Staphylococcus aureus at a maternity ward. II. Characterization of isolates by enterotoxigenicity and toxic shock syndrome toxin (TSST-1) production

Tests for enterotoxins A, B, C, D, E and TSST-1 production were carried out on 775 S. aureus strains isolated from various sources (50 mothers and neonates studied periodically, mothers and infants treated for various acute inflammatory conditions, members of hospital staff, environmental swabs) during the period 1981-1983 at a maternity ward chosen for a 3-year systematic study and on additional 97 isolates obtained in 1985 from another maternity ward. This had contributed to a better classification of strains within certain phage type groups. It was found that the distribution of S. aureus types in the particular sub-sets varied, depending on the source of isolates. At the maternity ward followed for 3 years there was a clear-cut trend towards the spread of phage-untypable isolates producing enterotoxin C whereas at ward examined for comparative purposes B enterotoxin producers of phage type 95 were predominant. The tests for enterotoxigenicity has also proved to be useful as the epidemiological marker characterizing the predominantly circulating S. aureus strain. It has been confirmed that the majority role in the spread of maternity-ward-staphylococci is played by the neonates and the factors of hospital environment.     

Microflora participating in the decomposition of carboxymethyl cellulose continuously added to the soil

The microbial community in the soil was analyzed during four weeks of a continuous enrichment of structural chernozem soil samples with a 0.1% solution of carboxymethyl cellulose (CMC) under aerobic and semianaerobic conditions. During the first 14 d, the total amount of the aerobic and anaerobic, cellulose-degrading microorganisms increased significantly. Various metabolic pathways were u‘ed te decompose the substrate: diverse metabolic systems were activated and different groups of microorganisms preferred in dependence on the presence of oxygen or the source of mineral nitrogen. In the later phases of cultivation, a decrease in the concentration of zymogenous microflora and in the level of substrate mineralization was observed ovon though CM-cellulase activity remained high. During the fourth week of cultivation, a conspicuous increase in the numbers of oligothropic bacteria occurring in the colcnies of the microorganisms degrading cellulose was found. The representatives of prosthecobacteria (Caulobacter, Hyphomicrobium, Prosthecomicrobium spp.) andSeliberia sp. were thus identified. This “microflora of dispersion” attends the zymogenous microbes degrading CMC and indicates later phases of the process of decomposition.     

Poly-3-hydroxybutyratc production and changes of bacterial community in the soil

Changes in the number of bacteria and relative distribution of strains producing poly-3-hydroxybutyrate (PHB) in soil were investigated. Samples of chernozem soil were cul tivated with glucose in the presence of a mineral nitrogen source (diammonium hydrogen phos phate) or in its absence, either in a batch or a heterocontinuous cultivation system. In both cultivation systems the addition of glucose resulted in a roughly ten-fold increase of bacteria concentration and an increase in the proportion of strains able to produce PHB granules. When the nitrogen source was added simultaneously with glucose, the concentration of bacteria increased by two orders of magnitude in both cultivation systems. In the batch system changes in the concentration of strains capable of PHB production were very small under these con ditions whereas in the heterocontinuous system their number decreased by almost 50 %. The survival of bacteria in the soil suspension after 57-d starvation was associated with PHB production which differed, depending on the previous treatment of the soil samples. The concentration of bacteria decreased least pronouncedly in the control with water and most significantly during cultivation with glucose and a nitrogen source, where the initial PHB content was very low in spite of high numbers of bacteria.     

Effect of glucose on the amount of bacteria mineralizing 2,4-dichlorophenoxyacetic acid in soil

Plate numbers of bacteria and relative incidence of strains capable of mineralization of 2,4-dichlorophenoxyacetic acid (2,4-D) in chernozem samples incubated for 14 d with the herbicide (50 ppm) in the presence or absence of glucose (1000 ppm) were compared. Whereas the total number of bacteria increased 1.2-fold in the variant with 2,4-D and 2.4-fold in the variant with glucose and the herbicide, the number of 2,4-D-mineralizing bacteria increased 12.1-fold and 34.2-fold, respectively. In a collection of 96 isolates of soil bacteria substantially more strains capable of degradation of 2,4-D in the presence of glucose were detected as compared with the variant without it, indicating that processes of cometabolic type are involved during the degradation of this herbicide in the soil.     

Microbial decomposition of coumarin in soil

Microbial decomposition of coumarin was studied in samples of chernozem soil by manometric measurement of oxygen consumption, paper chromatography of aromatic metabolic intermediates in soil extract and measurement of their UV spectra, and by the technique of simultaneous adaptation. Coumarin is decomposed in soil viao-coumaric and melilotic acids and at least one other compound of aromatic character. The metabolic pathway including salicylic acid and catechol was not proved. A total of 39 strains of coumarin-decomposing bacteria were isolated from the soil, out of which 25 belong to the genusPseudomonas, 7 to the genusCellulomonas and 7 to the genusAchromobacter. A comparison of the counts of bacteria utilizing coumarin as a sole carbon source in garden soil, in two chernozem soil samples and in acidic brown soil showed that their occurrence bears no relation to the so-called total number of bacteria (grown on agar medium with yeast and soil extracts and with tryptone) or to the content of carbon and nitrogen in the soil, or to its acidity.     

Presence and oxidation of amino acids in rhizosphere and non-rhizosphere soil

Manometric studies were carried out on the respiratory activity of different rhizosphere and non-rhizosphere soils to follow quantitatively the over-all microbial activity in the rhizosphere soil as affected by the species and growth phase of plant. Oxygen consumption was distinctly greater in rhizosphere soils as compared to that in non-rhizosphere soils. The difference between oxygen consumption by rhizosphere and non-rhizosphere soils changed with the course of plant growth and it was not the same in different plants. This difference in oxygen consumption might be a measure of the amount of available oxidizable substrate in the rhizosphere. A rhizosphere sample had greater diversity as well as higher concentration of free amino acids than a non-rhizosphere sample of the same soil. Bacterial counts pointed to preferential stimulation in the rhizosphere of bacteria requiring individual amino acids. amino acids, such as glycine, alanine, asparsic acid of tyrosine were oxidized more rapidly in rhizosphere than in non-rhizosphere soil, but the extsent of oxidation for each of the amino acids studied did not differ. The amount of oxygen consumed during oxidation of alanine, aspartic acid or tyrosine was about one-half of the total amount necessary for complete oxidation. With glycine a higher extent of oxidation (60%) was observed. These extents of oxidation of glycine and aspartic acid did not change on investigation at two different phases of plant growth.     

A propos des ≪chloride cells≫ dans l'épithélium des lamelles branchiales du poisson rouge

Résumé L'ultrastructure des lamelles branchiales et spécialement celle des chloride cells du poisson rouge (Carassius aureus) a été étudiée. Nous avons constaté que du matériel amorphe floconneux, faiblement adiélectronique était attaché aux endroits des creux apicaux. Afin de préciser la nature de ce matériel, nous avons étudié ces structures au microscope électronique avec les techniques suivantes: acide periodique méthènamine d'argent, colorations au fer colloïdal et au bleu d'alcian. Après la réaction à l'acide periodique méthènamine d'argent, de fines précipitations aux endroits des creux apicaux, correspondant au matériel floconneux visible après la fixation au glutaraldéhyde tétroxyde d'osmium, étaient visibles. La coloration au bleu d'alcian révélait des particules fortement colorées formant un film plus ou moins continu à la surface libre des lamelles, sauf aux endroits oò les chloride cells sont en contact avec la surface. Là et notamment dans les 2reux apicaux, du matériel légèrement granuleux, de faible densité, faisait une couche assez épaisse attachée à la membrane cellulaire. Tenant compte des résultats d'autres auteurs et de nos propres observations, nous considérons que la plus grande partie du matériel se trouvant à la surface des chloride cells, et particulièrement dans les creux apicaux, est de type glycoprotéique.

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The ultrastructure of the chloride cells in the gill epithelium of the goldfish

Summary The ultrastructure of the secondary lamellae of the gills and especially that of the chloride cells of Carassius aureus was studied. We found an amorphous, flakey, slightly adielectronic material in the areas of the apical pits. In order to determine the nature of this material, we studied these structures electronmicroscopically applying the periodic acid silver methenamine, colloidal iron and alcian blue methods. The periodic acid silver methenamine reaction, resulted in finely dispersed precipitations which were deposited in the areas of the apical pits and which correspond to the flakey material seen in the ordinary electron micrographs. The alcian blue method reveales strongly stained particles which form a more or less continuous film on the free surface of the lamellae, interrupted only at the level of the chloride cells. In these areas, notably within the apical pits, a rather thick layer of finely granular low-density material is attached to the plasma membrane. In taking into account other studies performed on this subject, as well as our own observations, we consider the material found on the surface of the chloride cells and particularly within their apical pits to be predominantly of glycoproteinous nature.

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Dédié à Monsieur le Professeur Dr Ernst Horstmann, Hambourg, à l'occasion de son soixantième anniversaire.