Somatic antigens of Pseudomonas aeruginosa. The structure of the O-specific polysaccharide chains of P. aeruginosa O11 (Lányi) lipopolysaccharides

Lipopolysaccharides were isolated from the phenol layer on aqueous phenol extraction of cells of Pseudomonas aeruginosa O11 (Lányi classification), strains 170021 and 170040. On mild acid degradation of the lipopolysaccharides, with the subsequent gel-filtration on Sephadex G-50, neutral O-specific polysaccharides made up of 6-deoxysugars alone were obtained. Two 2-acetamido-2,6-dideoxy-L-galactose (LFucNAc), 2-acetamido-2,6-dideoxy-D-glucose (DQuiNAc) and L-rhamnose (LRha) residues were found to be the components of the strain 170021 polysaccharide repeating units; those of strain 170040 contained the same monosaccharides, but, instead of 2-acetamido-2,6-dideoxy-D-glucose residue, that of 2-acetamido-2,6-dideoxy-D-galactose (DFucNAc) was present. On the basis of the 13C nuclear magnetic resonance data, methylation analysis and three successive Smith degradations the following structures were determined for the polysaccharide repeating units: strain 170021----2) LRha(alpha 1----3)LFucNAc(alpha 1----3)LFucNAc(alpha 1----3)DQuiNAc(beta 1----; strain 170040,----2)LRha(alpha 1----3)LFucNAc-(alpha 1----3)LFucNAc(alpha 1----3)DFucNAc(beta 1----; differing from one another by configuration of C-4 of 2-acetamido-2,6-dideoxy-D-hexopyranose only.     

Somatic antigens of Pseudomonas aeruginosa. The structure of the O-specific polysaccharide chains of lipopolysaccharides of P. aeruginosa serogroup O4 (Lányi) and related serotype O6 (Habs) and immunotype 1 (Fisher)

Acidic O-specific polysaccharides were isolated on mild acidic degradation of lipopolysaccharides of Pseudomonas aeruginosa serotypes O4a,b, O4a,c, O4a,d (Lányi classification) and serologically related to them serotype O6 (Habs classification) and immunotype 1 (Fisher classification). The polysaccharides had identical monosaccharide composition and were built up of L-rhamnose, 2-acetamido-2,6-dideoxy-D-glucose,2-formamido-2-deoxy-D-galacturonic acid and 2-acetamido-2-deoxy-D-galactouronamide residues. The latter two derivatives of D-galactosaminuronic acid were found in nature for the first time. All the polysaccharides, but Lányi serotype O4a,c, contained O-acetyl groups. The polysaccharides were readily de-O-acetylated with aqueous triethylamine and de-N-formylated with dilute hydrochloric acid. De-N-formylated polysaccharide of serotype O4a,c was selectively cleaved with nitrous acid upon 2-amino-2-deoxygalacturonic acid residues to form a tetrasaccharide with a 2,5-anhydrotaluronic acid residue on the reducing end. The tetrasaccharide represented a modified repeating unit of the polysaccharide. Solvolysis of all intact polysaccharides with hydrogen fluoride selectively split the glycosidic linkages of 6-deoxy sugars to give the same trisaccharide, including both derivatives of galactosaminuronic acid and having 2-acetamido-2,6-dideoxyglucose on the reducing end. Structural investigation of the oligosaccharides obtained together with methylation analysis and 13C nuclear magnetic resonance data revealed the following structures of the O-specific polysaccharides: (Formula: see text) An independent confirmation of the structures of the repeating units was obtained as the result of full interpretation of the 13C nuclear magnetic resonance spectra of the intact and modified polymers. Spectral data analysis revealed a number of regularities in the effects of glycosidation connecting their values with the anomeric and absolute configuration of pyranose residues. The data on the structures of the O-specific polysaccharides indicated that each of the five P. aeruginosa strains under study should be considered as an individual O-serotype within one O-serogroup.     

Antigenic polysaccharides of bacteria. 18. Structure of O-specific polysaccharide chains of Pseudomonas aeruginosa 05 (Lányi) lipopolysaccharide

Polysaccharide chains of P. aeruginosa O5a, b, c, O5a, b, d and O5a, d (Lányi classification) lipopolysaccharides contain D-xylose, N-acetyl-D-fucosamine (FucNAc) and a derivative of 5,7-diamino-3,5,7,9-tetradeoxy-L-glycero-L-manno-nonulosonic acid (pseudaminic acid, PseN2) carrying acetyl or (R)-3-hydroxybutyryl (Hb) and formyl (Fm) groups as N-acyl substituents. Degradation of the lipopolysaccharides with dilute acetic acid caused depolymerisation of the polysaccharide chains as a result of cleavage of glycosidic linkage of pseudaminic acid to give trisaccharides representing chemical repeating units of the polysaccharides. Basing on analysis of the trisaccharides using 1H and 13C NMR spectroscopy and mass-spectrometry, the following structures of the polysaccharide chains were established: (Formula: see text). O5a, d polysaccharide is identical to P. aeruginosa immunotype 6 O-specific polysaccharide.     

Subunit structure and higher order assembly of the hemocyanins of the Melongenidae family: Melongena corona (Gmelin), Busycon canaliculatum (Linné), B. carica (Gmelin), B. contrarium (Conrad), and B. spiratum (Lamarck)

1. The hemocyanins of the Melongenidae family of marine gastropods: Melongena corona, Busycon canaliculatum, B. carica, B. contrarium, and B. spiratum exist in solution as multi-decameric aggregates characterized by sedimentation coefficients of approximately 105 S, 130 S, 150 S, 170 S, and higher values, corresponding to di-, tri-, tetra-, penta-, and larger multi-decameric particles. 2. The hemocyanins of B. contrarium and B. carica seem to form the largest decameric aggregates with the tri- to penta-decamers respresenting the major constitutents. Scanning transmission electron microscopy (STEM), both of unstained, freeze-dried and negatively-stained specimens, shows the presence of discrete aggregates consisting of up to ten decameric units. 3. The particle masses as determined by STEM mass measurements for individual molecules gave integral multiples of from 4.2 x 10(6) to 4.4 x 10(6) daltons ranging from about 8.2 x 10(6) daltons for the typical di-decamer of B. canaliculatum hemocyanin to as high as about 39 x 10(6) and 43 x 10(6) for the nano-and deca-decamers of B. contrarium hemocyanin. 4. The appearance of the higher multi-decamers in both negatively-stained and freeze-dried specimens suggest that they are formed by the addition of decameric units to a single di-decameric unit "tail-wise" in both directions. The higher aggregates formed seem to terminate with a closed head or collar at both ends of the assembly.     

The morphology and fine structure of the colourless colonial flagellate Anthophysa vegetans (O. F. Müller) Stein

A strain of Anthophysa vegetans (O. F. Müller) Stein (Chrysophyceae or Chrysomonadida) was grown in culture and examined in detail by light and electron microscopy. It was shown to be a typical, though apochlorotic, member of its class; features of interest include a well-developed leucoplast and a starch-like reserve product. Aspects of the largely inorganic stalk are also discussed.     

Lucilia sericata strain from Colombia: Experimental colonization, life tables and evaluation of two artificial diets of the blowfly Lucilia sericata (Meigen) (Diptera: Calliphoridae), Bogotá, Colombia strain

The objective of this work was to establish, under experimental laboratory conditions, a colony of Lucilia sericata, Bogotá-Colombia strain, to build life tables and evaluate two artificial diets. This blowfly is frequently used in postmortem interval studies and in injury treatment. The parental adult insects collected in Bogotá were maintained in cages at 22°C±1 average temperature, 60%±5 relative humidity and 12 h photoperiodicity. The blowflies were fed on two artificial diets that were evaluated over seven continuous generations. Reproductive and population parameters were assessed. The life cycle of the species was expressed in the number of days of the different stages: egg = 0.8±0.1, larvae I = 1.1±0.02, larvae II = 1.94±0.16, larvae III = 3.5±0.54, pupae = 6.55±0.47, male adult = 28.7±0.83 and female adult = 33.5±1.0. Total survival from egg stage to adult stage was 91.2% for diet 1, while for diet 2 this parameter was 40.5%. The lifetime reproductive output was 184.51±11.2 eggs per female. The population parameters, as well as the reproductive output of the blowflies that were assessed, showed relatively high values, giving evidence of the continuous increase of the strain over the different generations and making possible its maintenance as a stable colony that has lasted for more than two years.     

Selective advantage of ray florets in Scalesia affinis and S. pedunculata (Asteraceae), two endemic species from the Galápagos

The presence of neuter ray florets in species within Asteraceae is generally believed to increase pollinator attraction. In the endemic Galápagos genus Scalesia (Asteraceae) a natural variation in the presence/absence of neuter ray florets is found. To evaluate whether the presence of ray florets plays a selective role on female reproductive success we chose two species of Scalesia, Scalesia affinis that carries ray florets and S. pedunculata that is rayless. On Santa Cruz Island capitula of S. pedunculata were equipped with fake ray florets while others were untouched. On Isabela Island ray florets were removed on half of the capitula of S. affinis. In S. affinis rayed capitula received more pollinators and more pollen, which resulted in a significantly higher embryo production. In S. pedunculata no effect on embryo production was found. The disagreement between the two species may correspond to a difference in visitation frequency, S. pedunculata receiving many more visit than S. affinis. Thus, ray floret development proved beneficial in pollinator-restricted localities.     

On the plurality of Chydorus sphaericus (O. F. Müller) (Cladocera,Chydoridae), and designation of a neotype from Sjaelsø, Denmark

Two readily distinguishable species of Chydorus sphaericus sens. lat. occur in Salmon Lake, Montana, differing from each other in size and shape of rostrum, headpore configuration, frequency of doubling of teeth on the postabdomen, pattern of reticulation of the shell, and morphology of the mature male, especially the postabdomen, postabdominal claws, copulatory hooks, and rostrum. In any such comparisons, individual specimens are not sufficient; populations are needed to sort out morphological characters associated with instar and sex and character's that change with increasing body size. Comparison of population structure and stage of reproduction can be accomplished by a size-frequency distribution and by specific morphological characters that enable the three male instars to be recognized individually.One species, thought at first to be Chydorus herrmanni, is very close to Chydorus sphaericus sens. str. from Denmark. The other taxon with a short, generally blunt labrum, is described as the new species Chydorus brevilabris. Because of morphological diversity among the entities currently listed as Chydorus sphaericus from around the World, it is certain that a complex of species is represented. To facilitate the eventual resolution of this problem, the population in Sjaelsø, Sjaelland, Denmark has arbitrarily been selected as the standard of comparison, and from this population a neotype and several allotypes have been designated. Cursory examination of various available populations suggests that C. sphaericus sens. str. and C. brevilabris represent a major dichotomy in the sphaericus complex, each branch consisting of an unknown number of closely-related species. Cautions are voiced against assuming that Chydorus sphaericus is a single cosmopolitan and ubiquitous species.This study has been aided by grant GB-29725 for the work in Denmark in 1971–72 and by my present grant DEB 76-20237, both from the National Science Foundation. I am indebted to D. F. Brakke for the samples from Salmon Lake and neighboring localities in Montana and information concerning these localities, and to Brenda J. Hann for dissections and drawings of trunk limbs and help in their interpretation. I am also indebted to Geoffrey Fryer, whose innocent inquiry precipitated this study.     

Isolation and identification of the antialgal compound, harmane (1-methyl-β-carboline), produced by the algicidal bacterium, Pseudomonas sp. K44-1

Twelve strains of algicidal bacteria were isolated from the surfacewater of the pond Shinobazu and the moat Ote-bori (Tokyo, Japan). Nine of thesestrains were considered to be in the Pseudomonas group byanalyses of 16S rDNA sequences. The methanol extract ofPseudomonas sp. K44-1 showed marked antialgal activity bythe paper disk method. Harmane (1-methyl--carboline) was isolated fromtheethyl acetate extract of the whole culture broth ofPseudomonas sp. K44-1 by using silica gel columnchromatography and high performance liquid chromatography (HPLC). Harmaneshowedantagonistic activities against several cyanobacterial strains at aconcentration of 30 g disk^–1.     

Depth-dependent reproductive output of the green sea urchin, Strongylocentrotus droebachiensis (O.F. Müller), in relation to the nature and availability of food

Reproductive output of green sea urchins (Strongylocentrotus droebachiensis O.F. Müller) in the field was highest at depths where preferred macro-algae were abundant, and lowest at depths where preferred macro-algae were overgrazed or replaced by non-preferred species (Agarum and Ptilota). Feeding rate and gonad indices of sea urchins in the laboratory were highest on a diet of preferred algae (Fucusdistichus L. subsp. edentatus (Pyl.) Powell, Laminaria longicruris Pyl., Desmarestia spp. and Saccorhizadermatodea (Pyl.) J. Ag.), and lowest on the less preferred Agarum cribrosum (Mert.) Bory, Ptilota serrata Kütz., and crustose corallines. Gamete production/unit area in overgrazed habitats was as great or greater than in kelp beds because of the higher biomass of urchins in overgrazed areas. Gonad weight and reproductive output of urchins from habitats poor in food can be increased by providing preferred foods.     

Habitat use, size structure and sex ratio of the spot-legged turtle, Rhinoclemmys punctularia punctularia (Testudines: Geoemydidae), in Algodoal-Maiandeua Island, Pará, Brazil

Rhinoclemmys punctularia punctularia is a semi-aquatic chelonian found in Northern South America. We analyzed the habitat use, size structure and sex ratio of the species on Algodoal-Maiandeua Island, a protected area on the Northeastern coast of the Brazilian state of Pará. Four distinct habitats (coastal plain lake, flooded forest "igapó", interdunal lakes, and tidal channels) were surveyed during the rainy (March and April) and dry (August and September) seasons of 2009, using hoop traps. For the analysis of population structure, additional data were taken in March and August, 2008. A total of 169 individuals were captured in flooded forest (igapó), lakes of the coastal plain and, occasionally, in temporary pools. Capture rates were highest in the coastal plain lake, possibly due to the greater availability of the fruits that form part of the diet of R. p. punctularia. Of the physical-chemical variables measured, salinity appeared to be the only factor to have a significant negative effect on capture rates. The sex ratio was only slightly biased to females, and did not vary between habitats or seasons. Straight carapace length was significantly larger in females, but did not vary between habitats. Overall, the evidence indicates that both biotic and abiotic factors like food availability, low current and salinity, influence the habitats selection and use by R. p. punctularia on Algodoal-Maiandeua Island.     

Fine structure of the eggshells of four primitive moths: Hepialus hecta (L.),Wiseana umbraculata (Guénée) (Hepialidae), Mnesarchaea fusilella walker and M. Acuta philp. (Mnesarchaeidae) (Lepidoptera,Exoporia)

The eggshells of Hepialus hecta, Wiseana umbraculata (Hepialidae) and of Mnesarchaea fusilella (Mnesarchaeidae) (Lepidoptera, Exoporia) were studied by scanning electron microscopy (SEM) and transmission (TEM) electron microscopy. All 3 species show a very similar surface sculpture of the micropylar region which, however, is very different from the eggs of the Ditrysia. The micropylar plate is large and oval. There are only 2 or 3 micropylar openings. In the hepialid moths, the surface of the egg's main body is characterized by spherical protuberances. The radial fine structure of the eggshells of all 3 species as well as of alcohol-stored eggs of Mnesarchaea acuta (Mnesarchaeidae) is basically identical to the eggshell of Korscheltellus lupulinus (Hepialidae). The chorion consists of only one layer, which, in the hepialid species, shows a crystalline-like fine structure. The vitelline envelope is composed of a thin, laminated, outer layer (V-2) and a thick, rigid, inner layer (V-1) that is traversed by large numbers of canals. This kind of eggshell architecture is distinctively different from that of the ditrysian Lepidoptera.