The serological grouping of Candida albicans strains isolated in the Kraków province in the years 1963–1965 by means of agar gel diffusion test

Summary One hundred forty sevenCandida albicans strains isolated from different clinical specimens were serologically differentiated. Agar gel diffusion test inBjörklund modification was applied inCandida albicans grouping. Autolysates prepared from the strains under study and from the group A and B standard strains ofHasenclever were used as antigens against anti-Candida albicans group A rabbit immune sera. Among 147 examined strains isolated from 107 patients, 137 strains were classified as belonging to group A and only 10 (9.3 %) strains as belonging to group B.     

Leclercia adecarboxylata Gen. Nov., Comb. Nov., formerly known asEscherichia adecarboxylata.

The nameLeclercia adecarboxylata is proposed for a group of the family Enterobacteriacae previously known asEscherichia adecarboxylata. Leclercia adecarboxylata can be phenotypically differentiated from all other species of Enterobacteriaceae. The members of this species are positive for motility, indole production, methyl red, growth in the presence of KCN, malonate, beta-galactosidase, beta-xylosidase, esculin hydrolysis, gas production fromd-glucose, and acid production fromd-cellobiose,d-lactose, melibiose,l-rhamnose, adonitol,d-arabitol, dulcitol, and salicin; the strains were negative for Voges-Proskauer, citrate (Simmons), H₂S (Kligler), lysine and ornithine decarboxylases, arginine dihydrolase, phenylalanine deaminase, gelatinase, DNase, Tween-80 hydrolysis, and acid production from myoinositol and alpha-methyl-d-glucoside. Fermentation ofd-raffinose,d-sucrose, andd-sorbitol is variable with strains. DNA relatedness of 11 strains ofL. adecarboxylata to three strains including the type strain of this species averaged 80% in reactions at 65°C. DNA relatedness to other species in Enterobacteriaceae was 2%–32%, indicating that this species was placed in a new genusLeclercia gen. nov. The type strain ofL. adecarboxylata is ATCC 23216.     

The rhizobia of lupinus densiflorus Benth., with some remarks on the classification of root nodule bacteria

Summary Four strains of rhizobia from Lupinus densiflorus Benth. were found to differ from the normal slow-growing strains of Rhizobium lupini by a rapid growth on agar medium, a somewhat different pattern of carbon metabolism, good growth in simple synthetic media, and also in their host plant relationships. Three strains had subpolar flagella like other lupine rhizobia, and the same was found to be predominant in a fourth strain previously described as having peritrichous flagellation.Two strains formed effectively nitrogen-fixing root nodules in Lotus corniculatus and Anthyllis vulneraria where the other two formed semieffective or ineffective nodules. All four strains formed ineffective nodules in Lotus uliginosus and Ornithopus sativus. The slow-growing strains of Rh. lupini mostly produce ineffective nodules in Lotus corniculatus but have now been seen to be effective in Lotus uliginosus.Instead of trying to define Rh. lupini as a cross-inoculation group it seems preferable to abandon it as a species and to transfer the fastgrowing strains to Rhizobium leguminosarum sensu Graham (1964) and De Ley and Rassel (1965), in spite of their predominantly subpolar flagellation. The familiar slow-growing strains would remain in the broad group of slow-growing root nodule bacteria with purely subpolar flagellation, called Phytomyxa japonica by Graham (1964) and Rhizobium japonicum by De Ley and Rassel (1965).     

Edwardsiella hoshinae,a new species of enterobacteriaceae

Eight strains isolated from birds, reptiles, and water constitute a new DNA hybridization group that is 37–58% related toEdwardsiella tarda and less than 10% related to other species of Enterobacteriaceae (SI nuclease method). This homogeneous group (78–100% relatedness within the group) constitutes a new species that is namedEdwardsiella hoshinae sp. nov. (type strain, CIP 78.56 ATCC 33379). Strains of this species produce acid fromd-mannitol, sucrose,d-trehalose, and salicin, and give a positive malonate test. Seven other strains that produced acid fromd-mannitol and sucrose (but not fromd-trehalose and salicin) and were malonate negative were found to belong to theEdwardsiella tarda DNA hybridization group. The base composition of the DNAs ofE. tarda andE. hoshinae is 55–58 mol% G+C.     

Two types ofPropionibacterium avidum with different isomers of diaminopimelic acid

Of 38 strains ofPropionibacterium avidum, most fell into one of two groups. One group (20 strains) hadLl-diaminopimelic acid as the diaminoacid of peptidoglycan, did not ferment inositol, and reacted with serum to strain VPI 0576; the other group (11 strains) hadDl-diaminopimelic acid as the peptidoglycan diaminoacid, fermented inositol, and reacted with serum to strain VPI 0670. DNA sequence similarity studies showed that, while overall intergroup similarity was about 80%, within each group the sequence similarities were over 90%. Seven strains were anomalous and did not fit exactly into either group.The results show that the isomer of diaminopimelic acid in peptidoglycan may differ in strains of a single species all of which show at least 80% DNA sequence similarity to each other.     

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.     

From scratch to value: engineering <Emphasis Type="Italic">Escherichia coli</Emphasis> wild type cells to the production of <Emphasis Type="SmallCaps">l</Emphasis>-phenylalanine and other fine chemicals derived from chorismate

Recombinant strains of Escherichia coli K-12 for the production of the three aromatic amino acids (l-phenylalanine, l-tryptophan, l-tyrosine) have been constructed. The largest demand is for l-phenylalanine (l-Phe), as it can be used as a building block for the low-calorie sweetener, aspartame. Besides l-Phe, an increasing number of shikimic acid pathway intermediates can be produced from appropriate E. coli mutants with blocks in this pathway. The last common intermediate, chorismate, in E. coli not only serves for production of aromatic amino acids but can also be used for high-titer production of non-aromatic compounds, e.g., cyclohexadiene-transdiols. In an approach to diversity-oriented metabolic engineering (metabolic grafting), platform strains with increased flux through the general aromatic pathway were created by suitable gene deletions, additions, or rearrangements. Examples for rational strain constructions for l-phenylalanine and chorismate derivatives are given with emphasis on genetic engineering. As a result, l-phenylalanine producers are available, which were derived through several defined steps from E. coli K-12 wild type. These mutant strains showed l-phenylalanine titers of up to 38 g/l of l-phenylalanine (and up to 45.5 g/l using in situ product recovery). Likewise, two cyclohexadiene-transdiols could be recovered.     

Researches on Leptospirosis ballum

Summary A case of an accidental human infection caused byLeptospira ballum, following a superficial scratch on a finger of the patient while handling a white mouse is described. This mouse proved to be a urinary carrier ofLeptospira ballum.Further investigations revealed that practically all mice of the same colony voidedLeptospira ballum with the urine.On examination of other mice-breeding colonies another colony infected withL. ballum was detected; in 1941 a strain which later also proved to beL. ballum, was isolated from the urine of a white mouse.The various strains were analysed; they were serologically identical toBorg Petersen's type strain although the virulence for guinea-pigs was much less.Mice from aL. ballum-positive colony proved to have no immunity to infection with virulentL. icterohaemorrhagiae, but — with one exception — remained alive and became carriers ofL. icterohaemorrhagiae, whilstL. ballum could not be detected any more in the urine of these mice.     

Immunochemical characterization of a polysaccharide antigen isolated from the oral microorganismEubacterium saburreum,strain 02/725

A serologically highly active polysaccharide antigen (PS 02/725) consisting of equimolar amounts ofd-glycero-d-galacto-heptose and a 6-deoxyheptose was isolated fromEubacterium saburreum, strain 02/725, by trypsin digestion and subsequent gel filtration and ion-exchange chromatography. Whole bacterial cells injected intravenously stimulated rabbits to produce precipitins and complement-binding antibodies of the IgG class specific for PS 02/725.     

Nodulation of actinorhizal plants byFrankia strains capable of both root hair infection and intercellular penetration

Summary A morphological analysis of the initiation and development of root nodules ofElaeagnus angustifolia andMyrica cerifera inoculated with pure-culturedFrankia strains DDB 011610 or DDB 020110 was undertaken. From ultrastructural observations it was determined that both of theseFrankia strains can infectElaeagnus by an intercellular penetration mechanism andMyrica by the root hair infection mechanism. This indicates that both of these strains have the ability to infect host plant roots by either of two mechanisms. The reverse, thatElaeagnus orMyrica could be infected by both mechanisms, was not observed. The infection and nodule development processes of these two plants in combination with these strains were similar to observations made in previous studies (Miller andBaker 1985,Torrey andCallaham 1979). However, one exception was identified in the development of the prenodule ofMyrica when infected with strain 011610, in that endophytic hyphae developed vesicles within the cells of the prenodule. This event has not been described before for any of the actinorhizal genera and may be an indication of less than optimal compatibility between the host plant and the symbiont.Contribution no. 876 of the Battelle-Kettering Laboratory.     

Metabolism of l-alanine in Desulfotomaculum ruminis and two marine Desulfovibrio strains

The degradation of l-alanine by three strains of sulfate-reducing bacteria that can grow with l-alanine as an energy source was investigated. In Desulfotomaculum ruminis and most likely also in two marine Desulfovibrio strains alanine is converted to pyruvate via an NAD-dependent alanine dehydrogenase. D. ruminis contained high activities of soluble NADH and NADPH dehydrogenases. In the marine strains the activities were much lower and the NADH dehydrogenase was partly associated with the membrane fraction.     

Paramoeba: A Common Marine Genus

Amoebae of the genus Paramoeba Schaudinn, 1896, have been found in British waters, further attesting to the wide distribution and common occurrence of these unusually interesting marine protozoa. The British strains belong to the species P. pemaquidensis Page, 1970, first described from the Atlantic coast of North America. The Nebenkörper appears to be characteristic of a morphologically and systematically homogeneous group of amoebae.     

Production of tyrosine from sucrose or glucose achieved by rapid genetic changes to phenylalanine-producing <Emphasis Type="Italic">Escherichia coli</Emphasis> strains

Escherichia coli K12 strains producing l-phenylalanine were converted to l-tyrosine-producing strains using a novel genetic method for gene replacement. We deleted a region of the E. coli K12 chromosome including the pheA gene encoding chorismate mutase/prephenate dehydratase, its leader peptide (pheL), and its promoter using a new polymerase chain reaction-based method that does not leave a chromosomal scar. For high level expression of tyrA, encoding chorismate mutase/prephenate dehydrogenase, its native promoter was replaced with the strong trc promoter. The linked ΔpheLA and Ptrc-tyrA::Kan^R genetic modifications were moved into l-phenylalanine producing strains by generalized transduction to convert l-phenylalanine-producing strains to l-tyrosine-producing strains. Moreover, introduction of a plasmid carrying genes responsible for sucrose degradation into these strains enabled l-tyrosine-production from sucrose.     

Glycoconjugates as possible receptors forStreptococcus pyogenes

The adherence capacities of M-protein-positive (M+) and M-protein-negative (M-) strains ofStreptococcus pyogenes were compared in human epithelial cells obtained from the pharynx (PEC) or from the buccal mucosa (BEC). Adherence to PEC was related to the presence of M protein (40.5±1.1 M+ and 17.8±0.6 M–S. pyogenes per PEC), whereas BEC showed adherence equally for M+ and M– strains. Different receptor sites may thus be involved on the two cell types. Preincubation of the bacteria with disialogangliosides (1 mg/ml), orN-acetylgalactosamine, ord-galactose (10 g/ml) resulted in diminished adherence of M+ strains to PEC but not to BEC. Chromatography ond-galactose-Sepharose 6B showed specific binding only of M+ group A streptococcal strains to gel beads. M– group A, and groups C and G streptococci did not bind. These observations suggest that the receptors on PEC for group A streptococci are distinct from those on BEC, and that most probably the attachment ofS. pyogenes to human pharyngeal cells occurs by specific, lectin-like binding to galactose residues on epithelial cells.     

Klebsiella ornithinolytica sp. nov., formerly known as ornithine-positiveKlebsiella oxytoca

The nameKlebsiella ornithinolytica sp. nov. is proposed for a group ofKlebsiella strains referred to previously as NIH Group 12 at the National Institute of Health, Tokyo. The members of this species are Gram-negative, encapsulated, nonmotile rods with the general characteristics of the familyEnterobacteriaceae and of the genusKlebsiella. They give positive results in tests for indole production, Voges-Proskauer, citrate utilization, lysine and ornithine decarboxylases, urease, -galactosidase, malonate utilization, growth in KCN, and esculin hydrolysis, and they produce acid and gas fromd-glucose, and acid froml-arabinose, cellobiose, lactose, melibiose, raffinose, rhamnose, sucrose, trehalose,d-xylose, adonitol,d-arabitol, myo-inositol, sorbitol, arbutin, salicin, -methyl-d-glucoside, and mucate. They give negative drolysis, DNase, pectinase, and acid production fromd-arabinose, melezitose, and dulcitol. They can grow at 4°C and 42°C, and do not produce any pigment. DNA relatedness of eight strains ofKlebsiella ornithinolytica to three strains including the type strain of this species averaged 88% in reaction at 75°C. DNA relatedness to the already recognizedKlebsiella species inEnterobacteriaceae was 1 to 20%. Phenotypic and DNA relatedness data also indicated that a group of organisms referred to as Enteric Group 47 orKlebsiella Group 47 at the Centers for Disease Control (Atlanta, Georgia) was identical withK. ornithinolytica. The type strain ofK. ornithinolytica is NIH 90-72 (JCM 6096).     

Regulation of homoserine dehydrogenase inAzotobacter species

The regulation of homoserine dehydrogenase activity was studied in nineAzotobacter strains belonging to five different species. In all the species the enzyme is subject to feedback inhibition byl-threonine andl-isoleucine, the first being much more active as inhibitor. The inhibition byl-threonine is noncompetitive with respect to NADPH and of mixed type with respect to aspartate-Β-semialdehyde; the inhibition byl-isoleucine is noncompetitive with respect to both substrates. The synthesis of homoserine dehydrogenase inAzotobacter chroococcum I.P. is somewhat repressed by 1mm l-methionine and 5mm l-isoleucine. In all the strains examined either NADPH or NADH can serve as cofactors for this activity, though the ratio of activity with the two pyridine nucleotides (NADPH/NADH) shows higher values (3.3–3.8) in the speciesmacrocytogenes andinsignis than in thechroococcum, beijerinckii andvinelandii group (1.5–1.6). The pattern of control of this enzyme in the genusAzotobacter is discussed in relation to other bacterial homoserine dehydrogenases. We are grateful to Dr. G. N. Cohen, Service de Physiologie Microbienne, Institut Pasteur, Paris, for helpful discussions and encouragements.     

Discrimination of Rhizobium japonicum,Rhizobium lupini,Rhizobium trifolii,Rhizobium leguminosarum and of bacteriods by uptake of 2-ketoglutaric acid,glutamic acid and phosphate

Rhizobium strains (one each of Rh.japonicum, Rh. lupini, Rh. leguminosarum) take up 2-ketoglutaric acid in general much faster and from lower concentrations in the medium than strains of Escherichia coli, Bacillus subtilis and Chromobacterium violaceum. A strain of Enterobacter aerogenes, however, is more similar to some Rhizobium strains. The same strains of Rhizobium take up also phosphate much faster and from lower concentrations than the other bacteria tested. 4 strains of Rh. lupini proved to be significantly different from 4 strains of Rh. trifolii in taking up l-glutamic acid from three to ten times lower concentration within 5 h. A similar difference was noticed between 5 strains of Rh. leguminosarum and 2 strains of Rh. japonicum for the uptake of 2-ketoglutaric acid and of l-glutamic acid. Isolated bacteriods from nodules of Glycine max var. Chippeway have a reduced uptake capacity for glutamic acid and for 2-ketoglutaric acid during the first 10–12 h, but reach the same value after 24 h as free living Rh. japonicum cells. The differences in the uptake kinetics are independent of cell concentration. The group II Rhizobium strains (Rh. japonicum and Rh. lupini, slow growing Rhizobium) are characterized by a rapid uptake of glutamic acid to a lowremaining concentration of 1–3×10^-7 M and an uptake of 2-ketoglutaric acid to a remaining concentration of 2–5×10^-7 M. The group I Rhizobium strains (Rh. trifolii and Rh. leguminosarum, fast growing Rhizobium), can be characterized by a much slower uptake of both substances with a more than ten times higher concentration of both metabolites remaining in the medium after the same time.     

Taxonomic studies on some gram-positive coryneform hydrogen bacteria

Recently isolated coryneform hydrogen bacteria were investigated under taxonomical aspects. Strains 7 C, RH 10, and 14 g are characterized by the snapping type of cell division, 68.5 to 69.7% GC content, dl-diaminopimelic acid in the cell wall, content of metachromatic granules, weak utilization of sugars and inhibitory effect of citrate. The strains are placed to the group 1—genus Corynebacterium—of the classification of coryneform bacteria of Yamada and Komagata (1972) and the name Corynebacterium autotrophicum sp.nov. is proposed.Strains 11 X and RH 12 are characterized by the bending type of cell division, a GC content of 70.2 and 70.5%, ll-diaminopimelic acid in the cell wall, absence of metachromatic granules, utilization of several sugars and no changes in cell morphology by citrate. The strains have to be placed to group 6 of coryneform bacteria.     

Determination of the Vi-antigen content of acetone-dried typhoid vaccines

Summary A number of typhoid strains and coli strain 5396/38 are compared for their Vi-antigen content with the aid of complement fixation and erythrocyte sensibilization tests. Acetone-killed and dried germs were used as source material. Parallel batches derived from the same strain showed good agreement. Moreover, results with erythrocyte sensibilization and complement fixation reaction were closely correlated. The conclusion is drawn that serological determination of the Vi-antigen of acetone-dried typhoid vaccine is possible. Further investigations, however, must be carried out to compare the functional capacity to produce antibodies of strains with the same serologically determined content of Vi-antigen. We thank Dr.M. Landy for the revision of this paper.