up with different Vi phages. An auxiliary system — to be used besides the system ofCraigie andYen — and a few new types, have been described.     

Taxonomic status of the species of the green algal genusNeochloris

Komárek has recently reviewed the various species assigned to the green algal genusNeochloris Starr (Chlorococcales, Chlorococcaceae) and removed those with uninucleate vegetative cells to a new genus,Ettlia. Watanabe & Floyd, unaware ofKomárek's work, also reviewed the species ofNeochloris and distributed them among three genera—Neochloris, Chlorococcopsis gen. nov., andParietochloris gen. nov.—on the basis of details of the covering of the zoospore and the arrangement of the basal bodies of the flagellar apparatus. This paper reconciles these two treatments and makes additional recommendations at the ranks of genus, family, order, and class.     

Taxonomy of Haplosporangium parvum

Summary A strain ofHaplosporangium parvum has been isolated from Italian soil. It has been comparatively studied with six North American and a Korean strain. Two American and the Italian strains are thermophilic (opt. about 35° C) while the other four grow better at temperatures around 20° C.The assimilation of nitrogen sources has been comparatively studied. In addition to peptoneH. parvum metabolizes urea and most aminoacids, whileH. bisporale assimilates peptone, leucine, alanine, glutamic acid and proline only. On the basis of N assimilation and thermophily, a key of the studied strain is established.A comparison ofH. bisporale withH. parvum, confirmed that the former species is a Phycomycete, while the latter is a Moniliaceous, aleuriosporic fungus. All the strains are feebly pathogenic, but the virulence of the Italian soil isolate is equal to that of the most virulent strains isolated from animal lungs (Alberta, Canada).The taxonomy ofH. parvum has been studied in relation to the more or less allied generaAleurisma, Glenosporella, Geomyces andStaphylotrichum.It is concluded thatH. parvum may be better classified as a new genus, and the monotypic genusEmmonsia withE. parva (Emm. etAshb.) n. comb. is proposed, although the entire group of aleuriosporaceous fungi needs revision.     

Inhibition of in vitro germination and spherulation ofCoccidioides immitis byAllium sativum

An aqueous extract of a dehydrated garlic preparation with uniform consistency inhibited all eight clinical isolates of the dimorphic fungus,Coccidioides immitis. The inhibitory and lethic concentrations were in the range of 3.12–6.25 mg/ml for both the saprophytic (mold) and parasitic (spherule) forms ofC. immitis. At 6.25-mg/ml concentration, the organism lost its viability within 6 h. The conversion of arthroconidia into spherules in a chemically defined liquid medium was prevented by garlic extract diluted to 1:320 (3.12 mg/ml). The data indicate that components of garlic readily inhibited the in vitro germination and spherulation of this medically important dimorphic fungus.     

The occurrence of trichophyton indicum — Randhawa & Sandhu 1963 in rumanian soil. Some data concerning its immunobiological properties. (Preliminary report)

Summary Studies are carried out on the presence ofTrichophyton indicum —Randhawa &Sandhu 1963 in Rumanian soil and on some of its immunobiological properties. Recently the authors were able to demonstrate the presence of this fungus in Rumania, in soil samples from 4 different locations. In spite of the unsuccessful attempts to inoculate it with positive result in men and in test animals, the immunobiological properties of this fungus proved characteristic for a dermatophyte.

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Résumé Les auteurs ont réussi à mettre en évidence dans le sol roumain la présence duTrichophyton indicum —Randhawa &Sandhu 1963. Ce champignon a été isolé à partir d'échantillons de terre provenant de 4 régions différentes. Malgré la faillite des essais d'inoculation de ses cultures à l'homme et à l'animal, les propriétés immunobiologiques duT. indicum sont celles d'un dermatophyte vrai.

     

The effect of fungizone on experimental adiaspiromycosis of laboratory mice

Summary In laboratory mice weighing about 25 g, with experimental infection of adiaspiromycosis, total inhibition of development of the parasitic phase of the agent of this mycosis is induced with Fungizone — Squibb in daily intraperitoneal administration of 0.03 mg (dissolved in 0.1 ml of isotonic saline). In autopsies carried out 10th, 20th and 30th day after an intraperitoneal inoculation of 1 ml suspension of the mycelial phase of two Czechoslovak isolates ofEmmonsia crescens Emmons &Jellison 1960, no elements of the parasitic phase — adiaspores were found in the organs of the treated mice. In control groups of the untreated mice, experimental infection developed which was demonstrated by macro- and microscopical controls of organs and by a successful recovery of the agent.     

Parthenogenetic production of fertile Cleistothecia in Nannizzia incurvata stockd. Under the influence of Chrysosporium species

Summary The parthenogenetic elicitation of mature, fertile asci and ascospores inNannizzia incurvata,Stockd. (Sabouraudia,1: 46, 1961) and inMicrosporon gypseum,Guiart &Grigoraki, 1928 by mating them withChrysosporium species is described. The same effect was obtained when culture filtrate ofChrysosporium species alone was used instead of the living fungus. In ascospore-derivatives (ascoagarblock) profound morphological changes were observed as to the shape and size of the macro- and microconidia. The peridial ornaments showed alone or in mixture those ofNannizzia fulva, Nannizzia gypsea along with the ornaments ofNannizzia incurvata. The derivatives of parthenogenetic ascospores were different from the mother-species macro- and microscopically and remained such through many generations as stable mutants.

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Zusammenfassung Die parthenogenetische Auslösung von fruchtbaren Asci und Askosporen inNannizzia incurvata Stockd. (Sabouraudia,1: 46, 1961) und inMicrosporon gypseum,Guiart &Grigoraki, 1928 durch Kreuzung mit Arten vonChrysosporium wird beschrieben. Dasselbe Ergebnis wurde erzielt, wenn — anstatt des lebenden Pilzes — das Kulturfiltrat benützt wurde. In den Abkömmlingen der parthenogenetisch erhaltenen Askosporen (Asko-Agarblock) wurden tief gehende, morphologische Veränderungen betreffs Gestalt und Grösze der Makro- und Mikrokonidien beobachtet. Die peridialen Ornamente zeigten eine Mischung derjenigen vonNannizzia fulva, undNannizzia gypsea mit denen vonNannizzia incurvata. Die Abkömmlinge der parthenogenetisch erhaltenen Askosporen waren makro- und mikroskopisch verschieden von den originalen Stämmen und sie verblieben so durch viele Generationen als stabile Mutanten.

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Résumé La production parthénogénétique des asques et des ascospores fertiles enNannizzia incurvata Stockd. (Sabouraudia,1: 46, 1961) et enMicrosporon gypseum,Guiart &Grigoraki, 1928 par croisement avec des espèces deChrysosporium est décrite. On a obtenu le même résultat avec des filtrations des cultures liquides deChrysosporium, sans la présence du champignon vivant. Les dérivés des ascospores parthénogénétiques présentent un changement morphologique profond quant'à la forme et à la grandeur des macro- et microconidies. Des ornements peridiales présentent une combinaison de ceux deNannizzia fulva, Nannizia gypsea et deNannizzia incurvata. Les dérivés des ascospores parthénogénétiques sont différents en leur forme macro- et microscopique de l'espècemère, et sont restés sans change par de nombreuses générations comme une mutation stabile.

     

Considérations à proposdes caractères mycologiques et pathogéniques du Keratinomyces ajelloi — Vanbreuseghem 1952, saprophyte du sol

Summary The authors report, for the first time in Rumania, the presence in the soil ofKeratinomyces ajelloi —Vanbreuseghem 1952, isolated by them on 3 different places at Bucharest. One of the isolated strains showed a little more abundant microscopical morphology than the strain first isolated byVanbreuseghem, for it had also coils and chlamydospores.Although this new dermatophyte has been considered, up to now, nonpathogenic for man and animal, the authors were able to obtain positive results in white mice, guinea pig and in man, inoculating repeatedly one of the isolated strains. The experimental lesion it produced, of herpes circinatus type, disappeared in a few days.As the authors conclude, it is not necessary to create a new genus for this dermatophyte. Considering its microscopical aspect, its pathogenicity and its soil origin, they propose to classifyKeratinomyces ajelloi in the genusEpidermophyton —Ota etLangeron 1923 —, giving it the name ofEpidermophyton terrigenum.

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Résumé Les auteurs ont signalé, pour la première fois en Roumanie (à Bucarest), la présence dans les couches superficielles du sol duKeratinomyces ajelloi —Vanbreuseghem 1952. La morphologie microscopique de l'une des deux souches, étudiées de ce point de vue, était un peu plus riche que celle de la souche isolée, pour la première fois parVanbreuseghem car, outre les macroconidies, on y trouvait des vrilles serrées et des chlamydospores. D'autre part, chez les deux souches, les extrémités libres des macroconidies n'étaient pas constament amincies et, par places, plutôt arrondies. L'une des souches s'est montrée pathogène pour l'homme, le cobaye et la souris blanche, chez lesquels son inoculation experimentale a provoqué une mycose superficielle authentique, bien que passagère. Ce caractère n'a pas été signalé jusqu'à présent. Vue sa morphologie microscopique, sa pathogénité et son origine tellurique, les auteurs proposent, pour cette espèce, le nom d'Epidermophyton terrigenum.

     

Morphological and cytological investigations ofUlothrix zonata and taxonomy of the related species

The present communication deals with the observations made on the morphology, reproduction and cytology ofUlothrix zonata (Weber & Mohr)Kütz. in culture. The alga displays a remarkable phenotypic plasticity in nature as well as in culture. The present study provides additional evidence of karyology in support ofLokhorst andVroman's treatment ofU. zonata which merges a number of earlier described species in it. The cytological details and chromosome number (n = 10) determined for the Indian isolates ofU. zonata agree with those ofSarma for the British material.NBRI Research Publication No. 63 (N.S.)     

Growth factor requirements of two strains ofSphaerotilus discophorus

The relatively complex growth-factor requirements of two strains ofSphaerotilus discophorus, strains 31 and 32, have been elucidated. In addition to thiamin and biotin which are required by other strains ofS. discophorus, the two strains must be supplied also with adenine or guanine and with cyanocobalamin for growth in a glucose — Casamino Acids — mineral salts medium. The cyanocobalamin can be replaced by methionine but only if relatively large amounts of this amino acid, 100 µg or more per ml, are added to the medium. The methionine requirement of the two strains is approximately 3 times greater than that of otherS. discophorus strains.     

Blastomyces cerolytica (sp. n.) and its relation to Coccidioides

Summary In 1949 I discovered a yeast like fungus which was termedBlastomyces cerolytica. During the essay for exact determination of this fungus, it was found that it resemblesCoccidioides. Its yeast-like form is zymatic, malto-saccharasic. As organs of fructification aleurospores and chlamydospores were found, as well as asci containing hundreds af ascospores. It shows isogame heterothallism. It belongs to Taphrinales. A comparison between characteristics ofCoccidioides immitis andBlastomyces cerolytica is given. The diagnosis of this fungus concludes the investigation.

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Resumé En 1949 nous avons découvert un champignon levuriforme, lequel nous avons nomméBlastomyces cerolytique. Il nous a paru d'être très semblable au genreCoccidioides. Son état levuriforme se comporte comme une levure zymatique et malto-saccharasique. Les formes asexuelles sont des aleurospores et des chlamydospores et les formes sexuelles des asques géants remplis de centaines d'ascospores. Il paraît être isogame hétérothallique. Il appartient à des Taphrinales. Une comparaison était mise en vue des caractéristiques deCoccidioides immitis et deBlastomyces cerolytique. Nous donnons aussi une diagnose pour ce champignon.

     

Inhibition of the mold phase of Coccidioides immitis by beta-ionine and dimethoxane

The mold phase ofCoccidioides immitis was exposed to varying concentrations of beta-ionone and dimethoxane in Sabouraud'sl Glucose Agar and the inhibitory concentration of each compound was determined. Beta-ionone demonstrated an inhibitory effect at 0.2 % or less, but completely inhibited the growth ofC. immitis above 0.2 %. Its mouse LD₅₀ was 4.4 × 10⁴ mg. Because of its great toxicity for mice, it could not be considered a possible candidate for chemotherapy. However, results suggested its use as a selective additive for an isolation medium. Dimethoxane demonstrated complete inhibition ofC. immitis at concentrations of 0.55 % and higher and some fungistatic effects at lower concentrations. Because the mouse IP toxicity dose was found to be rather low (0.65 mg/kg), it may have potential as a chemotherapeutic agent for coccidioidomycosis.     

A rapid pigmentation test for identification ofCryptococcus neoformans

A rapid pigmentation test for identification ofCryptococcus neoformans is described. The method is based on the formation of a characteristic, mousegrey to violaceous-black pigment when shake cultures ofC. neoformans in a phosphate-buffered,l-DOPA — ferric citrate medium are incubated at 37 C for one hour.This work was carried out in part with financial support from the Council of Scientific and Industrial Research, New Delhi.     

Quantification of the histochemical reaction for alkaline phosphatase activity using the indoxyl-tetranitro BT method

Summary The indoxyl—tetranitro BT method for the demonstration of alkaline phosphatase activity has been optimized and its validity for quantitative histochemistry tested. The study has been performed with model films of polyacrylamide gel incorporating homogenate of rat liver and with cryostat sections from the same livers. Addition of polyvinyl alcohol to the incubation medium greatly improved the localization of the final reaction product in cryostat sections. In polyacrylamide films, the formazan production specifically due to alkaline phosphatase was highest when using a medium containing 100mm Tris-HCl buffer, pH 9.0, 0.2–1.0mm substrate, 0.32mm 1-methoxyphenazine methosulphate, 10mm MgCl₂, 5mm sodium azide and 1mm tetranitro BT. For the incubation of cryostat sections in the presence of polyvinyl alcohol, the same medium could be used but the optimum concentrations of substrate and tetranitro BT appeared to be 1–2mm and 5mm respectively. The test minus control reaction was specific for alkaline phosphatase activity and could be inhibited completely with tetramisole. The test minus control reaction was linear with time up to 30 min with model films and up to 15 min with cryostat sections. The formazan production was also linear with the amount of homogenate incorporated in model films and with section thickness up to 18 µm and therefore, the reaction obeyed the Beer—Lambert law. Variation of the substrate concentration yielded aK _M of 0.05mm for aqueous media and aK _M of 0.55mm for polyvinyl alcohol-containing media. The inhibition with tetramisole appeared to be competitive withK _i = 0.07mm for aqueous media andK _i = 0.7mm for polyvinyl alcohol-containing media. These values indicate that the indoxyl—tetranitro BT method is considerably more sensitive than any metal salt or diazonium salt method developed so far. It is concluded that the optimized method described here is a specific, sensitive and valid quantitative histochemical method for the demonstration of alkaline phosphatase activity.     

Antigenic relationships between Aracoccidioides loboi and other pathogenic fungi determined by immunofluorescence

Summary The fluorescent antibody technique was used to study antigenic relationships betweenParacoccidioides loboi and other pathogenic fungi. The findings suggest thatP. loboi is more closely related antigenically to certainP. brasiliensis strains than to others and that it has antigens in common with the yeast form ofHistoplasma capsulatum, H. duboisii, Blastomyces dermatitidis, Candida albicans and also the mycelial form ofCoccidioides immitis. Serum globulins from 3 cases of keloidal blastomycosis were labelled with fluorescein isothiocyanate. These conjugates showed slight or no reactivity withP. loboi, the yeast forms ofP. brasiliensis, H. capsulatum, H. duboisii andB. dermatitidis, However, they stained brightlyC. albicans, serotypes A and B, the tissue form ofC. immitis and the yeast form ofSporotrichum schenckii. Adsorption of these reagents withC. albicans eliminated all staining except that forS. schenckii. These patients had no history of clinical sporotrichosis.Deceased. Last address: Fundacão Gonçalo Moniz, Salvador, Bahia, Brazil. Requests for reprints should be sent to Dr.William Kaplan.Dr.Miranda is in private practice in Rio de Janeiro, Brazil.     

Effect of prior vaccination on experimental Blastomycosis

Immunogens were found associated with particular fractions prepared from spherules ofCoccidioides immitis (Kong, Levine &Smith, 1963) and from yeast cells ofHistoplasma capsulatum (Salvin &Ribi, 1955). However,Blastomyces dermatitidis, another dimorphic systemic fungal pathogen was shown to elicit a minimal immunogenic response in experimental animals (Kong &Levine, 1967). It was therefore deemed pertinent to study factors which might enhance the resistance of mice to infection withB. dermatitidis.     

Repetitive somatic embryogenesis in peanut cotyledon cultures by continual exposure to 2,4-d

Somatic embryos from immature cotyledons in peanut (Arachis hypogaea) were initiated on media supplemented with 2,4-dichlorophenoxyacetic acid (2,4-d). Over 90% primary embryogenesis and 41–46% repetitive embryogenesis were obtained 12 weeks after initiation by maintaining embryogenic cultures on medium containing 20 mg 1^-1 2,4-d. Maintenance of cultures on medium with 30 or 40 mg I^-1 2,4-d resulted in lower primary and secondary embryogenesis, and proliferation of nonembryogenic callus. Transfer of embryogenic cultures to a secondary medium with 10 or 20 mg I^-1 2,4-d significantly enhanced secondary embryogenesis compared to basal medium without the growth regulator. The use of Murashige & Skoog versus Finer's media had no significant effect on embryogenesis (85–95%), repetitive embryogenesis (11–37%) or mean embryo number. Secondary embryogenesis was also maintained for over one year by repeated subculture of isolated somatic embryos on medium with 20 mg I^-1 2,4-d.Abbreviations B5 Gamborg et al. medium (Gamborg et al. 1968) - 2,4-d 2,4-dichlorophenoxyacetic acid - FN Finer & Nagasawa medium (Finer & Nagasawa 1968) - MS Murashige & Skoog medium (Murashige & Skoog 1962)