Conjugal Transfer of Plasmid-Borne Multiple Antibiotic Resistance in Streptococcus faecalis var. zymogenes

A strain of Streptococcus faecalis var. zymogenes, designated JH1, had high-level resistance to the antibiotics streptomycin, kanamycin, neomycin, erythromycin, and tetracycline. These resistances were lost en bloc from approximately 0.1% of cells grown in nutrient broth at 45 C. The frequency of resistance loss was not increased by growth in the presence of the "curing" agents acriflavine or acridine orange, but after prolonged storage in nutrient agar 17% of cells became antibiotic sensitive. Covalently closed circular deoxyribonucleic acid (DNA) molecules were isolated from the parental strain and from antibiotic-sensitive segregants by using cesium chloride-ethidium bromide gradients. DNA molecular species were identified by using neutral sucrose gradients. Strain JH1 contained two covalently closed circular DNA species of molecular weights 50 x 10(6) and 38 x 10(6). An antibiotic-sensitive segregant, strain JH1-9, had lost the larger molecular species. A second sensitive segregant, strain JH1-5, had also lost the larger molecular species but a new molecular species of approximate molecular weight 6 x 10(6) was present. The antibiotic resistances that were curable from the parental strain were transferred to antibiotic-sensitive strains of S. faecalis and to strain JH1-9, during mixed incubation in nutrient broth at 37 C. Data to be described are interpreted to suggest that the transfer is by a conjugal mechanism. Analysis of the plasmid species in recipient clones showed that all had received the plasmid of molecular weight 50 x 10(6). Strain JH1-5 was not a good recipient. Analysis of one successful recipient clone of JH1-5 revealed that it had gained the 50 x 10(6) molecular weight plasmid but lost the 6 x 10(6) molecular weight species. These data are interpreted to mean that the multiple antibiotic resistance is borne by a transferable plasmid of 50 x 10(6) molecular weight, and that in clone JH1-5 this plasmid suffered a large deletion leaving only a 6 x 10(6) remnant which was incompatible with the complete replicon.     

Restricted pH ranges and reduced yields for bacterial growth under pressure

Hydrostatic pressure was found to cause a marked narrowing of pH ranges for growth and reductions in growth yields for a variety of bacteria. In many cases, reduced yields under pressure could be directly related to increased sensitivities to metabolic acids that accumulated in the enclosed culture vessels used. Magnesium and calcium ions partially reversed increases in sensitivities of representative gram-positive and gram-negative bacteria to low, but not high, pH. Growth inhibition of these organisms at both extremes of pH was associated with enhanced loss of K⁺ from pressurized cells. Inhibited cells in alkaline media also lysed under pressure, but microscopically observable lysis was clearly a secondary phenomenon because it occurred slowly. Apparent volumes for growth-inhibitory protonation-deprotonation reactions were calculated on the basis of measured shifts in inhibitory pH with pressure. The values ranged from 99 to 431 ml/mole, and their magnitudes indicated that growth inhibition by acids or bases involves cooperative changes in polymeric interactions such as those which accompany protein denaturation.     

Studies on the fractionation of total transfer ribonucleic acid and purification of an alanine transfer ribonucleic acid from chick embryo

Chick embryo tRNA, prepared by a simple large-scale method, was fractionated on three different ion-exchange columns. In all cases simple chromatographic patterns for various tRNA species were observed, indicating the presence of only a few major species of tRNA for each amino acid. By repeated chromatography one species of alanine tRNA was purified to approx. 80% purity. T₁ ribonuclease digest of this purified tRNA gave a simple chromatographic pattern. Because of the simplicity of the method of preparation of tRNA from this readily available source and the presence of only a few species of tRNA for each amino acid, chick embryo is suited for the study of tRNA and its various functions in higher systems.     

The effect of adrenocorticotrophin on protein degradation in rat adrenal gland and liver

The rate of adrenal protein degradation appears to be slower in rats to which ACTH (adrenocorticotrophin) has been chronically administered. As measured by the exponential decay of radioactively labelled adrenal protein in vivo, the mean half-lives of total protein and of mitochondrial, microsomal and 18000g-supernatant protein were significantly longer in ACTH-treated animals. Experiments in which either [(3)H]leucine or NaH(14)CO(3) was used to label proteins showed that of the fractions studied, the effect on mitochondrial protein degradation was most pronounced. The half-lives of the same subcellular fractions in rat liver were not affected by ACTH. The possibility that the results might have been caused by changes in radioisotope reutilization and pool size is discussed.     

Studies on the macronucleus of Paramecium aurelia

Following specific incorporation of tritiated thymidine into the DNA of the macronucleus of Paramecium aurelia for varying periods of time the vast majority of silver grains in electron microscope autoradiographs are associated with the area containing the small bodies and matrix. At best only 5% of the silver grains overlying the macronucleus are attributable to the large bodies. This is consistent with the view that the large bodies are the nucleoli of the macronucleus.     

Rumen Microbial Ecology in Mule Deer

Mule deer rumen microbial populations from animals in the natural habitat in Utah and from captive deer fed various rations were studied. The microorganisms were characterized on the basis of morphology and Gram reaction. Rumen samples contained 13 identifiable types of bacteria and one genus of ciliate protozoa (Entodinium). Highest rumen bacterial populations were produced on rations containing barley. No differences in proportions of ruminal bacteria in the various morphological groups could be detected when animals were fed either natural browse plants or alfalfa hay. The total numbers of bacteria were similar for animals feeding on controlled diets of browse or hay and those in the natural habitat. Numbers of some bacterial types were directly related to ciliate protozoal numbers, whereas others were inversely related. Highest rumen ciliate protozoal populations were observed on rations containing barley. No differences in protozoal populations were noted between diets containing only browse or hay. Seasonal variations were noted in ciliate protozoal numbers from deer feeding in the natural habitat. The total number of ciliate protozoa decreased in the fall and winter and remained low until spring. There were indications that salt in the deer diet favorably affected rumen ciliate protozoa. Rather than revealing direct deer management applications, this study serves to stimulate and illuminate new approaches to research in range and wildlife nutrition.     

Zur Feinstruktur von Cuticula und Epidermis beim Flußkrebs Orconectes limosus während eines Häutungszyklus

Zusammenfassung Die Cuticula an der Innen- und Außenseite der Branchiostegite des Flußkrebses besteht wie für Arthropoden üblich aus Epi- und Procuticula. Sowohl die Epicuticula als auch die Procuticula von Innen- und Außenseite unterscheiden sich im Feinbau wesentlich voneinander. An der Innenseite ist die Epicuticula einfach gebaut; Die Procuticula ist lamelliert und zeigt meist die bogenförmigen Muster von Mikrofibrillen. Die Epicuticula an der Außenseite weist in den in dieser Arbeit untersuchten Entwicklungsstadien einen sehr viel komplizierteren Feinbau auf, der in der Entwicklung gewissen Änderungen unterliegt. In der wiederum lamellierten Procuticula an der Außenseite sind die Mikrofibrillen zu Balken gebündelt. Die Ausrichtung der Mikrofibrillen dreht sich innerhalb einer Lamelle um 180°. Durch die Procuticula ziehen Fortsätze der Epidermiszellen, außerdem Stäbe der sog. Verbindungsstrukturen.Die Bildung der Cuticula an der Innenseite konnte weitgehend vollständig verfolgt werden; sie ist gut mit der Bildung der Cuticula bei verschiedenen Insekten vergleichbar.Die Bildung der Cuticula an der Außenseite konnte dagegen nur von Beginn der Abscheidung der Proouticula bis zur Häutung verfolgt werden. Kurz vor Beginn der Cuticulaabscheidung kommt es in den Epidermiszellen zu einer stärkeren Entwicklung des rauhen ER. Während der gesamten von uns verfolgten Bildungsstadien sieht man Vesikel mit dichtem Inhalt besonders in der Nähe des Zellapex. Sie geben anscheinend hier ihren Inhalt, Cuticulamaterial, nach außen ab. Sie stammen wohl aus Golgibereichen. Auch Stachelsaumbläschen (coated vesicles) kommen regelmäßig vor, deren genetischer Zusammenhang mit multivesikulären Körpern diskutiert wird. Bei der Abscheidung der fibrillären Cuticulasubstanzen spielen besondere Differenzierungen der Zell oberfläche, — kappenartige Verdichtungen der Zelloberfläche, meist an der Spitze kleiner Mikrovilli — eine wesentliche Rolle.

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The ultrastructure of cuticle and epidermis in the crayfish Crconectes limosus during a moulting cycle

Summary The cuticle of the inside and outside of the branchiostegites of the crayfish consists of an epicuticle and a procuticle — as common in arthropods. Concerning their ultrastructure epicuticle and procuticle differ essentially from each other on both the inside as well as the outside. On the inside the epicuticle is built plainly; the procuticle is laminated, and, mostly it shows the arched patterns of microfibrils. In those developmental stages investigated in this project the epicuticle of the outside shows a much more intricated ultrastructure, since during formation it is subject to certain changes. On the outside the procuticle is also laminated; the microfibrils are bundled up to bars. The alignment of those microfibrils within one lamella is twisted for 180°. The procuticle is penetrated by processes of epidermal cells and by rods of the so-called connecting structures.The formation of the cuticle on the inside was observed completely; it is comparable to the forming of the cuticle in several insects. However, the formation of the cuticle on the outside was only observed from the beginning of the procuticular development up to the moulting.Shortly before formation of the cuticle the development of rough ER in the epidermal cells seems to be intensified. In all of developmental stages observed there appear vesicles with dense contents mainly situated nearby the cell apex. At this site they evidently deliver their contents — cuticular materials — to the outside of the cell; they probably originate in the Golgi areas. There occur coated vesicles regularity, too; their genetic relation to multivesicular bodies is discussed. Special differentiation on the cell surface i.e. dome-like consolidations of the cell surface mainly placed at the tip of small microvilli are of great importance for the secretion of the cuticle substances.