Investigations of the pigments fromCytophaga johnsonae Cy j1

Besides carotenoids a complex of flexirubin-type pigments was isolated from the gliding bacteriumCytophaga johnsonae Cy j1 and separated into 6 components, which partly containe chlorine. In spite of the fact that these components still consist of pigment mixtures, the gross structures of 18 new flexirubin-type pigments could be deduced by spectroscopic and chemical investigations. The results open insights into biosynthesis and structural variety of the flexirubins, the novel non-isoprenoid pigments recently found inFlexibacter elegans.Non-Standard Abbreviations FT Fourier transformation - HPLC high pressure liquid chromatography - M⁺ molecular ion - M/M resolution of mass spectrometer - mu mass units - t _R retention time - TLC thin layer chromatography Part 16: Investigations on metabolites of microorganisms. Part. XV: H. Achenbach, W. Kohl, H. Reichenbach: Die Hauptpigmente vonCytophaga johnsonae. Tetrahedron Lett.1977, 1061. Part XIV: H. Achenbach, J. Witzke: Totalsynthese des Flexirubindimethylethers. Angew. Chem.89, 198 (1977); Angew. Chem. Int. Ed. Engl.16, 191 (1977)     

Light and Electron Microscopic Observations of Gametogenesis in Hastigerina pelagica (Foraminifera)*

SYNOPSIS. During gametogenesis mother individuals of Hastigerina pelagica (d'Orbigny) undergo significant morphological changes. Thirty h before gamete release, the cytoplasm changes from pale orange to bright red, possibly due to transport of stored lipids from the inner region to more peripheral parts of the cytoplasm. During the next 10 to 15 h the bubble capsule which surounds the calcareous shell is discarded. After all bubbles have disappeared, the individual sheds its spines by resorbing the spine bases close to the shell surface. A single mother nucleus divides into some hundreds of thousands of gamete nuclei within a span of ～ 20 h. A bulge of cytoplasm is extruded from the aperture and increases in size during the next 5 to 10 h. This bulge consists of cytoplasmic strands in which gametes and spherical bodies are embedded. The gametes and spherical bodies mature and are released during the afternoon and early evening. The gametes have 2 unequal acronematic flagella. A previously undescribed structure in foraminiferal reproduction is the spherical body which consists of a large vacuole surrounded by a thin cytoplasmic layer in which several nuclei, various typical cell organelles and multiple flagella are present. The spherical bodies are believed to play a role as receptacles of waste material, possibly including residual digestive enzymes, thereby protecting the gametes from lysis during the reproductive process. Fusion of gametes and further development into the next generation have not been observed.     

Decontamination of bacterial infection of monolayer cultures with a specific bacteriophage

Summary A few cell lines and primary monolayer cultures were accidentally infected by bacteria. These cultures were successfully decontaminated by means of the specific bacteriophage virus after quick identification of the responsible bacteria. This method presents a practical interest for preservation of valuable cultures. This work is supported by the Institut National de la Sante et de la Recherche Medicale (France) and the Fondation pour la Recherche Medicale (France).     

COLONY DEVELOPMENT IN EUDORINA ELEGANS (CHLOROPHYTA,VOLVOCALES)1

A detailed ultrastructure study was made of cell division and colony development in Eudorina elegans Ehrenberg. At the onset of cell division and prior to nuclear division the nucleus moved from the cell center to the cell surface. During nuclear division the nuclear membrane remained intact, except for openings occurring at the nuclear poles. The spindle microtubules appeared to arise from a MTOC-like (microtubule organizing centers) structure, while centrioles were absent from the nuclear poles. Following telophase, daughter nuclei formed which were separated by several distinct bands of endoplasmic reticulum. Cytokinesis occurred with formation of a cleavage furrow, associated with a typical phycoplast band of microtubules. However, cytokinesis was incomplete, resulting in formation of cytoplasmic bridges between the plakeal cells. Upon completion of up to five successive cell divisions, the plakea underwent inversion, which appeared to involve the production of colonial envelope material and rearrangement of cytoplasmic bridges. A new hypothesis concerning inversion is postulated based on these observations.     

Increased root exudation of14C-compounds by sorghum seedlings inoculated with nitrogen-fixing bacteria

Summary Organic components leaked fromSorghum bicolor seedlings (‘root exudates’) were examined by recovering¹⁴C labelled compounds from root solutions of seedlings inoculated withAzospirillum brasilense, Azotobacter vinelandii orKlebsiella pneumoniae nif-. Up to 3.5% of the total¹⁴C recovered from shoots, roots, and nutrient solutions was found in the root solutions. Inoculation with Azospirillum and Azotobacter increased the amounts of¹⁴C and decreased the amounts of carbohydrates in the root solutions. When sucrose was added as a carbon source for the bacteria, the increase of¹⁴C in the solutions did not occur. Quantities of¹⁴C found in the root solutions were proportional to amounts of mineral nitrogen supplied to the plants. Bacterial growth also was proportional to nitrogen levels. When sorghum plants were grown in soil and labelled with¹⁴CO₂, about 15% of the total¹⁴C recovered within 48 hours exposure was found in soil leachates.     

Studies on plating efficiency and estimation of viability of suspensions of Paracoccidioides brasiliensis yeast cells

Mild sonication was used to obtain single cell suspensions of Paracoccidioides brasiliensis. These cells were intact by microscopic criteria. Direct cell counts in a given inoculum and colony formation on various media were used to determine plating efficiency. Sonicated and nonsonicated cell suspensions were used to study plating efficiency and to estimate viability by means of vital dyes. Methylene blue, Erythrosin B, and Janus green were unreliable when used with P. brasiliensis, but vital dyes were accurate when tested with Candida albicans.Acridine orange gave more meaningful results of viability. Estimates of viability, however, changed significantly as a result of relatively minor alterations in the composition of the suspending medium.In initial experiments, the plating efficiency of P. brasiliensis was dismally low. It descended abruptly with increasing dilution of inoculum. Efficiency was much improved if horse serum was added to brain heart infusion plates or if glucose glycine yeast extract (GGY) plates were incubated at room temperature and mycelial colonies were counted. With the technique we report, current plating efficiency of sonicated suspensions is of the order of 25 %. Our results and procedures have an important bearing upon those studies concerned with in vitro killing of P. brasiliensis in suspensions or with isolating this fungus from clinical or environmental specimens.     

Ectothiorhodospira vacuolata sp. nov., a new phototrophic bacterium from soda lakes

A new phototrophic bacterium was isolated from Jordanian and Kenyan alkaline salt lakes. Cells are rod shaped, 1.5 m wide and 2–4 m long, and motile by polar flagella. They divide by binary fission, and possess photosynthetic membranes as lamellar stacks similar to those in the other species of the genus Ectothiorhodospira and the brown colored Rhodospirillum species. The presence of bacteriochlorophyll a and carotenoids of the normal spirilloxanthin series is indicated by the absorption spectra of living cells. Under certain growth conditions the cells form gas vacuoles, may become immotile and float to the top of the culture medium. Sulfide and thiosulfate are used as photosynthetic electron donors. During the oxidation of sulfide to sulfate, elemental sulfur is formed, which is accumulated outside the cells. The organisms are strictly anaerobic, do not require vitamins, are moderately halophilic and need alkaline pH-values for growth. The new species Ectothiorhodospira vacuolata is proposed.     

Autoinduction of bacterial bioluminescence in a carbon limited chemostat

Several strains of four species of luminous marine bacteria were maintained in a chemostat at a constant dilution rate and a variety of steady state densities by carbon (glycerol) limitation in order to study the relationship between culture density and bioluminescence activity. In general, luminescence per cell was constant at high culture density, and decreased dramatically at low culture density. For Vibrio fischeri, luminescence decreased to nondectable levels when the culture was maintained at low density; such dark cells were stimulated to synthesize luciferase and became luminous within minutes when purified autoinducer was added to the chemostat. Two strains, Photobacterium phosphoreum NZ11D and Photobacterium leiognathi S1, did not show the decrease in light intensity at low culture density that was characteristic of all other strains tested; they appeared to be constitutive for bioluminescence.Abbreviations BCM basal salts glycerol medium - BM basal salts medium - BSA bovine serum albumin - D dilution rate - DTT dilitiothreitol - LU light unit=2×10¹⁰ quanta s^-1 - OD optical density - SWC sea water complete medium - specific growth rate     

Acetogenium kivui,a new thermophilic hydrogen-oxidizing acetogenic bacterium

Hydrogen-oxidizing acetogenic bacteria in pure culture are presently represented by the two mesophilic species, Acetobacterium woodii and Clostridium aceticum. From Lake Kivu we have isolated a Gram negative, chemolithotrophic, thermophilic anaerobe (LKT-1) that oxidizes hydrogen and reduces carbon dioxide to acetic acid. It is a non-motile, non-sporeforming rod, about 0.7m in width and 2–7.5m in length, often occuring in pairs or chains. The cell wall has a banded appearance; the surface layer contains a regular array of particles with six-fold rotational symmetry. No outer membrane is present. The temperature optimum for growth is 66°C, and the pH optimum is 6.4. Organic growth substrates include glucose, mannose, fructose, pyruvate, and formate; acetate is the principal product. The doubling time for growth on hydrogen and carbon dioxide is about 2h. Vitamins are neither required nor stimulatory. Yeast extract and Trypticase enhance the final yield but do not affect the growth rate. Cysteine or sulfide are required and cannot be replaced by thioglycolate or dithiothreitol. LKT-1 was mass cultured on hydrogen and carbon dioxide in a 24.1 fermentor with a yield of 34g (wet weight) of cells. The DNA base composition as determined by buoyant density is 38 mol % guanine plus cytosine. LKT-1 appears only distantly related to physiologically similar bacteria. A new genus Acetogenium is proposed, and the species is Acetogenium kivui.     

耐药菌感染的噬菌体治疗专利技术

耐药菌,尤其是多重耐药菌的出现和持续进化给人类健康带来了巨大的威胁。在抗生素逐渐失去特效作用的情况下,科学界和医药界又把眼光重新投向了抗菌的天然生物-噬菌体,并在一些研究中证明了噬菌体可以作为新的武器去替代抗生素治疗耐药菌感染。通过对噬菌体治疗及衍生的裂解酶治疗的世界专利申请进行统计及分析,获得了专利发展趋势、申请人分布特点及主要专利申请人等信息,详细分析了噬菌体及裂解酶治疗的主要专利技术路线和研发热点。