Showing papers in "Antonie Van Leeuwenhoek International Journal of General and Molecular Microbiology in 1969"

Leucosporidium gen. n., the heterobasidiomycetous stage of several yeasts of the genus Candida.

Abstract: Investigation of yeasts from Antarctic regions revealed that certain species ofCandida have heterobasidiomycetous life cycles. Two distinct but overlapping groups of species were found: heterothallic and self-sporulating species.Candida scottii is a heterothallic species with the following life cycle: opposite mating types will conjugate and develop a dikaryotic mycelium with clamp connections. Karyogamy occurs in the teliospore which germinates and produces a promycelium. Meiosis takes place in the promycelium, followed by development of haploid sporidia to complete the life cycle. In addition,C. scottii has a self-sporulating phase. From a single cell, in the apparent absence of mating, a uninucleate mycelium is produced that lacks clamp connections. Teliospores, promycelia and sporidia develop that appear similar to those produced from dikaryotic mycelium. The self-sporulating species have life histories similar to the self-sporulating phase ofC. scottii; except that heterothallism has not been observed. Based on these life histories the new genusLeucosporidium is proposed with two heterothallic species (Leu. scottii andLeu. capsuligenum) and five self-sporulating species (Leu. antarcticum, Leu. frigidum, Leu. gelidum, Leu. nivalis andLeu. stokesii. Leu. antarcticum) andLeu. stokesii have not been described under the genusCandida.

Epitheliocystis, a new infectious disease of the bluegill (Lepomis macrochirus).

Abstract: A new infectious, benign and chronic disease, histopathologically distinct from the viral disease lymphocystis, has been found in the bluegill. The symptoms are much enlarged cells in skin and gill epithelium, filled with basophilic granules. By electron microscopy these granules were shown to be organisms with the morphological characteristics ofBedsonia (Miyagawanella). During an attempt to isolate the agent in the bluegill fibroblast cell line BF-2, a cytopathic effect somewhat resembling the lesionsin vivo was found in the first passage. In later passages the agent was lost, but a true virus eliciting a distinctive fusiform cytopathic effect was recovered from these passages. The virus was found to be cytopathogenic also for the rainbow trout cell line RTG-2, but no recognizable disease could be evoked in young bluegills by inoculation of infected cell culture material. While the larger sized, non-viral organism is almost certainly the etiological agent of epitheliocystis, the role of the virus is, therefore, conjectural. Electron microscopy of thin sections has shown the virus to resemble influenza virus in morphology.

The non-oxidative decarboxylation of p-hydroxybenzoic acid, gentisic acid, protocatechuic acid and gallic acid by Klebsiella aerogenes (Aerobacter aerogenes).

Abstract: Klebsiella aerogenes adapted to a chemically-defined mineral salts medium with glucose orp-hydroxybenzoate as sole source of carbon and energy possessed constitutive decarboxylases for gentisate (2,5-dihydroxybenzoate), protocatechuate (3,4-dihydroxybenzoate) and gallate (3,4,5-trihydroxybenzoate) whose pH optima were respectively 5.9, 5.6 and 5.8. A decarboxylase for PHB was induced by PHB in both growing and resting cells; the induction was delayed or inhibited by chloramphenicol and by ultrasonic disruption of the bacteria. Crude ultrasonic preparations of PHB decarboxylase had an optimum pH of 6.0, a Michaelis constant of 4mm and an activation energy of 25,500 cal mole−1 at 28 – 38 C. All four decarboxylations proceeded without O2 and for every mole of phenolic acid decomposed one mole of CO2 and one mole of the corresponding phenol were produced. The effects of ultrasonic disruption of the bacteria suggested that permeability barriers limited the rate of decarboxylation of PHB and 2,5-DHB but not of 3,4-DHB or 3,4,5-THB. During ultrasonic disintegration PHB and 3,4-DHB decarboxylases were retained solely by insoluble centrifugeable particles, whereas 2,5-DHB and 3,4,5-THB decarboxylases were gradually released into solution. The decarboxylation of protocatechuic acid is an essential stage in the assimilation ofp-hydroxybenzoic acid byK. aerogenes, whereas the decarboxylation ofp-hydroxybenzoate itself is an injurious side reaction.

A genetical and biochemical study of chlorate-resistant mutants of Salmonella typhimurium.

Abstract: S.typhimurium can form nitrate reductase A, chlorate reductase C, thiosulfate reductase, tetrathionate reductase and formic dehydrogenase. None of these enzymes are formed in chlorate-resistant mutants. Conjugation experiments showed the presence of a strong linkage between thechl andgal markers of the bacterial chromosome. By deletion mapping the gene ordernic A aro G gal bio chl D uvr B chl A was found. Strains with deletions terminating betweenbio anduvr B or betweenuvr B andchl A have a number of aberrant properties. Though resistant against chlorate they reduce nitrate and form gas. After growth with nitrate they form less nitrate reductase than the wild type which may explain the resistance against chlorate. After growth with thiosulfate they form small amounts of thiosulfate reductase and chlorate reductase C. In crosses between anE.coli Hfrchl + strain and aS.typhimurium chl A strain recombinants were obtained, forming nitrate reductase A and chlorate reductase C. These recombinants do not form gas, which indicates that thechl + gene fromE.coli does not function normally inS.typhimurium.

Mode of action of a bacteriocin produced by Enterobacter cloacae DF13.

Abstract: Enterobacter cloacae DF 13 produces a bacteriocin with killing action onKlebsiella edwardsii var.edwardsii. The degree of sensitivity to the bacteriocin depended on the medium in which the cells were grown and on the bacteriocin concentration used. An excess of bacteriocin (60 K.U./ml) arrested growth in about 60 min. Growth of bacteriocin-treated cultures could be restored by trypsin treatment. In Brain Heart Infusion cultures trypsin rapidly restored bacterial growth even after 60 min of bacteriocin treatment. However, in broth cultures and minimal medium cultures treated with bacteriocin for only 10 min, it took 4 to 5 hr before growth started again. The bacteriocin had little effect on resting cells. Broth-grown cells had about 280 and BHI-grown cells about 340 bacteriocin receptor sites. Bacteriocin DF 13 strongly inhibited protein synthesis after a lag-time of 15 to 60 min depending on the concentration used but had no effect on RNA and DNA synthesis nor on respiration and fermentation. The bacteriocin stimulated RNA synthesis in a leucine-deficient mutant after leucine deprivation.

Systematics of the genus Candida Berkhout: proton magnetic resonance spectra of the mannans and mannose-containing polysaccharides as an aid in classification.

Abstract: The proton magnetic resonance (p.m.r.) spectra of the mannans, galactomannans and mannose-containing heteropolysaccharides of a number of species ofCandida were determined, and compared with the spectra of similar polysaccharides of some species of the sporogenous generaPichia, Hansenula, Endomycopsis, Debaryomyces andSaccharomyces. Similarities among the spectra were used as an aid in classification. TheCandida species studied were placed in four groups: 1) thoseCandida species classified as asporogenous forms of perfect species, 2) those which produce mannose-containing polysaccharides having p.m.r. spectra resembling those of the mannans of known perfect species, 3) members of theCandida parapsilosis group, 4) thoseCandida species which did not ferment glucose and which were not included in one of the other groups.

Growth requirements of Haemophilus vaginalis.

Abstract: A medium, composed of peptone, maltose, dextrose and phosphate buffer, which supports good growth ofHaemophilus vaginalis has been devised. Growth was about equally good with soluble starch substituted for maltose. Optimal growth occurred in the media adjusted to pH 6.8 before sterilization and the media sterilized at 112 C for 12 minutes supported better growth than did corresponding media sterilized at 121 C for 15 minutes. The organisms grew both aerobically and anaerobically. For growth in a semi-defined medium, containing enzymatically hydrolyzed vitamin-free casein, inorganic salts, carbohydrates and six purine and pyrimidine bases, the organisms were found to require the following B-vitamins: thiamine · HCl, riboflavin, niacin (or niacinamide), pteroylglutamic acid, and biotin. They failed to grow in the above medium lacking purine and pyrimidine bases, but the single omission of any one of the six bases (adenine sulfate, guanine·HCl, cytosine, uracil, thymine, and xanthine) except adenine sulfate did not affect growth of either strain tested. One strain failed to grow in the medium lacking adenine sulfate. Since the strains investigated, including the type, do not require X and/or V factors or otherwise definable coenzymelike substances, they do not qualify as members of the genusHaemophilus on the basis of its current circumscription. Thus, reconsideration must be given to the classification of the organisms now known asHaemophilus vaginalis.

Occurrence of inducible and NAD(P)-independent primary alcohol dehydrogenases in an alkane-oxidizingPseudomonas

Abstract: Pseudomonas aeruginosa (strain 473) constitutively contains a soluble NADP-linked dehydrogenase active towards primary alcohols In addition, at least two NAD(P)-independent primary alcohol dehydrogenases can be induced by growing this strain on primary alcohols,α,ω-diols orn-alkanes These inducible enzymes were found to be bound to cellular structures They reduce bovine cytochromec and various dyes, but not oxygen The main difference between the inducible enzymes is their different capacity to oxidize ethanol Noteworthy properties of the enzymes are: 1) the affinities for the straight-chain primary alcohols increase with increasing chain length (tested up to 1-decanol); 2) the affinities decrease when polar atoms or groups are introduced into the alcohol molecule; 3) enzyme preparations as well as intact cells, when provided with a mixture of alcohols, first oxidize the compound with the lowest solubility in water

Enzyme repression in the arginine pathway of Saccharomyces cerevisiae.

Abstract: Enzyme repression in the arginine pathway ofSaccharomyces cerevisiae was demonstrated by comparison of specific enzyme activities in yeast grown with and without arginine in various mineral salts media. Of the enzymes tested only ornithine transcarbamoylase was found to be repressed by exogenous arginine. Acetylornithine-glutamate transacetylase and argininosuccinate lyase were not affected. No relationship between specific enzyme activities and intracellular arginine concentration was observed.

The formation of phenol in the degradation of p-hydroxybenzoic acid by Klebsiella aerogenes (Aerobacter aerogenes).

Abstract: After growth ofK. aerogenes in chemically defined media consisting of mineral salts andp-hydroxybenzoate with or without glucose, phenol was found in the culture fluid at concentrations inhibiting further growth. Bacteria adapted to mineral salts medium containingp-hydroxybenzoate as sole source of carbon and energy produced small but isolable quantities of 3,4-dihydroxybenzoic acid and catechol and oxidized these substances as rapidly asp-hydroxybenzoate. Bacteria adapted to mineral salts medium containing glucose as sole carbon and energy source did not oxidizep-hydroxybenzoate, 3,4-dihydroxybenzoate or catechol. Bacteria adapted to glucose medium or top-hydroxybenzoate medium did not oxidize or utilize phenol as sole carbon and energy source. A metabolic pathway forp-hydroxybenzoate degradation is proposed and the formation of phenol is attributed to a side reaction.

The raffinose fermentation ofSaccharomyces pastorianus andSaccharomyces bayanus

Abstract: Quantitative methods for estimating the extent of raffinose fermentation were applied to cultures ofSaccharomyces pastorianus andSaccharomyces bayanus received from culture collections. Four cultures ofSaccharomyces pastorianus fermented two-thirds only of the raffinose molecule. This was in contrast to recently published reports and confirmed the standard description ofSaccharomyces pastorianus. Two cultures ofSaccharomyces bayanus each consisted of two components, the major component was able to ferment two-thirds raffinose and the minor to ferment raffinose completely. Isolates of the major component could readily mutate to the minor. Three cultures ofSaccharomyces bayanus corresponded to the standard description in their ability to ferment only one-third raffinose. The taxonomic implications of these findings were considered.

Bacteriophages of psychrophilic pseudomonads. I. Host range of phage pools active against fish spoilage and fish-pathogenic pseudomonads

Abstract: A total of 180 bacteriophages active against 37 of 55 psychrophilic bacteria (isolated from fish and other sources) were isolated from sea water, sewage, and fish fillets. Cross-infection tests made with pools of homologous phages indicated that only 8 strains were resistant to all the phages. Some similarities between marine and terrestrial pseudomonads were found, as well as significant differences between species previously considered to be closely related.

Ploidy differences in Sporobolomyces salmonicolor and Candida albicans.

Abstract: Presumed diploid and haploid cultures ofSporobolomyces salmonicolor andCandida albicans were analysed for their DNA content per cell. Ratios of approximately 2 ∶ 1 were obtained by relating the DNA content of the two phases to ploidy.

The occurrence of thermophilic cellulolytic fungi in a pasture land soil

Abstract: Previous studies on the isolation of thermophilic fungi from soil have indicated their relative rarity. In the present investigation, however, many of the confirmed thermophilic fungi have been isolated from one pasture land, using soil enrichment methods and selective isolation media. These results point towards a wide distribution of thermophiles in soil and the cellulolytic activity of many of them.

Four new species ofPenicillium

Abstract: Descriptions with drawings are presented of four new species ofPenicillium: Penicillium fennelliae sp.n. Penicillium lignorum sp.n. Penicillium striatisporum sp.n. Penicillium hordei sp.n.

The rapid loss of viability of Azotobacter in aqueous solutions.

Abstract: When a low number ofAzotobacter vinelandii 12837 log phase vegetative cells (2 × 103 cells/ml) were removed from the culture liquid to water of the same temperature, a rapid loss of viability occurred depending on the procedure of washing and suspending. Death was not accompanied by visible lysis and the rate of loss of viability was less at lower temperatures, and in the presence of salts or cell-free filtrates from heavy cell suspensions in water. The die-off was erratic at increased cell concentrations and was accelerated by utilizable energy sources. Cells standing in a favorable ionic solution (0.1% NaCl) do not lose their viability while cells washed by a series of centrifugations with the same ionic solution show a progressive loss of viability with each washing. Phospholipids were found to leach from the cells into the aqueous solutions. Such cell death suggests instability of the cell membrane and the loss of osmotic or ionic control in the cells.

Elimination of mycoplasma from various cell cultures.

Abstract: Elimination ofMycoplasma orale-I from chronically infected cell lines was achieved either by treatment with a mixture of antibiotics in a hypotonic solution, or with 10 vol % of anti -M.orale rabbit serum in tissue culture medium. The latter treatment was preferable in most cases, as it was practically harmless to the cells. Inactivation of this antiserum had no effect on its potency. The antibiotic-hypotonic treatment was rather destructive, but to a different degree for the various cell cultures. Both methods were equally useful for the treatment of a monkey kidney cell line contaminated with a mycoplasma strain related toM.hyorhinis. The available anti -M.hyorhinis rabbit serum was toxic for the monkey cells when not inactivated. The potency of the antiserum was rather low and even lower after inactivation. However, prolonged treatment successfully eliminated the mycoplasma. Pre-incubation of the inactivated anti -M.hyorhinis serum with tissue culture medium to which 10% non-inactivated calf serum had been added, favoured the elimination of the mycoplasma. During the treatment of contaminated cell cultures with single antibiotics a strain related toM.hyorhinis became resistant to chlortetracyclin.M.orale- I was found to be resistant to various single antibiotics.

Unbalanced cell-wall synthesis in chloramphenicol-grown Rhodotorula glutinis.

Abstract: Rhodotorula glutinis grown in the presence of 500μg chloramphenicol per ml of medium has an increase in cell-wall material on a dry weight basis. This increase, from 13.4 to 24.5%, is closely paralleled by an increase in thickness as determined from electron micrographs of isolated cell walls. There is no detectable change in chemical composition of the walls. Vesicular elaborations of the plasmalemma were observed in the chloramphenicol-inhibited cells.

Systematics of the genera Saccharomyces, Schizosaccharomyces, Endomycopsis, Kluyveromyces, Schwanniomyces and Brettanomyces : Proton magnetic resonance spectra of the mannans and mannose-containing polysaccharides as an aid in classification

Abstract: The proton magnetic resonance spectra of the mannans and mannose-containing polysaccharides of 71 species ofSaccharomyces,Schizosaccharomyces, Endomycopsis, Kluyveromyces, Brettanomyces,Nematospora andSchwanniomyces and of some apparently related species ofTorulopsis were determined, grouped according to similarities in the spectra, and compared with those of the mannose-containing polysaccharides of some other yeasts. The spectra of the mannans produced bySaccharomyces, Kluyveromyces, Nematospora, Brettanomyces andTorulopsis were placed in 10 groups. The galactomannans formed by theSchizosaccharomyces species studied had very similar spectra which were placed in a single group. The spectra of the mannans of theSchwanniomyces species were likewise placed in a single group. The spectra of the mannans of theEndomycopsis species studied were not alike and were placed in three sub-groups.

Characterization of the oxidase-negative, gram-negative coccobacilli (the Achromobacter-acinetobacter group).

Abstract: More than 70 morphological, biochemical, and nutritional characters of 291 strains of aerobic, oxidase-negative, gram-negative coccobacilli representing theAchromobacter-Acinetobacter group of bacteria were examined. The subdivision of these bacteria is discussed in terms of glucolytic activity in a modified infusion medium, production of gelatinase, hemolysis of blood agar, growth on SS Agar, and utilization of selected organic compounds as the sole source of carbon and energy. On the basis of these characters the bacteria were divided into three major groups — glucolytic, nonglucolytic, and proteolytic which comprised six less clearly defined minor subgroups. Generic and species names currently applied to these bacteria are listed.

Two new yeast species isolated in Finland

Abstract: Two new yeast species,Sterigmatomyces elviae isolated from a male patient andTrichosporon fennicum isolated from a domestic cat, are described.

Isolation and identification of orsellinic acid and penicillic acid produced by Penicillium fennelliae Stolk.

Abstract: Fifteen days-old surface cultures ofPenicillium fennelliae Stolk grown on a modified Czapek medium were analyzed for the production of metabolites. The presence of orsellinic acid, orcinol and penicillic acid was proved by means of chromatographic and physicochemical methods.

Deoxyribonucleic acid base composition of species of Klebsiella, Azotobacter and Bacillus.

Abstract: Methods of molecular taxonomy were used to study the genome or deoxyribonucleic acid (DNA) of different strains of five genera of nitrogen-fixing bacteria. The DNA base compositions of all strains ofKlebsiella pneumoniae (previously classified as strains ofAchromobacter sp.,Aerobacter aerogenes orK. pneumoniae) occupy a fairly narrow range of values from 56.7 to 62.5% G-C content. No significant difference was observed in the DNA base composition of bacteria which can fix molecular nitrogen and that of strains which do not fix nitrogen.

On ultrastructures in Rhodopseudomonas gelatinosa and Rhodospirillum tenue.

Abstract: Tile photosynthetic pigments in bacteria are associated with membranous structures, named chromatophores (Pardee, Schachmann and Stanier, 1952).The morphology ofchromatophores is diverse: in Athiorhodaceae the occurrence of vesicular and several types of lamellar structures are described (Cohen-Bazire and Sistrom, 1966). Another type of chromatophore structure has now been demonstrated by us in Rhodopseudomonas gelatinosa (Molisch) van Niel and Rhodo,~pirillum tenue (Pfennig, personal communication). Cells grown anaerobically (witb 95 ~ N,. and 5 ~/o CO,,,) in the light contain tubular intrusions

Bacteriophages of psychrophilic pseudomonads. II. Host range of phage active againstPseudomonas putrefaciens

Abstract: Forty phages active against 4 terrestrial and against 10 of 24 marine strains ofP. putrefaciens were isolated from raw municipal sewage, fish-pier water, and refrigerated haddoch fillets. With one exception, phages active against marine strains were obtained only from pier water and fish fillets. Only two strains were attacked solely by their homologous phage while five of the marine strains were not attacked by any phage. Lytic reactions revealed no obvious differences between marine and terrestrial strains.

Serological analysis of eleven strains ofRhizobium japonicum

Abstract: The present communication reports a serological analysis of eleven strains ofRhizobium japonicum. The slow-diffusing thermostable antigens were found to be suitable for the basic differentiation of the somatic serogroups inRhizobium japonicum. One to three precipitation bands of the slow-diffusing thermostable antigens, one to two bands of the fast-diffusing thermostable antigens and one to three bands of the thermolabile antigens were detectable in the whole cell cultures ofR. japonicum by means of the immunodiffusion technique. Two basic somatic serogroups were differentiated on the basis of the slow-diffusing thermostable antigens. The thermolabile antigens were identical in most of the strains.