Composition and biological properties of lipopolysaccharides isolated from Schizothrix calcicola (Ag.) Gomont (Cyanobacteria).

The most common cyanobacterium contaminating drinking water systems in southwestern Pennsylvania is Schizothrix calcicola. Lipoplysaccharides (LPS) were isolated from this species by hot phenol-water extraction. The polysaccharide moiety was composed of glucosamine, galactose, glucose, mannose, xylose and rhamnose. The lipid A part contained beta-hydroxylauric, myristic, pentadecanoic, palmitic, beta-hydroxypalmitic, stearic, oleic, and linoleic acids. In contrast to many LPS isolated from Enterobacteriaceae, the dominant component was not beta-hydroxymyristic but beta-hydroxypalmitic acid. The LPS induced Limulus lysate gelation and Schwartzman reaction but was nontoxic to mice. The identity of LPS was verified by alkali and lysozyme treatment. The results suggest that S. calcicola is one of the principal sources of endotoxins in water systems using open finished-water reservoirs.     

Composition, structure, and biological properties of lipopolysaccharides from different strains of Pseudomonas syringae pv. atrofaciens

The composition, structure, and certain biological properties of lipopolysaccharides (LPS) isolated from six strains of bacteria Pseudomonas syringae pv. atrofaciens pathogenic for grain-crops (wheat, rye) are presented. The LPS-protein complexes were isolated by a sparing procedure (extraction from microbial cells with a weak salt solution). They reacted with the homologous O sera and contained one to three antigenic determinants. Against the cells of warm-blooded animals (mice, humans) they exhibited the biological activity typical of endotoxins (stimulation of cytokine production, mitogenetic activity, etc.). The LCD of the biovar type strain was highly toxic to mice sensitized with D-galactosamine. The structural components of LPS macromolecules obtained by mild acidic degradation were characterized: lipid A, core oligosaccharide, and O-specific polysaccharide (OPS). Fatty acids 3-HO-C10:0, C12:0, 2-HO-C12:0, 3-HO-C12:0, C16:0, C16:1, C18:0, and C18:1 were identified in lipid A of all the strains, as well as the components of the hydrophilic part: glucosamine (GlcN), ethanolamine (EtN), phosphate, and phosphoethanolamine (EtN-P). In the core LPS, glucose (Glc), rhamnose (Rha), L-glycero-D-manno-heptose (Hep), GlcN, galactosamine (GalN), 2-keto-3-deoxy-D-mannooctonic acid (KDO), alanine (Ala), and phosphate were present. The O chain of all the strains consisted of repeated elements containing a linear chain of three to four L- (two strains) or D-Rha (four strains) residues supplemented with a single residue of 3-acetamido-3,6-dideoxy-D-galactose (D-Fucp3Nac), N-acetyl-D-glucosamine (D-GlcpNAc), D-fucose (D-Fucf), or D-Rhap (strain-dependent) as a side substitute. In different strains the substitution position for Rha residues in the repeated components of the major rhamnan chain was also different. One strain exhibited a unique type of O-chain heterogeneity. Immunochemical investigation of the LPS antigenic properties revealed the absence of close serological relations between the strains of one pathovar; this finding correlates with the differences in their OPS structure. Resemblance between the investigated strains and other P. syringae strains with similar LPS structures was revealed. The results of LPS analysis indicate the absence of correlation between the OPS structure and the pathovar affiliation of the strains.     

Immunobiological properties of lipopolysaccharides isolated from Fusobacterium nucleatum and F. necrophorum

Lipopolysaccharides (LPSs) were isolated from Fusobacterium nucleatum ATCC 10953 and F. necrophorum ATCC 25286 by the hot phenol/water procedure. F. nucleatum LPS was composed of 16% (w/w) carbohydrate, 10% (w/w) hexosamine and 40% (w/w) fatty acid, while F. necrophorum LPS was composed of 26% (w/w) carbohydrate, 12% (w/w) hexosamine and 28% (w/w) fatty acid. These LPS preparations induced mitogenic responses in spleen cells of BALB/c, BALB/c (nu/nu) and C3H/HeN mice, and these responses were suppressed by the addition of polymyxin B. The preparations also induced the polyclonal responses of C3H/HeN spleen cells. In addition, enhanced glucose utilization and interleukin-1 production by murine peritoneal macrophages were demonstrated. Neither spleen cells nor macrophages from the 'LPS-nonresponsive' C3H/HeJ mouse were activated by LPSs from the Fusobacterium species.     

Structural characteristics and biological properties of Pseudomonas fluorescens lipopolysaccharides

The structure and biological properties of lipopolysaccharides (LPSs) from strains IMB 4125 (=ATCC 13525) and IMB 7769 of the bacterium Pseudomonas fluorescens (biovar I) were studied in vitro. LPSs were similar in the composition of lipid A and the core lipid but differed in the structure of O-specific polysaccharide chains, which was corroborated by the absence of serological relationships between them. The toxicity (LD50) of LPSs of P. fluorescens with respect to D-glucosamine-sensitized mice was 40-50 times lower than the toxicity of the classic endotoxins, LPSs of E. coli. The LPSs studied stimulated the production of tumor necrosis factor (TNF) and nitric oxide (NO) by mouse peritoneal macrophages. The rates of TNF and NO synthesis induced by the LPSs of interest were eight to nine and three to five times lower, respectively, than the corresponding parameters of the control LPSs of E. coli 055:B5 and 026:B6. Additionally, LPS preparations of the P. fluorescens strains induced TNF synthesis by monocytes of human whole-blood preparations. Certain differences in biological properties of these strains have been revealed, which could be due to the characteristic features of LPS structure and composition in different cultures.     

Composition of O-antigenic lipopolysaccharides from Enterobacter cloacae

Analyses have been carried out on lipopolysaccharides (LPS) from 14 strains of Enterobacter cloacae representing different O serotypes. All of the products appeared to have a composition and architecture typical of enterobacterial LPS, but points of interest include the absence of phosphate residues from the core oligosaccharide, the presence of both L-glycero-D-mannoheptose and D-glycero-D-mannoheptose (ratio usually about 4:1), and the presence in lipid A of small amounts of fatty acids with odd numbers of carbon atoms (mainly C13) in addition to tetradecanoic acid and 3-hydroxytetradecanoic acid. Monosaccharides identified as components of polymeric fractions from the LPS were glucose, galactose, mannose, rhamnose, glucosamine, galactosamine, fucosamine, and galacturonic acid. Most polymeric fractions also probably contained an O-acetyl substituent. Closely similar chemotypes found for the polymeric fractions from the LPS of cross-reacting serotypes support the view that these fractions contain the O-antigenic determinants and represent the side chains of the LPS.     

Partial purification and some properties of urease from the alkaliphilic cyanobacteriumNostoc calcicola

Urease extracted from an alkaliphilic diazotrophic cyanobacteriumNostoc calcicola was partially purified and some of its properties were studied. Urease purified 39-fold from the crude enzyme extract showed its optimum activity at pH 7.5 and at 40°C with aK _m value of 120 μmol/L. The enzyme was found to be sensitive to metal cations, particularly Hg²⁺, Ag⁺ and Cu²⁺. 4-Hydroxymercuribenzoate (a mercapto-group inhibitor) and acetohydroxamic acid (a chelating agent of nickel) inhibited, the enzyme activity completely. These results suggest the involvement of an SH-group and Ni²⁺ in the activity of urease fromN. calcicola.     

Chemical and biological properties of a peptido-glucan fraction from Armillaria mellea (Basidiomycetes).

A peptide-glucan fraction from Armillaria mellea (Basidiomycetes) was isolated and some aspects of its chemical structure were determined. The glucan is linked to the peptide portion which represents 30% w/w of the complex. Treatment with alkali destroys most of the threonine and leads to the separation of the peptide and carbohydrate moieties indicating the involvement of the hydroxyl group of threonine in the peptide-glucan linkage. The results of partial hydrolysis, methylation studies and Smith degradation involving periodate oxidation, borohydride reduction and acid hydrolysis indicate that the polysaccharide moiety consists of beta(1 leads to 3) and beta(1 leads to 6)-linked D-glucose residues. This peptide-rich glucan fraction showed a significant antitumor activity.     

Chemical and biological properties of mycobactins isolated from various mycobacteria

Nine different strains of mycobacteria grown on media deficient in iron all produced mycobactins. Most strains produced one mycobactin in great preponderance. Mycobacteria from clearly distinct taxonomic groups gave mycobactins differing in the structure of their nuclei. One group of taxonomically related mycobacteria produced mycobactins having the same nucleus but with different distributions of side chains within the homologous mixtures. Simple methods are described for identifying mycobactins on a small scale; these may be of value in classifying mycobacteria. Structures are proposed for mycobactin A from Mycobacterium aurum, mycobactin R from M. terrae, mycobactin F, produced together with mycobactin H by M. fortuitum, and mycobactins M and N from M. marinum. The first three of these differ from known mycobactins in details of substitution and configuration of asymmetric centres in the nucleus. Mycobactins M and N are substantially different, having only small acyl groups (acetyl and propionyl respectively) at the hydroxamic acid centre of the mycobactic acid moiety. Both are homologous mixtures having long-chain saturated 3-hydroxy-2-methyl acid fragments in the cobactin moiety. All mycobactins so far isolated promote almost maximal growth of M. johnei at 30ng./ml. in liquid medium. The activity of some mycobactins extends to much lower concentrations, mycobactin S showing significant growth promotion at 0.3ng./ml. Mycobactin M or N in combination with mycobactins having a long side chain in the mycobactic acid moiety exerts a mutually antagonistic effect on the growth of M. johnei, the mixture giving less growth than either mycobactin separately. Mycobactin M also decreases the growth of M. kansasii and M. tuberculosis on liquid media. These antagonistic effects are probably caused by a lengthening of the lag phase.     

Immunochemical characteristics and serologic properties of lipopolysaccharides isolated from various Brucella species using various extraction methods

The results of the comparative analysis of LPS isolated by different methods of extraction from the cultures of several Brucella species differing in their virulence are presented. Purified LPS preparations have been obtained from Brucella virulent and vaccine strains by using such methods as water-phenol extraction, Boivin's method, mild alkaline hydrolysis of the antigen according to Boivin's procedure. The presence of certain relationship between the method used for the extraction of Brucella LPS to be compared and their chemical composition, immunological characteristics and serological activity has been established. As shown in this investigation, in the process of the preparation of a highly sensitive diagnosticum for the passive hemagglutination test the use of LPS obtained from Brucella virulent strains, but not from the vaccine strain, by the method of mild alkaline hydrolysis according to Boivin's procedure is expedient. The data presented in this work indicate that the soluble complex of lipid A obtained from Brucella LPS has been found to possess serological activity. The results of the study of the serological properties of lipid A indicate that the lipid component may also play a certain role in the manifestation of the serological activity of Brucella LPS.     

Biological properties of lipopolysaccharides from Bordetella species

Biological activities of lipopolysaccharides (LPS) extracted from Bordetella pertussis, B. parapertussis and B. bronchiseptica were compared with those of Escherichia coli LPS. The LPS preparations from B. pertussis showed biological activities comparable to those of E. coli LPS in terms of lethal toxicity in galactosamine-sensitized mice, pyrogenicity in rabbits, mitogenicity in C3H/He spleen cell cultures, macrophage activation, and induction of tumour necrosis factor. All the activities of LPS preparations from B. parapertussis, except mitogenicity, were lower than those of E. coli LPS. LPS from B. parapertussis gave the greatest mitogenic action of all those tested. Biological activities stronger than or comparable to those of E. coli LPS were observed for LPS from B. bronchiseptica.     

Removal of Pb2+ from aqueous system by live Oscillatoria laete-virens (Crouan and Crouan) Gomont isolated from industrial effluents

Cyanobacteria have been found to be potential biosorbents of metal ions from waste water. The Pb(2+) removal capacity of growing cells of indigenous cyanobacterium Oscillatoria laete-virens (Crouan and Crouan) Gomont was studied under batch experiments and it was found capable of removing Pb(2+) of lower concentrations (below 100?mg L(-1)). The effects of different concentrations of Pb(2+), on the growth rate of alga were also evaluated. The research parameters include the pH of the solution, contact time, initial concentration of Pb(2+), and culture density. Of the parameters studied, the pH of the solution was found to be the most crucial. The removal of Pb(2+) peaked at an initial pH of 5. The data obtained from the equilibrium experiments were found well fitting with the Langmuir isotherm with a maximum sorptive capacity (q (max)) of 20.36?mg?g(-1), indicating a good biosorbtive potential of growing cells. This was confirmed using scanning electron microscope and energy dispersive X-ray analysis, which showed the adsorption of lead on the surface of the cell. The species could tolerate a concentration as high as 60?mg L(-1) of Pb(2+). It was observed that the removal obeyed the pseudo-second-order kinetics. The percentage removal was found to decrease with increasing metal concentration, from 10 to 100?mg L(-1). FTIR analysis indicates the involvement of amino, carboxylic and amide groups in the sorption process. Among the desorbing agents evaluated, an efficient recovery of 90.2?% was achieved by HCl, in 24?h. Thus Oscillatoria laete-virens (Crouan and Crouan) Gomont seems to be a promising metal biosorbent for the treatment of Pb(2+), in waste waters.     

Antitumor activity and other biological properties of two phenotypes isolated from BCG

Summary Two phenotypes isolated from the BCG strain of Mycobacterium tuberculosis were found to differ from each other and from the parental strain by their chemical and antigenic compositions and by some of their biological properties. When used as immunotherapeutic agents in admixture with cancerous cells, one of these phenotypes enhanced whereas the other inhibited tumor growth. As immunoprophylactic agents, both phenotypes exerted, but at different degrees, antitumor protection in mice.     

Lipid Fatty Acid Composition and Thermophilicity of Cyanobacteria

An analysis of lipid fatty acid composition in several unicellular and filamentous forms of mesophilic and thermophilic cyanobacteria was performed. At 47°C (the temperature of thermophilic cyanobacteria maintenance in the collection), the unicellular thermophilic Synechococcus strains were devoid of polyenoic acids as distinct from the mesophilic forms of this genus at the temperature of 20°C (the temperature of this cyanobacterial maintenance in the collection). In the thermophilic Synechococcus elongatusIPPAS B-267 strain, a decrease in temperature did not result in the occurrence of C18 polyenoic acids, but the quantitative relationship between the saturated and unsaturated fatty acids (S/U ratio) was decreased twofold. In contrast, the culturing of mesophilic strains at 25–32°C resulted in an increase in the S/U ratio due to an increase in the proportion of the 16:0 acid. In the Synechococcus IPPAS B-434 strain, this treatment resulted in a decrease in the relative content of monoenoic, mainly hexadecenoic, acids. The cyanobacterium Gloeobacter violaceus, which lacks thylakoids, and whose photosystems are formed in a cell membrane, contained polyenoic acids. The filamentous thermophilic cyanobacterium Phormidium laminosum, at the maintenance temperature of 47°C, did contain polyenoic acids, but their proportion was considerably lower than that in the filamentous mesophilic forms, such as Tolypothrix sp. and Spirulina platensis. A relative content of hexadecenoic acids in Ph. laminosum was higher than in the mesophilic forms. A possible role of hexadecenoic acids in the processes of adaptation of cyanobacteria to high temperatures is discussed. A relationship between the characteristics of fatty acid composition fixed by evolution and the changes caused by adaptation to a particular environment is considered.     

Various biological properties of enterobacteria, isolated from patients with diarrhea

381 enterobacterial strains isolated from patients with acute enteric diseases were studied. Of these, 279 strains, as well as 20 strains isolated from 50 healthy children, were studied for the presence of adhesins, hemolysins and catalase. The comparison of the hemagglutinating activity of enterobacteria isolated from sick and healthy children revealed no essential differences between them. 15.8% of enterobacterial strains isolated from sick children possessed hemolytic activity, while strains isolated from healthy children did not induce the hemolysis of erythrocytes. All enterobacterial strains isolated from patients with acute enteric diseases were multi-resistant to antibiotics. A multitude of different antibiograms was obtained, most of them occurring only once. In 1985 the number of multiresistant (i. e. resistant to 11 and more antibiotics) strains dropped from 61% to 26..9% in comparison with 1981.