Synthesis, structure and properties of polyhydroxyalkanoates: biological polyesters

Recent advances in microbial polyhydroxyalkanoates

Poly-beta-hydroxybutyrate granules exhibited a strong orange fluorescence when stained with Nile blue A. Heat-fixed cells were treated with 1% Nile blue A for 10 min and were observed at an excitation wavelength of 460 nm. Glycogen and polyphosphate did not stain. Nile blue A appears to be a more specific stain for poly-beta-hydroxybutyrate than Sudan black B.

Nile blue A as a fluorescent stain for poly-beta-hydroxybutyrate.

The dormant and durable spore form of Bacillus anthracis is an ideal biological weapon of mass destruction. Once inhaled, spores are transported by alveolar macrophages to lymph nodes surrounding the lungs, where they germinate; subsequent vegetative expansion causes an overwhelming flood of bacteria and toxins into the blood, killing up to 99% of untreated victims. Natural and genetically engineered antibiotic-resistant bacilli amplify the threat of spores being used as weapons, and heighten the need for improved treatments and spore-detection methods after an intentional release. We exploited the inherent binding specificity and lytic action of bacteriophage enzymes called lysins for the rapid detection and killing of B. anthracis. Here we show that the PlyG lysin, isolated from the gamma phage of B. anthracis, specifically kills B. anthracis isolates and other members of the B. anthracis 'cluster' of bacilli in vitro and in vivo. Both vegetative cells and germinating spores are susceptible. The lytic specificity of PlyG was also exploited as part of a rapid method for the identification of B. anthracis. We conclude that PlyG is a tool for the treatment and detection of B. anthracis.

A bacteriolytic agent that detects and kills Bacillus anthracis.

Summary: A collection of 185 isolates of Pseudomonas solanacearum has been classified into 4 biotypes according to their capacity to oxidize 3 disaccharides (lactose, maltose and cellobiose) and 3 hexose alcohols (mannitol, sorbitol and dulcitol). Biotype 2 which oxidized the disaccharides but not the hexose alcohols appears to have a restricted host range; it was obtained solely from two plant hosts, potato and tomato, whereas biotype 1, which oxidized neither group of carbohydrates, and biotype 3 which oxidized both, were obtained from a diversity of plant hosts. A bacteriophage isolated from material infected with biotype 2 showed a high degree of specificity for isolates of biotype 2. The present known distribution of the 4 biotypes is given and their relationship to host range is discussed. A neotype culture for Ps. solanacearum is proposed.

Characteristics of Pseudomonas solanacearum

Following Hartman, who first suggested the use of Sudan black B, in place of red Sudans, as a bacterial fat stain (Hartman, 1940), Burdon, Stokes, and Kimbrough (1942a) confirmed the greater value of this dye and modified the procedure for demonstrating intracellular fatty material in bacteria by preparing, from suspensions of the organisms in alcoholic Sudan black B solution, dried films counterstained with safranine. Previously it had been thought that dried, fixed films were entirely unsuitable for fat stains (Lewis, 1941). These permanent films were regarded as an obvious improvement over the wet preparations used by earlier workers, and they were shown to be of practical aid in the classification of aerobic sporeforming bacilli (Burdon, Stokes, and Kimbrough, 1942b), but the staining method still had a number of undesirable features. Further experimentation has resulted in the development of the much superior procedure to be described here. The new method is not only simpler, requiring no more effort than a gram stain, but it is also far more rewarding, for the films now reveal clearly intracellular lipid matter that previously has not been seen or even suspected. The improved stain has increased differential value. More. over, its application, to various bacteria has resulted in certain general findings of unusual interest.

Fatty Material in Bacteria and Fungi Revealed by Staining Dried, Fixed Slide Preparations.

Experimental Infection of Mice with Bacillus cereus: Studies of Pathogenesis and Pathologic Changes

The methods suggested for the identification of species of the genus Bacillus in the invaluable monographs of Smith et al. (1946, 1952) have been applied to a collection of named cultures, and also have been utilized for the classification of approximately 400 strains of these organisms isolated from samples of blood, spinal fluid, surgical tissue, and other human and animal sources, as well as from laboratory dust, and soil. In the course of this work certain additional and modified techniques have been tested which appear to supplement those of Smith et al. in a useful way. In particular, the differential value of positive fat staining, of fermentation tests in tryptose agar, and of observations of the mode of spore germination has been appraised. Special procedures for differentiation of Bacillus licheniformis from Bacillus subtilis and Bacillus pumilus, and for the identification of both virulent and avirulent strains of Bacillus anthracis, have been tested. In the following report reference is made in detail only to the nine saprophytic species which in our experience are most likely to be encountered as contaminants from the laboratory dust, and in specimens from human beings or experimental animals, and which must be distinguished from B. anthracis. These species are B. cereus, B. mycoides (B. cereus var. mycoides, Smith et al., 1952), B. megaterium, B. subtilis, B. licheniformis, B. pumilus, B. circulans, B. brevis, and B. sphacericus.

Useful criteria for the identification of Bacillus anthracis and related species.

led Smith, Gordon and Clark (1952) to suggest that nonvirulent strains of B. anthracis may actually be identical with B. cereus, and further that it may be proper to regard virulent anthrax bacilli as pathogenic variants of the supposedly more stable parent species, B. cereus. Recently Brown et al (1955, 1956, 1958) have presented new evidence in support of these concepts. Brown, Moody, Treece and Smith (1958) described an anthrax-like disease produced in laboratory animals as the result of inoculating rapidly multiplying B. cereus which had apparently been transformed into capsulated bacilli capable of causing "the gross pathology of anthrax." In the present paper results of our further studies will be presented. By use of

On the differentiation of anthrax bacilli from Bacillus cereus.

https://www.jacionline.org/article/0021-8707(65)90192-9/pdf

Kenneth L. Burdon

Papers

Fatty Material in Bacteria and Fungi Revealed by Staining Dried, Fixed Slide Preparations.

Experimental Infection of Mice with Bacillus cereus: Studies of Pathogenesis and Pathologic Changes

Useful criteria for the identification of Bacillus anthracis and related species.

On the differentiation of anthrax bacilli from Bacillus cereus.

Observations on biochemical abnormalities in hereditary angioneurotic edema