- Volume 109, Issue 1, 1978
Volume 109, Issue 1, 1978
- Obituary
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- Biochemistry
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Further Observations on Carbohydrate Metabolism and its Regulation in Azotobacter beijerinckii
More LessCertain enzymes of glucose catabolism in Azotobacter beijerinckii were studied and their activities in steady-state chemostat cultures were measured under various nutrient limitations. 2-Keto-3-deoxy-6-phosphogluconate aldolase was separated from 6-phosphogluconate dehydratase by affinity chromatography and the previously observed inhibition of the Entner-Doudoroff enzymes by tricarboxylic acids and ATP was attributed to chelation of Mn2+ and Mg2+ which activate the dehydratase. Glucose-6-phosphate dehydrogenase was unaffected by phosphoenolpyruvate while fructose-1,6-bisphosphate aldolase was activated by Co2+, K+ or NH4 + ions. Transketolase, transaldolase and triosephosphate isomerase were present but previous reports of 6-phosphogluconate dehydrogenase activity were shown to be artefacts. The findings confirm the major role of the Entner-Doudoroff pathway in glucose catabolism in A. beijerinckii. Pyruvate dehydrogenase, a key enzyme for carbon entry to the tricarboxylic acid cycle and to poly-β-hydroxybutyrate synthesis, was inhibited by acetyl-coenzyme A and NADH.
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Anaerobic Transport of Serine and 2-Aminoisobutyric Acid by Staphylococcus epidermidis
More LessA membrane-bound ATPase detected in extracts of anaerobically grown Staphylococcus epidermidis was inhibited by a variety of compounds which inhibit ATPases in other organisms. Serine and 2-aminoisobutyric acid (AIB) were shown to enter the organism via the same transport system. The transport of AIB, the membrane potential and the transmembrane pH gradient were partially or completely abolished by the same inhibitors and also by uncoupling agents and lipid-soluble ions. It is proposed therefore that this ATPase generates and maintains an electrochemical gradient of protons across the cytoplasmic membrane of S. epidermidis capable of driving AIB uptake. Studies of AIB-induced proton movements suggested that AIB enters via a proton symport mechanism.
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- Development And Structure
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The capsules of Corynebacterium equi and Streptococcus equi
More LessThe capsules of Corynebacterium equi and Streptococcus equi were examined by electron microscopy after staining with ruthenium red. They were compared with the capsule of Klebsiella pneumoniae which had previously been examined using the same procedure ( Springer & Roth, 1973 ). The capsule of C. equi had a laminated appearance. When S. equi was grown on solid medium, its capsule appeared as radially arranged projections capped by a thick electron dense layer. When grown in liquid medium, S. equi produced a capsule which showed as short thick projections with no layer external to them.
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- Genetics And Molecular Biology
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Genetics of Radiation Sensitivity in the Slime Mould Dictyostelium discoideum
More LessMutations leading to increased sensitivity to 60Co gamma rays and/or ultraviolet light in the slime mould Dictyostelium discoideum have been assigned to eight loci on the basis of linkage and complementation tests. The linkages of these loci are: radA, linkage group I; radB, linkage group VI; radC, linkage group III; radD, linkage group IV; radF, linkage group IV; radG, linkage group III; radH, linkage group II; and radI, linkage group III. The radC mutants, which were isolated on the basis of increased sensitivity to ultraviolet light, are as resistant as their parental strains to 60Co gamma rays, whereas all other mutants studied are sensitive to both ultraviolet light and 60Co gamma rays. A new temperature-sensitive mutation, tsgL606, has been assigned to linkage group I.
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Thermosensitive Antibiotic Resistance Plasmids in Enterobacteria
More LessOf 775 conjugative plasmids found in enterobacteria mediating antibiotic resistance, 24 (3·1%) were thermosensitive (ts); they were most common in Klebsiella pneumoniae. Ts plasmids were also found in all the samples of sewage and river water examined. Over half of 73 ts plasmids from unrelated sources mediated resistance to chloramphenicol in addition to several other antibiotics. Many of them mediated resistance to mercury (53·4%), arsenite (38·4%) and tellurite (79·5%) but not to copper, cobalt and silver. Fifty-eight belonged to incompatibility group H2 and 12 belonged to the H1 group. Resistance to mercury, arsenite and tellurite was common in strains containing H2 plasmids but not in H1 plasmids. The 73 plasmids transferred at high rates at 22 and 28°C and at lower rates at 15°C; they transferred at very low rates or not at all at 37°C. They could be divided into two sets according to whether they transferred at a high or at a low rate at 33°C. Unlike the prototype plasmid, Rts 1, they were solely or mainly ts for transfer and not for replication and only one of them brought about a marked reduction in growth rate of its host organism at 42°C. None of the 73 plasmids mediated colicin or haemolysin production. Three plasmids, all from K. pneumoniae, mediated utilization of lactose, two of sucrose and raffinose and three, all belonging to group H1, of citrate. None of the plasmids increased the pathogenicity of Salmonella typhimurium for chicks or Escherichia coli k12 for mice.
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Arsenic Resistance in Enterobacteria : its Transmission by Conjugation and by Phage
More LessA high proportion of strains of Escherichia coli (599 of 716 strains), Shigella (15 of 18), Proteus (32 of 33) and Klebsiella pneumoniae (42 of 61), but not of Salmonella (15 of 209), were resistant to sodium arsenite; the incidence of arsenite resistance was higher in animal than in human strains of E. coli. Transmissible arsenite resistance, which was mainly plasmid-borne, was common in resistant strains of K. pneumoniae (22 of 34 tested) and Salmonella (11 of 12), but not in resistant strains of E. coli (10 of 98), Proteus (0 of 32) or Shigella (0 of 6). In three K. pneumoniae strains from which the arsenite resistance genes could not be transferred by direct methods, they were mobilized by implanting the conjugative plasmids F or I intothem. In two S. typhimurium strains with transmissible arsenite resistance, the resistance genes were located in the genome of the phage with which they were lysogenized. In several of the enterobacterial strains the plasmid-borne arsenite resistance was not associated with antibiotic resistance or any other character known to be transmissible. In many of the K. pneumoniae strains, though, it was often associated with transmissible antibiotic, mercury and tellurite resistance and especially with transmissible lactose utilization. The available evidence strongly suggested that the arsenite resistance genes were located on the Lac plasmid. The conjugative plasmids in several of the K. pneumoniae strains were temperature-sensitive. Many of the arsenite-sensitive K. pneumoniae strains grew on culture media containing mixtures of sodium arsenite and sodium nalidixate at concentrations of each which by itself would not permit their growth. Arsenite-resistant K. pneumoniae cultures did not grow under such conditions but any arsenite-sensitive mutants they contained did.
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Lipopolysaccharide Core Defects in Salmonella typhimurium Mutants Which Are Resistant to Felix O Phage but Retain Smooth Character
More LessFOR mutants of Salmonella typhimurium are resistant to Felix O phage, whose receptor includes the N-acetylglucosamine branch of the lipopolysaccharide (LPS) core, but smooth in cultural properties, antigenic character and phage sensitivity pattern ( MacPhee et al., 1975 ). The rfa(FOR) genes determining the FOR character of nine mutants were transduced into a smooth cysEpyrE recipient: the nine FOR transductants (and a tenth FOR mutant) were then made rfb (i.e. unable to make O chains) by transduction or Hfr crosses. The rfb FOR strains were sensitive to FO phage but nearly all of them showed a somewhat reduced efficiency of plating and diminished rate of adsorption of the phage. This observation and the Ra (complete core) serological activity of their LPS (tested by haemagglutination inhibition) indicate the presence of some, but less than the normal number of, completed core chains in FOR rfb LPS. On the basis of the sensitivities of the FOR transductants and their rfb derivatives to various “rough-specific” phages, their increased sensitivities to some antibiotics and to deoxycholate and the serological activity of the rfb FOR LPS in various incomplete core systems, the mutants were divided into three groups: (i) five mutants with probable defects in previously undetected rfa gene(s) concerned with formation of both the galactose I and the galactose II units of the LPS core; (ii) two mutants with defects inferred to affect the structure of the inner part of the core and also interfere with addition of the N-acetylglucosamine branch; (iii) three mutants in which no type of incomplete core could be detected, probably affected in formation of the inner part of the core chain. The mutation of one mutant of the last class, unlike those of the other nine mutants tested, lay outside the cysE–pyrE segment, in the 90 to 116 min region of the linkage map.
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The Relevance of a Study of a Temperature-sensitive Ballooning Mutant of Aspergillus nidulans Defective in Mannose Metabolism to our Understanding of Mannose as a Wall Component and Carbon/Energy Source
More LessA temperature-sensitive mutant strain of Aspergillus nidulans has been isolated which failed to grow normally on minimal agar medium at 43 °C unless mannose was supplied as a sole carbon source. The mutant has been given the symbol mnrA455 for mannose relief. Under restrictive conditions (43 °C) the mutant produced extensive areas of swollen hyphae, called balloons, and wall preparations from cultures grown at 40 °C had approximately one-third of the mannose found in walls of control cultures. The mnrA455 strain produced a more thermolabile phosphomannose mutase than the wild-type strain, so it is suggested that the mutation is in the structural gene coding for this enzyme.
The mnrA455 strain failed to grow on minimal agar medium containing 0·1% (w/v) mannose and 0·9 % (w /v) glucose at 43 °C. This property was used to obtain revertants, one of which was a double mutant mnrA455 manA1. Growth of the double mutant in media containing [14C]mannose at 43 °C showed that 84 % of the label found in the wall occurred in mannose in comparison with 33% in the wild-type control. Autoradiography of cultures of the double mutant using [3H]mannose showed predominant incorporation of label into the tip region of growing hyphae. A manA1 strain, isolated following haploidization of a diploid, was unable to grow on media containing glucose or mannose alone. The manA1 strain had reduced phosphomannose isomerase (EC 5.3.1.8) activity. The manA1 mutation, which is epistatic to mnrA455, has been located to linkage group VIII and the mnrA455 mutation to linkage group V. A pathway is presented for the utilization of mannose in A. nidulans.
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- Medical Microbiology
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The Effect of Pneumonia Induced in Mice with Mycoplasma pulmonis on Resistance to Subsequent Bacterial Infection and the Effect of a Respiratory Infection with Sendai Virus on the Resistance of Mice to Mycoplasma pulmonis
More LessThe effect of pneumonia induced by Mycoplasma pulmonis in mice on the resistance of the lung to additional bacterial infection was examined. The effect of pneumonia induced by Sendai virus on the resistance of mice to M. pulmonis was also investigated and compared with the effect of Sendai virus on resistance to Staphylococcus aureus. Sendai virus infection decreased subsequent resistance to M. pulmonis in proportion to the virus dose. Decreased resistance to subsequent S. aureus and M. pulmonis infection was greatest at about the same time after inoculation of virus and was related to virus-induced lesions. Besides affecting the resistance of mice to subsequent mycoplasma infection, Sendai virus could enhance an existing mycoplasma infection. Pneumonia induced by M. pulmonis did not decrease resistance to subsequent bacterial infection. The mechanism whereby Sendai virus decreases host resistance is therefore similar for bacteria and mycoplasmas, but pneumonia induced by mycoplasmas does not have the same effect.
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- Physiology And Growth
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Attenuation by Divalent Cations of the Effect of the Phytoalexin Rishitin on Erwinia carotovora var. atroseptica
More LessRishitin at 300 μg ml–1 rapidly decreased oxygen uptake by Erwinia carotovora var. atroseptica suspended in 0·1% (w/v) peptone water. Variation in the composition of the suspending medium affected sensitivity to rishitin, with Mg2+ and, to a lesser extent, Ca2+ decreasing bacterial sensitivity. Addition of 100 μg rishitin ml–1 affected cell membrane permeability causing an increase in conductivity of the suspending medium. Inhibition of oxygen uptake by the cationic surfactant hyamine 2389 or by sodium lauryl sulphate was also alleviated by the addition of Mg2+ suggesting that rishitin may act directly on cell membranes of bacteria, possibly in a manner similar to a cationic surfactant or a membrane-active antibiotic. The results also suggest that the sensitivity of E. atroseptica to rishitin in potato tubers may be affected by variation in their Mg2+ or Ca2+ content. Phaseollin did not inhibit respiration of E. atroseptica.
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Regulation of Enzyme Synthesis in the Arginine Biosynthetic Pathway of Pseudomonas aeruginosa
More LessIn Pseudomonas aeruginosa the synthesis of only two out of eight arginine biosynthetic enzymes tested was regulated. Comparisons were made between the specific activities of these enzymes in bacteria grown on arginine or on its precursor, glutamate. N 2-Acetylornithine 5-aminotransferase (ACOAT), an enzyme involved in both the biosynthesis and catabolism of arginine, was induced about 14-fold during growth of the organism on arginine as the only carbon and nitrogen source, and the anabolic ornithine carbamoyltransferase (aOTC), a strictly biosynthetic enzyme, was repressed 18-fold. Addition of various carbon sources to the arginine medium led to repression of ACOAT and to derepression of aOTC. Fructose, which supported only slow growth of P. aeruginosa, had a weak regulatory effect on the synthesis of the two arginine enzymes while citrate, a good carbon source for this organism, had a strong effect. The repression of ACOAT by citrate was not relieved by adding cyclic AMP to the medium. Under a variety of growth conditions leading to different enzyme activities, a linear relationship between the reciprocal of the specific activity of ACOAT and the specific activity of aOTC was observed. This inverse regulation of the formation of the two enzymes suggested that a single regulatory system governs their synthesis. Such a view was supported by the isolation of citrate-resistant regulatory mutants which constitutively formed ACOAT at the induced level and aOTC at the repressed level.
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‘Viscotaxis’, a New Behavioural Response of Leptospira interrogans (biflexa) Strain b16
More LessWhen in a non-viscous environment and confronted with a viscous one, Leptospira interrogans (biflexa) strain b16 preferentially selected the latter. We have designated this positive response to a viscosity gradient as ‘viscotaxis’. Using an originally designed experimental chamber, a pool of leptospires were faced with capillary tubes containing either polyvinylpyrrolidone (PVP) or N-2-hydroxyethylpiperazine-N′-2-ethanesulphonic acid (HEPES). Leptospires suspended in HEPES responded positively to 2% (w/v) PVP in capillaries, and migrated into them in large numbers in 1 h. No response was observed when the chamber and capillary tubes contained solutions of the same viscosity. As the viscosity of PVP was increased, a proportionally larger number of leptospires migrated into it. This newly observed aspect of leptospiral behaviour may have ecological significance.
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Analysis of DNA Content, Nitrogenase Activity and In Vivo Protein Synthesis of Rhizobium leguminosarum Bacteroids on Sucrose Gradients
More LessBacteroids of Rhizobium leguminosarum (strain PRE) isolated from pea root nodules (Pisum sativum) were separated by sucrose density gradient centrifugation, according to their stage of development from bacteria to mature bacteroids. The DNA content per cell, as measured by cytofluorometry, increased with development. Nitrogenase components in soluble bacteroid proteins were present in highest concentrations in mature bacteroids. The ratio of the amounts of Mo–Fe protein to Fe protein was not constant in different stages of development. Incorporation of 35SO4 2– into soluble bacteroid proteins in the nodule was maximal in the youngest stages. In mature bacteroids nitrogenase was synthesized preferentially although less 35SO4 2– was incorporated into total soluble protein. Nitrogenase activity, measured as ATP- and S2O4 2–-dependent acetylene reduction by EDTA/toluene treated bacteroids, was high in mature bacteroids and low in the youngest stages.
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Regulation of Extracellular Collagenase Production in Achrumobacter iuphagus
More LessAchromobacter iophagus synthesized extracellular collagenase in a highly aerated peptone medium at the late-exponential and early-stationary phases of growth. Collagenase synthesis was subject to end-product repression and was repressed by various amino acids and ammonium ions. Glutamine caused severe repression of collagenase production. Collagenase synthesis was sensitive to catabolite repression by glucose and a number of carbon sources. Cyclic AMP, dibutyryl cyclic AMP and cyclic GMP did not relieve catabolite repression. Glucose and 2-deoxyglucose caused a severe transient repression. Although rifampicin and chloramphenicol immediately inhibited RNA and protein synthesis, respectively, they failed to inhibit collagenase production completely. No intracellular preformed collagenase was detected and collagenase production ceased when induced cells were washed and resuspended in buffer.
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Fusion of Yeast Protoplasts Induced by Polyethylene Glycol
More LessProtoplasts, prepared from auxotrophic strains of Saccharomyces cerevisiae, Schizosac-charomyces pombe and Hansenula wingei, were mixed to give intraspecific complementary combinations. Polyethylene glycol (PEG) was added to induce agglutination and fusion. Some of the fused products grew on the surface of solid minimal medium forming large vacuolated bodies. Others reverted to hybrid cells when embedded in solid minimal regeneration medium. The cytological and preliminary genetical analyses suggest a syn-karyon formation and integration of genetic markers from parental strains. The frequency of intrageneric fusions assessed from the number of protoplasts growing on the surface of minimal agar was estimated to be 1 to 3 %, while the frequency of hybrid colony formation in regeneration medium was less than 1 %.
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- Short Communication
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- Taxonomy
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Chemical and Numerical Taxonomy of Strains Received as Gordona aurantiaca
More LessEight strains of Gordona aurantiaca, 89 representatives of the genera Mycobacterium, Nocardia and Rhodococcus and a single strain of Brevibacterium linens were the subject of numerical phenetic analyses using 92 unit characters. The G. aurantiaca cluster was robust in different numerical analyses and equivalent in rank to the genera Mycobacterium, Nocardia and Rhodococcus. The presence of a characteristic mycolic acid and completely unsaturated menaquinones distinguished G. aurantiaca strains from mycobacteria, nocardiae and rhodococci. The congruence between the numerical and phenetic data indicate that G. aurantiaca strains form a distinct taxon resembling other mycolic acid-containing taxa in wall, fatty acid and polar lipid composition. Since the genus Gordona is no longer valid, a new niche is needed for G. aurantiaca.
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Some Bits and Pieces of the Genus Nocardia: N. carnea, N. vaccinii, N. transvalensis, N. orientalis and N. aerocolonigenes
More LessSince 1951 when our taxonomic study of the genus Nocardia began, only eight strains of N. carnea (Rossi-Doria) Castellani & Chalmers, four strains of N. vaccinii Demaree & Smith, five of N. transvalensis Pijper & Pullinger, 21 of N. orientalis (Pittenger & Brigham) Pridham and 14 of N. aerocolonigenes (Shinobu & Kawato) Pridham have been found. These five groups of strains are described and are compared with strains of nine accepted species of Nocardia. With the possible exception of N. orientalis, more strains of these five taxa must be assembled and examined to establish the reliability of our descriptions and identifications.
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Isolation and Peptidoglycan of Gram-negative Hydrocarbon-utilizing Thermophilic Bacteria
More LessFour Gram-negative, non-sporulating, aerobic, obligate thermophilic bacteria, isolated from non-thermal environments by enrichment with n-heptadecane as substrate, utilized n-alkanes, carbohydrates and organic acids as sole source of carbon and energy and also grew on complex media. The growth rate of these organisms, when utilizing n-heptadecane as substrate, was markedly increased by adding a low concentration (7·5 mg l–1) of yeast extract. They grew optimally between 55 and 65 °C, and at a pH between 6·2 and 7·5. The mol% G + C for all was between 51 and 58. On the basis of the amino acid and amino sugar compositions of their peptidoglycan, these organisms and other Gram-negative thermophilic bacteria can be divided into four distinct groups. Group A includes the newly isolated hydrocarbon-utilizing bacteria which have nearly equimolar amounts of glutamic acid, alanine, diaminopimelic acid and glucosamine. Group B consists of obligate hydrocarbon-utilizing microbes that have lower molar ratios of glutamic acid and diaminopimelic acid, and contain either ornithine or lysine. The previously isolated non-hydrocarbon-utilizing thermophiles (k-2, Thermus aquaticus yt-1, Thermus x-1) and a newly isolated organism from a hot spring comprise group C and contain glycine, ornithine, no diaminopimelic acid, and much lower molar ratios of glutamic and muramic acids than in groups A and B. Thermomicrobium roseum lacked peptidoglycan and is placed separately in group D.
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Volumes and issues
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Volume 170 (2024)
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