- Volume 14, Issue 3, 1956
Volume 14, Issue 3, 1956
- Obituary
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- Article
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The Nature of the Lesion in the Succinate-requiring Mutants of Neurospora crassa: Interaction between Carbohydrate and Nitrogen Metabolism
More LessSUMMARY: Mutants of Neurospora crassa requiring dicarboxylic acids for an immediate growth response (suc and at suc) oxidize acetate, and are inhibited by fluoroacetate with consequent citric acid accumulation to approximately the same extent as the wild-type. The concentration of nitrogen (as ammonium and nitrate salts) present in the conventional growth medium is inhibitory to the growth of these mutants and leads to an accumulation of aeetylmethylcarbinol, pyruvic acid and α-ketoisovaleric acid. This inhibition is reduced and growth is stimulated by the addition of dicarboxylic acids or by diminution of the nitrogen present in ‘minimal’ medium. The addition of nitrogen salts to suc mutants probably diverts dicarboxylic acids (already in short supply) from the catalysis of the oxidation of C2 fragments via the tricarboxylic acid cycle to other reactions. This effect of nitrogen salts upsets the already precarious dicarboxylic acid balance of the suc mutants leading to a growth requirement and to the accumulation of intermediates.
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A Proposed Revision of the Genus Pullularia
More LessSUMMARY: Morphological and physiological studies were made of 12 strains of the genus Pullularia. Although inability to ferment carbohydrates has been reported characteristic of this genus, 10 of the strains produced acid from one or more carbohydrates. It is proposed that the genus be emended to include the new species P. fermentans.
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Some Observations on the Antigenic Structure of Trichloroacetic Acid Extracts in Escherichia and the Paracolon Group
More LessSUMMARY: All the somatic antigens of ‘ smooth ’ type of some Escherichia and paracolon strains studied are not extracted by trichloroacetic acid (TCA), though they may be by boiling or by mechanical disruption of the cell.
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Cytological Effects of Ultraviolet Radiation and Azaserine on Corynebacterium diphtheriae
J. C. Davis and S. MuddSUMMARY: Ultraviolet irradiation and the radiomimetic antibiotic substance L-azaserine have similar effects in promoting the formation of radially enlarged cells and elongated filaments, and in the accumulation of metaphosphate in metachromatic granules, with a strain of Corynebacterium diphtheriae. Dosages effective for the formation of enlarged cells and filaments are lower than for stimulation of metaphosphate accumulation.
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Spontaneous Mutation in Stored Spores of a Streptomyces sp.
More LessSUMMARY: Mutations were induced by storing non-dividing asexual spores of a Streptomyces sp. at 4° for varying periods of time. Certain stages among spores germinated at 27° were more ‘ sensitive ’ than others to this mutagenic action. This high sensitivity was not correlated with particular nuclear stages or with nuclear number. The increased frequency of mutants cannot be attributed to selection (either decreased survival of the wild type or selective growth of previously existing mutants), to increased background (gamma) radiation or to temperature shock. The data indicate that mutation rate is a function of the physiological condition of the spore and that mutations may occur in the absence of nuclear division.
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Cytochrome c 3 and Desulphoviridin; Piǵments of the Anaerobe Desulphovibrio desulphuricans
More LessSUMMARY: Suspensions of various mesophilic strains of Desulphovibrio desulphuricans show absorption bands attributable to a cytochrome and a green protein; there are small differences in the position of absorption maxima depending on the strain and culture medium. Both pigments have been extracted, together with flavins rich in flavinadenine dinucleotide; an electrophoretically and chromatographically pure preparation of the cytochrome has been obtained and is designated c 3. The green protein has been termed ‘desulphoviridin’.
Cytochrome c 3 is a soluble autoxidizible thermostable haemoprotein (reduced bands at 553, 525 and 419 mμ.) of low redox potential ( − 205 mV.), high iso-electric point (pH >10) and containing 0·9 %Fe. Degradation studies indicate that it is a bifunctional haemato-haematin with the thio-ether haem-apoprotein links also found in cytochromes c and f; its m.w. is approx. 13,000 (S20,w = 1·93 × 10−13). Spectroscopic data for various derivatives including haemin c 3 and a porphyrin derivative are recorded. Material purified to at least 94 % by cellulose and ion-exchange chromatography acts as carrier in the reduction in hydrogen of sulphite, thiosulphate, tetra-thionate or dithionite by detergent-treated bacterial preparations; a similar role has been demonstrated with cell-free systems which reduce sulphite, thiosulphate and tetrathionate. Benzylviologen would replace cytochrome c 3. No preparation has been obtained showing c 3-linked sulphate reduction; the evidence for this depends on difference spectra and competition by known sulphate antagonists.
Oxidation of H2 or organic compounds with O2 has been demonstrated with these bacteria; the H2/O2 reaction takes place fastest in an atmosphere containing 4 % O2, when oxygen is frequently reduced faster than sulphate. The reaction requires the mediation of cytochrome c 3 and is probably a consequence of the autoxidizibility of c 3.
Desulphoviridin is a thermolabile, soluble, acidic porphyroprotein absorbing at 630, 585 and 411 mμ.; no metabolic function has been detected. It is stable over a limited pH range and decomposes readily, yielding a chromophoric group which fluoresces red in ultraviolet light, absorbs at 595 mμ. in neutral and alkaline solution (solution red) and at 612 mμ. in acid solution (solution blue-green). This material can be purified by chromatography on ‘Florisil’ or paper. It is very photo-sensitive and water-soluble. Its character is obscure; it may be a highly carboxylated chlorin. Spectroscopic data are recorded.
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The Transfer of Defective Lambda Lysoǵeny between Strains of Escherichia coli
More LessSUMMARY: The infective transfer of defective lambda lysogeny from a defective prophage to sensitive Escherichia coli has been observed. The agent of transfer appears to be lambda phage in which has been incorporated the hereditary defective element. It is concluded that the existence of the phage genes responsible for defective lysogeny is not limited to the prophage condition.
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The Chemical Composition of the Cell Wall in some Gram-positive Bacteria and its Possible Value as a Taxonomic Character
More LessSUMMARY: Hydrolysates of cell-wall preparations of more than 60 strains of corynebacteria, lactobacilli, streptococci, staphylococci and other Gram-positive cocci have been examined by paper chromatography. A very high proportion of the amino acid moiety of the cell-wall complex could in each case be accounted for in terms of 3 or 4 of the amino acids alanine, glutamic acid, lysine, diaminopimelic acid, aspartic acid and glycine. These were associated with varying combinations of sugars and amino sugars. In general, each bacterial genus appears to have a characteristic pattern of cell-wall components, particularly in regard to the amino acids present. Variations in the relative proportions of the sugars appear to differentiate the individual species within a genus. The possible value of cell-wall composition as a taxonomic character is discussed.
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The Continuous Culture of Bacteria; a Theoretical and Experimental Study
More LessSUMMARY: A theoretical treatment of continuous culture is given, which allows quantitative prediction of the steady-state concentrations of bacteria and substrate in the culture, and how these may be expected to vary with change of medium, concentration and flow-rate. The layout and operation of a small pilot plant for the continuous culture of bacteria are described. This plant has been operated continuously for periods of up to 4 months without breakdown or contamination of the culture. No alterations in the properties of the organisms studied have occurred during such periods of continuous culture. Results are given of a series of experiments on the continuous culture of Aerobacter cloacae in a chemically defined medium, designed to allow quantitative comparison with the results predicted by the theory. The relative advantages of batch and continuous culture as production processes are discussed, and it is concluded that continuous culture may usually be expected to show a five to tenfold increase in output as compared with a batch process.
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Influenza Virus Nucleic Acid: Relationship between Biological Characteristics of the Virus Particle and Properties of the Nucleic Acid
More LessSUMMARY: Five A strains and three B strains of influenza virus were purified and found to contain ribonucleic acid in amounts varying from 0·75 to 1·1 %. The proportion of the purine and pyrimidine derivatives in the nucleic acid of each strain was determined. When the ratio adenine + uracil : guanine + cytosine was evaluated, the following values were obtained: A strains —PR 8, 1·27 ± 0·02; MEL, 1·22 ± 0·01; WSE, 1·26 0±01; SWINE, 1·24 ± 0·04; CAM, 1·28 ± 0·01. B strains—LEE, 1·42 ± 0·04; MIL, 1·43 ± 0·05; ROB, 1·38 ± 0·01. The nucleic acid content of PR8 virus preparations of varying degrees of incompleteness was determined both by specific absorption at 260 mµ. and by estimation of phosphorus present in the nucleic acid extract. As measured by both methods, virus preparations of low infectivity were found to have a decreased nucleic acid content. The demonstration of specific differences in the nucleic acid of A and B strains and of the relationship between the infectivity and nucleic acid content of the virus affords strong evidence that the nucleic acid is an intrinsic part of the influenza virus particle.
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Sedimentation Behaviour and Electron Microscopic Examination of Purified Influenza Virus
More LessPurified preparations of influenza virus were examined in the ultracentrifuge and electron microscope principally to assess the degree of purification achieved. As indicated in the main paper (Ada & Perry, 1956) the same purification procedure was used for each batch of virus.
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Dihydrostreptomycin and Anaerobiosis-Comparison with Other Antibiotics and its Selectivity with regard to Obligate Anaerobes
More LessSUMMARY: The sensitivity to various antibiotics of certain facultatively anaerobic bacteria when growing under aerobic or anaerobic conditions was compared. The antibiotics tested were: dihydrostreptomycin, penicillin, chloramphenicol, erythromycin, carbomycin, chlortetracycline, oxytetracycline, tetracycline. Dihydrostreptomycin was unique amongst the antibiotics tested in being less active against facultative anaerobes when they were growing anaerobically than when growing aerobically. Dihydrostreptomyein was relatively inactive against Clostridium welchii and C. novyi, and its action was governed by the inoculum size. Dihydrostreptomyein is not a useful selective agent for the isolation of C. welchii.
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A Study of the Factors Influencing Non-genetic Variation in a Strain of Fusarium oxysporum
More LessSUMMARY: The colour variation of a strain of Fusarium oxysporum Schlecht ex Fr. was found to depend on the production of carotenoids and naphthoquinones. The carotenoids were only produced as a result of exposure to light, whereas the production of naphthoquinones, which were pH indicators, was mainly regulated by the carbon/nitrogen ratio of the medium. The morphogenetic effects of radiation and of carbon/nitrogen ratio were also considered. A low carbon/nitrogen ratio favoured chlamydospore formation and aerial mycelium, whereas ultraviolet radiation, and to a lesser extent light, promoted macrospore and sclerotium production.
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The Influence of certain Micro-organisms on the Formation of Perithecia by Chaetomium globosum
More LessSUMMARY: The localized formation of perithecia in colonies of Chaetomium globosum growing in association with certain other micro-organisms (e.g. Aspergillus fumigatus) is associated with the presence of organic phosphates, especially phospho-glyceric acid, in the medium. These phosphates are normal metabolic products of the associated micro-organism, and only when they diffuse into the medium and locally reach a critical concentration does the fruiting response follow.
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Automatic Control of pH Value in Cultures of Micro-organisms
More LessSUMMARY: Equipment for the automatic control of pH value in cultures of micro-organisms is described. The apparatus was designed for a 2 l. scale continuous culture apparatus. The equipment will continuously control pH value with an accuracy of ± 0·05 unit for periods of many hundred hours. The pH value may be changed at will by turning a knob on the controller. It is sufficient to check the pH meter standardization once every day or two. The main components of the apparatus are standard commercial products.
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The Gonidial Stages in Spirillum spp. and Vibrio spp
More LessSUMMARY: In twelve strains, comprising three species of saprophytic Spirillum and two of Vibrio, a life cycle was observed in which small gonidia, resembling very tiny bacteria, sometimes motile by means of polar flagella, were formed within enlarged mother-cells. After a period of reproduction in this phase, the gonidia became progressively enlarged to reconstitute the original form. In one species of Spirillum, the cycle was induced by the addition of penicillin to the medium, and in this and certain other respects was reminiscent of the L-cycle.
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A New Vi-phage Type of Salmonella typhiwith a Discussion of Methods of Preparation of Typing Phages for New Vi-Types
More LessSUMMARY: A new Vi-phage type of Salmonella typhi, T4904, is described. The homologous typing phage (phage 4904) was prepared from phage D 6, a host-range mutant of Vi-phage II. The new type belongs to the E group and consists of type E1 carrying the determining phage d6. The formula El(d6) has therefore been assigned to it. The host range of phage 4904, the homologous typing phage, is considered in relation to the structural formula of T4904. This formula fully explains the ability of the phage to lyse strains other than T4904. The general aspects of adaptation of Vi-phage II to new Vi-types of S. typhi are discussed. It is shown that the older attitudes to this subject require modification in the light of recent work. The applications of the results of study of the temperate type-determining phages to the solution of practical problems in Vi-phage typing are described.
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A Study of the Metabolism of Phenolic Compounds by Soil Fungi Using Spore Suspensions
More LessSUMMARY: Spore suspensions were used to investigate the metabolism of p-hydroxy- benzaldehyde, ferulic acid, syringaldehyde and vanillin by Haplographium sp., Hormodendrum sp., Penicillium sp. and Spicaria sp. The intermediate products of their metabolism, p-hydroxybenzoic, syringic and vanillic acids, were found to be attacked by adaptive enzymes. The formation of these enzymes was greatly decreased by the antibiotic citrinin.
The results of experiments, based on a technique in which fungal spore suspensions were used in place of hyphae, are presented in this paper. Under investigation was the metabolism of certain phenolic compounds related to lignin, which have already been reported to be utilized by soil microfungi as sources of carbon (Henderson & Farmer, 1955).
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A Modified Method for Preparing and Counting Viable Suspensions of Tubercle Bacilli
More LessSUMMARY: A modified method derived from that of Fenner, Martin & Pierce (1949) is described for the preparation of homogeneous suspensions of single organisms of one strain of Mycobacterium tuberculosis at concentrations of about 108 single organisms per ml. A technique is also described for the accurate assessment of viable counts in a period of 14 days.
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Some Mutational Changes in the Photosynthetic Pigment System of Rhodopseudomonas spheroides
More LessSUMMARY: Some 50 mutants, which differ from the wild type in the nature of their pigment systems, were isolated from the non-sulphur purple bacterium, Rhodopseudomonas spheroides. They fall into five main groups. The group of colourless mutants is incapable of photosynthetic growth, and devoid both of chlorophyll and carotenoids. The remaining groups still contain bacteriochlorophyll, but differ chemically from the wild type and from one another in their carotenoid pigments; all are capable of photosynthetic growth. The wild type contains two principal carotenoids, one red and one yellow. The dark red mutants contain both wild-type carotenoids and traces of a newr red carotenoid. The brown mutants contain approximately equal amounts of the wild-type yellow carotenoid and of neurosporene, together with traces of the wild-type red carotenoid. The green mutants contain neurosporene and a dihydroxy derivative, but neither of the wild-type pigments. The blue-green mutants contain phytoene, a colourless polyene, but are completely devoid of coloured carotenoids. The infra-red spectrum of cells of the blue-green mutant is markedly different from that of the wild type and of the other photosynthetic mutants, despite the fact that its chlorophyll is chemically identical with theirs.
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Dictyostelium polycephalum n.sp.: a New Cellular Slime Mould with Coremiform Fructifications
More LessSUMMARY: A unique species of Dictyostelium is described for which the binomial Dictyostelium polycephalum is proposed because of the seemingly branched character of its fructifications. This slime mould has been isolated repeatedly from samples of surface soil and decomposing leaves collected from deciduous forests in various parts of the United States. As in other members of the Acrasieae, its vegetative phase consists in the independent growth of free-living myxamoebae which feed upon bacterial cells, and its fruiting phase is characterized by the inflowing of these cells to form multicellular organizations preparatory to fructification. D. polycephalum differs from previously known species of the genus, particularly for its formation of cell aggregates which give rise to varying numbers of long, thin migrating pseudo- plasmodia, and for the potential capacity of each of these to form subsequently a small coremiform fructification consisting of multiple sorocarps. Whereas some simple sorocarps are regularly produced, fructifications under optimal conditions typically consist of from two to ten adherent sorocarps. In such coremiform structures, the sorophores of individual sorocarps, although clearly distinguishable microscopically, are tightly appressed for approximately three-quarters of their length, at which level they diverge sharply and at their apices bear globose spore heads, or sori. The spores are elliptical to reniform as in most species, but germinate by the swelling and dissolution of the spore wall in a, median plane rather than by longitudinal splitting.
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Criteria for Establishinǵ the Validity of in vitro Studies with Rumen Micro-organisms in so-called Artificial Rumen Systems
More LessSUMMARY: Several criteria of normal rumen function which can be applied to in vitro studies with the whole rumen microbial population are suggested. These include: the maintenance of numbers and normal appearance of the bacteria, selenomonads and protozoa of the rumen; the maintenance of normal rates of digestion of cellulose, starch and protein, and of normal interactions between these; the ability to predict quantitative results in vivo. An ‘artificial rumen’ was constructed, consisting of a cellophan sac containing rumen liquor and substrate dialysing against a complex mineral solution whose composition was based on that found in rumen liquor, the whole being incubated at 39° in an atmosphere of nitrogen and carbon dioxide. This system was shown to meet the criteria which are suggested, with reasonable success for periods of about 8 hr.; over longer periods an increasing failure to meet the biological criteria was seen. For the microbial population to remain normal in numbers and activity it was shown to be necessary to use as test substrate in vitro only substances similar to the diet fed to the animal from which the rumen liquor inoculum was taken.
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Proteolysis by Rumen Micro-orǵanisms
More LessSUMMARY: Toluene-treated washed suspensions of rumen bacteria break down proteins largely to amino acids; in the absence of toluene bacterial deaminases are active. Unlike the deaminases, the presence of proteases does not depend, to any great extent, on the presence of readily attacked protein in the diet of the host animal. Extracts of acetone-dried powders of the bacteria also show proteolytic activity. Rumen protozoa are also proteolytic, and ammonia appears to be the end product of their nitrogen metabolism. Ammonia production due to the protozoa is not as sensitive to toluene as is the case with bacteria. Much of the ammonia production in the rumen in the absence of substrate appears to be due to the endogenous metabolism of the protozoa. Extracts of acetone powders, and extracts prepared by simple freezing and thawing of the protozoa, contain active proteases.
In an artificial rumen apparatus it was shown that when digestion was complete, about half the N and C of added casein could be recovered as ammonia and volatile fatty acids respectively. Most of the remainder could not be accounted for analytically, and was presumed to be used for microbial growth, which had occurred. When starch or some other polysaccharides were added to the artificial rumen apparatus as well as casein, the production of ammonia was lowered. This was shown not to be due to any effect on proteolysis or deamination, and was presumed to be due to the increased utilization for microbial growth of some breakdown product of casein.
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Reversion of a Pleuropneumonia-like Organism to a Corynebacterium during Tissue Culture Passage
More LessSUMMARY: A strain of a pleuropneumonia-like organism (PPLO) isolated from urethral exudate from a case of non-specific urethritis was studied in HeLa cell tissue cultures. Although the organisms entered the cell cytoplasm, they did not produce marked damage or proliferate luxuriantly until filtrate from a broth culture of Staphylococcus pyogenes or yeast extract was added to the infected tissue cultures. The organisms subsequently isolated from tissue cultures initially inoculated with PPLO and yeast extract showed conversion from PPLO form to L form of growth. Further culture of the L form, especially with the aid of mucin, resulted in conversion of the L form to a corynebacterium. This corynebacterium was indistinguishable culturally, biochemically and serologically from a corynebacterium isolated on rabbit blood agar plates which had been inoculated with a portion of the original urethral exudate from the same case of non-specific urethritis. The view is expressed that other human genital strains regarded at present as PPLO may be found to be L forms of Corynebacterium spp. The criteria for identification of PPLO and L forms are discussed.
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