Differentiation of Coryne-bacterium diphtheriae of the mitis type found in diphtheria and ozaena. II. The rate of rapidity of glucose fermentation by C. diphtheriae type mitis and C. belfanti

Summary Eighty strains ofC. diphtheriae typemitis and 80C. belfanti strains were tested for the rate of rapidity of glucose fermentation according to the method ofParsons andFrobisher. 90% ofC. diphtheriae mitis strains, in contrast to only 13.7% ofC. belfanti strains, fermented glucose in 1 to 2 days. 76% ofC. belfanti strains fermented glucose in 3 to 4 days, whereas some strains needed 8 to 9 days to complete the fermentation. So the results of this test revealed next to that of nitrate reduction, a further difference between the strains ofC. diphtheriae typemitis found in diphtheria and ozaena.     

Metabolism ofl-proline toN-acetyl-d-glucosamine during germ tube formation ofCandida albicans

The metabolism ofl-proline toN-acetyl-d-glucosamine (GlcNAc) during germ tube formation ofCandida albicans (C. albicans) ATCC 1002 was studied. In uptake experiments, 6.9 nmol ofl-[¹⁴C]proline were taken up by 1×10⁶ cells during 3 h of incubation at 37°C. The percentage of germ tube formation was 94 under the same condition. The presence of GlcNAc reduced the uptake ofl-proline to 3.0 nmol. The percentage of germ tube formation was 95 in the presence and absence of GlcNAc. The [³H]GlcNAc uptake was 3.0 nmol and was constant whetherl-proline was present or not. After the preparation of a chitin fraction from germ tubes that were labeled withl-[¹⁴C]proline, the radioactivity froml-proline was detected in the glucosamine (GlcN) fraction by thin-layer chromatography (TLC). The metabolism ofl-proline to GlcNAc in chitin during germ tube formation was confirmed in this experiment.     

Delimitation and classification ofBromus fasciculatus (Poaceae)

The syntypes ofBromus madritensis var.delilei Boiss. comprise two different elements: specimens ofB. haussknechtii Boiss. and ofB. fasciculatus C. Presl s.l. By its lectotypification and on the basis of new morphological characters, the validation ofB. fasciculatus subsp.delilei (Boiss.)H. Scholz 1971 was substantiated. This subspecies represents the eastern marginal segregate of the south-Mediterranean species.B. fasciculatus var.alexandrinus Thell. is a minor variant of the typical subspecies.Dedicated to Prof.K. H. Rechinger on the occasion of his 80th birthday.     

Sur quelques hyménoptères intervenant dans la biocoenose de ravageurs de l’Artichaut dans le sud-est de la France

Summary No discovery of any parasite ofSphaeroderma rubidium Graell.,Ceratapion carduorum Kirbi, andDepressaria subpropinquella Stain. has been mentioned for France. However considerable collections ofMymaridae obtained on layings ofS. rubidium, show that parasitism may have a part in the limitation of this species. The few parasites of larvae and pupae ofC. carduorum obtained do not seem to have any considerable regulating influence. Although the parasitism of caterpillars ofD. subpropinquella byAngitia is frequent, the latter do not seem to be the essential factor in the evolution of the population of this species. The caterpillars ofChoreutis bjerkandrella thnb. may be used as a refuge by manyHymenoptera in summer, as a consequence of the scarcety ofMicrolepidoptera at a vulnerable stage.      

Influence des parasites oophages sur la régulation des populations deCassida deflorata Suff

Summary The layings ofCassida deflorata Suffr. are parasited in southern France byMymaridae (Anaphoidea sp. andFulmekiella ovata soyka),Trichogrammatidae (Monorthochaeta nigra Blood.) andEulophidae (Foersterella flavipes Foerst. andTetrastichus rhosaces Walk.). In the Alpes-Maritimes, two successive generations ofMymaridae andTrichogrammatidae seem to attack the layings ofC. deflorata. In Corsica, parasitism byMymaridae andF. flavipes is very frequent and may be the essential regulating factor of the populations ofCassida. In the other regions that have been studied, although the large number ofCassida imagos appearing in spring show that mortality during the aestivation and hibernation is not the primary factor, it does not seem that the limitation of the populations is essentially caused by the activity of oophagous insects.      

Studies on Aspergilli XVI. Effect of pH,temperature, and carbon and nitrogen interaction

Summary The effect of pH, temperature, and carbon and nitrogen interaction on the growth and sporulation ofAspergillus nidulans (Eidam)Wint.,A. rugulosus Thom &Raper,A. variecolor (Berk. &Br.)Thom &Raper andA. quadrilineatus was studied. All the moulds could grow on a wide range of pH (2.0 to 12.0) but the growth was poor on too acid and too alkaline media. Best growth ofA. rugulosus, A. quadrilineatus, andA. violaceus was seen at pH 6.5 and that ofA. nidulans andA. variecolor at pH 7.0. In general maximum production of perithecia was recorded between pH 6.0 and 8.0.All the above species ofAspergillus under study could grow between a temperature range of 10° C–48° C, but the growth was poor at 10° C and 48° C. The present moulds showed good growth at 20° C, 25°C, and 30° C. At 40° CA. nidulans andA. rugulosus showed moderate growth while the rest of the Aspergilli attained good growth. Temperatures between 20° C–30° C favoured excellent perithecial production.In general, little improvement in growth was noted on media containing good carbon and nitrogen sources. Malic acid was found to be useless when supplied singly. But, poor growth was recorded when supplied in combination with amino acids, amide, and peptone. This was due to the fact that these N sources also supplied carbon for their metabolism.     

Some interesting agarics from eastern Austria

Four very rare species ofAgaricales of dry grassland of the Galgenberg near Röschitz (Niederösterreich) are described and their taxonomy, affinities, ecology and distribution are discussed.Marasmiellus graminis (Murr.)Sing. & Hausknecht andCrinipellis subtomentosa (Peck.)Sing. var.mauretanica (Mre.)Sing. & Hausknecht are proposed as new combinations;Campanella inquilina Romagn. apudRedhead is a synonym ofC. caesia Romagn.     

Cultivation ofCandida blankii in simulated bagasse hemicellulose hydrolysate

Summary All fourCandida blankii isolates evaluated for growth in simulated bagasse hemicellulose hydrolysate utilized the sugars and acetic acid completely. The utilization ofd-xylose,l-arabinose and acetic acid were delayed by the presence ofd-glucose, but after glucose depletion the other carbon sources were utilized simultaneously. The maximum specific growth rate of 0.36 h^–1 and cell yield of 0.47 g cells/g carbon source assimilate compared with published results obtained withC. utilis. C. blankii appeared superior toC. utilis for biomass production from hemicellulose hydrolysate in that it utilizedl-arabinose and was capable of growth at higher temperatures.     

Substrate regulation of ascorbate transport activity in astrocytes

Astrocytes possess a concentrativel-ascorbate (vitamin C) uptake mechanism involving a Na⁺-dependentl-ascorbate transporter located in the plasma membrane. The present experiments examined the effects of deprivation and supplementation of extracellularl-ascorbate on the activity of this transport system. Initial rates ofl-ascorbate uptake were measured by incubating primary cultures of rat astrocytes withl-[¹⁴C]ascorbate for 1 min at 37°C. We observed that the apparent maximal rate of uptake (V _max) increased rapidly (<1 h) when cultured cells were deprived ofl-ascorbate. In contrast, there was no change in the apparent affinity of the transport system forl-[¹⁴C]ascorbate. The increase inV _max was reversed by addition ofl-ascorbate, but notD-isoascorbate, to the medium. The effects of external ascorbate on ascorbate transport activity were specific in that preincubation of cultures withl-ascorbate did not affect uptake of 2-deoxy-D-[³H(G)]glucose. We conclude that the astroglial ascorbate transport system is modulated by changes in substrate availability. Regulation of transport activity may play a role in intracellular ascorbate homeostasis by compensating for regional differences and temporal fluctuations in external ascorbate levels.     

Metabolic design in the amino-acid-producing bacteriumCorynebacterium glutamicum

The Gram-positive bacteriumCorynebacterium glutamicum is used for the industrial production of amino acids,e.g. ofl-glutamate andl-lysine. By cloning and expressing the various genes of thel-lysine pathway inC. glutamicum we could demonstrate that an increase of the flux ofl-4-aspartaldehydate tol-lysine could be obtained in strains with increased dihydro-dipicolinate synthase activity. Recently we detected that inC. glutamicum two pathways exist for the synthesis ofdl-2,6-diaminopimelate andl-lysine. Mutants defective in one pathway are still able to synthesize enoughl-lysine for growth but thel-lysine secretion is reduced to 50–70%. Using NMR-spectroscopy we could calculate how much of thel-lysine secreted into the medium is synthesizedvia the one and the other pathway. Amplification of the feedback-inhibition-insensitive-homoserine dehydrogenase and homoserine kinase in a highl-lysine-overproducing strain made it possible to channell of the carbon flow from the intermediate 4-aspartaldehydate toward homoserine, resulting in a high accumulation ofl-threonine. For a further flux froml-threonine tol-isoleucine the allosteric control of threonine dehydratase was eliminated. Dedicated to Dr. Z. Vaněk on the occasion of his 70th birthday Presented at theIUMS Congresses '94-7th International Congress of Bacteriology and Applied Microbiology Division, Prague, July 3–8, 1994 (Bacteriological Symposium BS-12Regulation of Microbial Product Overproduction).     

Alterations in the outer wall architecture caused by the inhibition of mycoside C biosynthesis inMycobacterium avium

Radiolabeled amino acids (l-U[C¹⁴]alanine,d-U[C¹⁴]alanine,l-U[C¹⁴]threonine, andl-U[C¹⁴]phenylalanine) were exponentially incorporated into the trichloroacetic acid (TCA)-insoluble material (whole cells) ofMycobacterium avium during the first 30–60 min of labeling. Bacteria labeled for 48 h were extracted with chloroform-methanol (21 vol/vol). The thin layer chromatography (TLC) analysis of native lipids showed that mycoside C was labeled by the amino acids used.d-cycloserine (d-CS) and other amino acid analogs were examined as potential inhibitors of mycoside C biosynthesis. It was found thatd-CS caused about 27% inhibition, whereaso-,p-, andm-fluoro-dl-phenylalanine (Fl-phe) caused 80%–90% inhibition of the mycoside C biosynthesis. Judging from the data on inhibition experiments, it was concluded that the mycoside C biosynthesis started from the fatty acyl end and proceeded by the stepwise addition ofd-phenylalanine,d-allo-threonine, andd-alanine. Thed-alanyl-d-alanine peptidoglycan intermediate did not seem to serve as a donor ofd-alanine for mycoside C biosynthesis. Ultrastructural observation of the bacteria treated withd-CS showed only partial alteration of the outer wall layer, whereasm-Fl-phe treatment caused profound alterations. Successive transfers of the bacteria in growth medium supplemented withm-Fl-phe resulted in extensive disorganization of the outer layer.     

Peptidoglycantyp und Zusammensetzung der Zellwandpolysaccharide vonCellulomonas cartalyticum und einigen coryneformen Organismen

Cellulomonas cartalyticum was found to contain a peptidoglycan type different from that of the other species ofCellulomonas. The diamino acid is lysine instead of ornithine and the interpeptide bridge consists ofd-Asp-d-Ser. The same peptidoglycan type occurs inCorynebacterium manihot, Brevibacterium liticum andArthrobacter luteus. These non cellulolytic organisms are most likely not closely related withCellulomonas cartalyticum, as indicated by the very different G+C content of their DNA, although they formed a narrow cluster includingC. cartalyticum when numeric taxonomical methods were applied.

     

The pathway ofD-cycloserine biosynthesis inStreptomyces garyphalus

Incubation experiments using washed cells and toluene treated cells ofStreptomyces garyphalus showed that O-acetyl-L-serine and hydroxyurea are intermediates in the biosynthesis ofD-cycloserine. The formation of [¹⁴C]O-ureidoserine from O-acetyl-L-serine and hydroxyurea was demonstrated by incubating an enzyme solution with¹⁴C-labelled substrates. Desalted cell-free extract catalyzed the conversion of O-ureido-D-serine toD-cycloserine in a reaction requiring ATP and Mg²⁺. The results suggested the following pathway forD-cycloserine biosynthesis.     

Nutritional studies on species and mutants of Lepista,Cantharellus, Pleurotus and Volvariella

The purpose of this study is to determine the ability of select Agaricales species to utilize various sources of carbon, nitrogen, vitamins, and growth hormones. Fungi selected for the studies include:Cantharellus clavatus Fries,C. cibarius Fries,Lepista nuda (Bull. ex. Fries)Cooke,Pleurotus ostreatus (Jacq. ex. Fries)Kummer, andVolvariella volvacea (Bull. ex. Fries)Singer. Three strains ofC. cibarius and one mutant ofV. volvacea (V135), V134, were employed to determine if nutritional requirement differences occurred. One species,V. volvacea, is grown commercially as a cottage industry in the Orient (Alicbusan &Ela, 1961) while the other species currently have no commercial value. All species studied possess pleasing flavors and have potential use in the mushroom production industry.A literature compilation of the nutritional regulation of basidiocarp formation and vegetative growth of Agaricales was made with specific mention to the named species (Volz &Beneke, 1969). Recent nutritional studies with one or more of the specific species include those byYusef &Allam (1967), andEger (1970).     

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.     

An optimum environment for the culturing of cytospora isolates from stone fruits. I. Temperature

Summary Four isolates ofCytospora cincta Fr. and 2 ofC. leucostoma Fr. were obtained from diseased Italian prune, President plum and Bing cherry trees.The minimum temperature for growth of these fungi was found to be 3° C. Temperatures of 45 °C. were lethal to all cultures. The optimum temperature for theC. cincta isolates on solid and liquid media was found to be 30° C.; for theC. leucostoma isolates, nearly 25° C. OneC. cincta isolate produced greatest radial growth on the solid medium at 35° C., but in the liquid medium produced maximum mycelium at 30° C.All factors considered, the conclusion was reached that the best single temperature for laboratory culture of the fungi was 30° C.Approved by the Director of the Idaho Agricultural Experiment Station as Research Paper No. 493.     

Deacetylation ofN-acetylthienamycin to thienamycin by a cell-free extract ofStreptomyces cattleya,the thienamycin producer

Summary A cell-free extract from the thienamycin producer,Streptomyces cattleya, has been found to deacetylate the co-product,N-acetylthienamycin. The pH optimum of the reaction is 7.5. Due to the lability ofN-acetylthienamycin, we used thed andl forms of the synthetic substrateN-chloroacetylvaline. We found that the enzyme is anl-deacetylase, has a molecular weight of 58 000, is stable up to 40°C, acts optimally at 45°C, is stable at pH 5–8, is not activated by divalent metal ions and is inhibited by Hg⁺⁺, Cu⁺⁺ andp-chloromercuribenzoate. This is the first report of an extract from a carbapenem producer which carries out the deacetylation ofN-acetylthienamycin, suggesting that the acetylated derivative is a precursor of thienamycin.Abbreviations THM thienamycin - N-AcTHM N-acetylthienamycin - CFE cell-free extract - N-Cl-Ac-l-Val N-chloroacetyl-l-valine - N-Cl-Ac-d-Val N-chloroacetyl-d-valine     

Catabolism of arginine byCarnobacterium spp. isolated from vacuum-packed sugar-salted fish

The ability ofCarnobacterium spp. originally isolated from vacuum-packed, sugar-salted fish to catabolize arginine was examined. All strains were able to produce citrulline, ornithine, and NH₃ from arginine, presumably by the arginine deiminase pathway. The metabolism of arginine was concurrent with acid production from glucose for one strain ofCarnobacterium sp. but delayed for one strain ofCarnobacterium piscicola. The arginine catabolism was not inhibited in the presence of 2% glucose for three strains of carnobacteria during growth in test broth and/or shrimp extract. Growth as well as arginine catabolism was delayed for two strains of carnobacteria by lowering the temperature from 9°C to 4°C. A similar result was obtained by incubating one strain ofC. piscicola in CO₂. None of the compoundsl-citrulline,l-ornithine hydrochloride, and (NH₄)₂SO₄ had any effect on growth or arginine catabolism of this strain. Neither did pH of the medium affect the time for initiation of arginine catabolism.     

Heterostyly in species ofNarcissus (Amaryllidaceae) andHugonia (Linaceae) and other disputed cases

The hypothesis ofHenriques andFernandes that several Iberian species ofNarcissus (Amaryllidaceae) are tristylous is reconsidered. Contrary to the opinion ofBateman and most subsequent authors, we believe that the available evidence indicates that some populations ofN. triandrus andN. fernandesii, at least, are tristylous; other populations ofN. triandrus are distylous.Hugonia cf.penicillanthemum (Linaceae) from new Caledonia is distylous, but it remains possible that other species ofHugonia are tristylous. The disputed occurrence of heterostyly in S. African species ofBauhinia (Leguminosae),Cleome (Capparaceae) andAneilema (Commelinaceae), and inAgelaea (Connaraceae) is discussed.     

Parasitisme multiple chezDiparopsis watersi roths. (Lep. Agrotidae)

Summary Larvae ofDiparopsis watersi roths, are parasitized in the Tchad Republic by an unspecifiedMermitidae and byCarcelia evolans wied. (Tachinidae). TheMermis is only active during the rainy season, when 40% ofD. watersi larvae are commonly parasitized. More than one individual are able to develop inside the same host, a maximum of 44 being quoted; yet, solitaryMermis are of a greater length, the biggest recorded being 325 mm long. C. evolans is found all along the active life ofD. watersi. There is usually only one dipterous larva in each host: Out of more than a hundred cases, only twice two and thrice threeTachinidae were encountered in the same host. Evolution of theseC. evolans larvae is uncertain, only one of them developping into an adult. Association ofMermis andC. evolans, though exceptional, was also observed. This was described eight times through a three years survey; again, only one of the dipterous larvae developped into adult. The most extraordinary case reported concerns threeC. evolans encountered together with two Nematoda in the same host. An explanation to the unusual multiparasitism observed may be sought in an exceptional abundance ofC. evolans, combined with a relative scarcity of the host during the month of september 1956.