MECHANISM OF ACTION OF THE TOXIN OF BACILLUS ANTHRACIS II. : Alkaline Phosphatasemia Produced by Culture Filtrates of Various Bacilli.

Slein, Milton W. (Fort Detrick, Frederick, Md.) and Gerald F. Logan, Jr. Mechanism of action of the toxin of Bacillus anthracis. II. Alkaline phosphatasemia produced by culture filtrates of various bacilli. J. Bacteriol. 83:359-369. 1962.-A factor which produces hyperphosphatasemia after intravenous injection into animals has been found in culture filtrates of several bacilli. The factor appears not to be lecithinase, although it has been found only in culture filtrates of microorganisms, such as Bacillus cereus variants and B. thuringiensis, which are known to produce lecithinase. The factor has been shown to be different from the protective antigen fraction of the toxin of B. anthracis. The phosphatasemic response can be prevented by mixing the factor with antiserum before injection.Immunological tests indicate that the centers of ossification may be a principal source of the excess alkaline phosphatase which is liberated into the bloodstream. However, moderate changes in bone alkaline phosphatase activity levels, brought about by the development of scurvy in guinea pigs or by treatment of scorbutic guinea pigs with ascorbic acid, did not seem to affect the phosphatasemic response of the animals. Marked release of alkaline phosphatase occurs when slices of rabbit epiphyseal bone or kidney cortex, or suspensions of leukocytes, are incubated with small amounts of the phosphatasemia factor.     

Continuous Culture of Ruminal Microorganisms in Chemically Defined Medium: II. Culture Medium Studies

Ruminal ciliates have been grown in continuous culture in chemically defined media and in the absence of viable bacteria. Oligotrichic ruminal ciliates seem to require insoluble carbohydrates for growth; the holotrichic ciliates require soluble carbohydrates, but at low concentrations. Both groups of ciliates utilize amino acids as their principal nitrogen source when these are supplied in micromolar concentrations; at millimolar concentrations, amino acids are toxic, possibly from excessive ammonia formation arising from ciliate deaminase activity. Holotrichic ruminal ciliates are destroyed by overdeposition of amylopectin when glucose is present above 0.1% concentration in the medium. Ecological requirements of ruminal ciliates are also described.     

Pyrogenic Responses to Staphylococcal Enterotoxins A and B in Cats

Pyrogenic responses, ranging up to 4.8 F, were induced in cats by oral administration of highly purified staphylococcal enterotoxin B in doses from 10 to 100 mug/kg. Fever was a more sensitive indicator of intoxication than was emesis. Highly purified preparations of enterotoxin A, whether administered intravenously (0.01 to 1.0 mug/kg), orally (10 to 25 mug/kg), or into the cerebral ventricles (0.005 to 0.020 mug in 0.20 ml), were also pyrogenic in cats. Tolerance to the pyrogenic activity was produced by repeated intravenous injection of a given dose of enterotoxin A but not by repeated intracerebroventricular injection. Enterotoxin A was more potent than enterotoxin B after intravenous injection in causing both fever and emesis. Cross-tolerance could not be demonstrated between enterotoxin A and enterotoxin B or Salmonella typhosa endotoxin. This lack of cross-tolerance plus the inability of large oral doses (100 to 4,700 mug/kg) of endotoxin to cause fever or emesis indicate that the reported responses were attributable to the specific toxins administered and not to contamination by other pyrogens.     

Cellulolytic Enzyme System in Culture Filtrates of Aspergillus sydowi

During growth in liquid culture medium, that contained single soluble or insoluble cellulosic carbon source, Aspergillus sydowi (Bain. & Start.) Thom & Church released cellulolytic enzymes into the medium. The enzymes were separated by gel filtration followed by ion exchange chromatography into three components, all of high molecular weight. One of the components (Ac) has the character of a C₁ cellulase enzyme. In the assay for hydrolysis of insoluble cellulose, the combined fractions, especially whenever the fraction under test contained the component Ac, released more glucose than when each component was employed alone.     

In Vivo Studies on Staphylococcal Penicillinase.

Eyckmans, Luc (Baylor University Medical Center, Dallas, Tex.), and Ralph Tompsett. In vivo studies on staphylococcal penicillinase. J. Bacteriol. 90:589-593. 1965.-The amounts of staphylococcal penicillinase present in tissue of mice with well-established infections due to penicillin-resistant staphylococci were determined. The data indicate that, in the presence of such infection, penicillinase can be found in amounts adequate to destroy large amounts of penicillin. These results are found in the absence of any conditions which are known to induce penicillinase. Although it has been generally assumed that penicillinase does account for the penicillin resistance of certain staphylococci in vivo, the data presented here document the quantitative aspects of this phenomenon, and indicate that the amounts of penicillinase produced are sufficient to account for the high degrees of resistance observed.     

Cellulase and ({beta}-Glucosidase Components in Culture Filtrates from Botryodiplodia theobromae Pat.

Cellulase and ß-glucosidase components in culturefiltrates from Botryodiplodia theobromaePat. were separatedby polyacrylamide-gel electrophoresis and gel-filtration onSephadex. A correlation between the electrophoretic mobilitiesand gel-filtration behaviour of the components was established.Four cellulase components (Ca, Cb, Cc, and Cd) were recognized with approximate molecular weights of 162 000 (Ca), 15 000(Cb), 10 000 (Cc), and 4500 (Cd). Four ß-glucosidasecomponents (Ba, Bb, Bc, and Bd) were also recognized with molecularweights of 112 000 (Ba), 56 000 (Bb), 27 000 (Be), and 13 300(Bd). The cellulase components appeared to be composed of aggregatesof similar subunits. Similarly, the ß-glucosidasecomponents appeared to be aggregates of similar subunits. Similarly,the ß-glucosidase components appeared to be aggregratesof a subunit that differed from the cellulase subunit. The activitiesof the cellulase components differed on different cellulosicsubstrates. Cotton flock was readily solubilized by a mixtureof cellulase components Ca, Cb, and Cc, an effect which wasenhanced in the presence of component Cd. Any appreciable solubilizationof native cotton fibres required component Ca or, more effectively,Cd.     

Lack of Toxicity of Helminthosporium maydis-Invaded Corn and Culture Filtrates to Chicks and Mice

Six isolates of Helminthosporium maydis, obtained from southern leaf blight-damaged corn, were grown separately on autoclaved corn and fed to chicks and mice to evaluate their toxigenicity. Two-, three-, and four-week-old culture filtrates from three pathogenic isolates grown separately on modified Fries medium were also evaluated for toxigenicity. None of the invaded corn samples or culture filtrates affected the weights of chicks or mice when compared to controls. Postmortem examinations did not reveal significant gross lesions. H. maydis was not toxigenic under our experimental conditions.