Reaction of Bacillus subtilis products with amebocyte lysates of the Japanese horseshoe crab, Tachypleus tridentatus. 1988

D S Hodes, and E J Bottone, and A Hass, and H L Hodes
Department of Pediatrics, Mount Sinai School of Medicine, City University of New York, New York 10029.

Autoclaved aqueous extracts of Candida albicans cells (and the glucans isolated from them) give a positive reaction with a chromogenic substrate combined with amebocyte lysates of the Japanese horseshoe crab, Tachypleus tridentatus (CS-TAL). The extracts and glucans activate the lysate enzyme compound G, which in turn activates clotting enzyme. Activated clotting enzyme causes a positive CS-TAL reaction. C. albicans extracts and glucans react positively with a commercially available, unaltered CS-TAL preparation (Toxicolor), but they give a negative reaction with a CS-TAL from which compound G has been excluded (Endospecy). An autoclaved, sterile preparation of Sabouraud glucose broth used as a control in one experiment gave (like Candida extracts) a positive reaction with Toxicolor and a negative reaction with Endospecy. We found that the peptone powder used to make the Sabouraud glucose broth was contaminated with a strain of Bacillus subtilis. Autoclaved aqueous extracts of culture-grown B. subtilis cells were positive with Toxicolor and negative with Endospecy. This was also the case with two other strains of B. subtilis. Polysaccharides obtained from these extracts gave the same result. Endotoxin activates clotting enzyme through activation of the lysate enzyme compound C, which is present in both Toxicolor and Endospecy. Endotoxin, therefore, reacts with both CS-TAL preparations. Simultaneous assay with Toxicolor and Endospecy distinguishes endotoxin from fungal products, but since products of fungi and B. subtilis both give a positive Toxicolor and a negative Endospecy test, a simultaneous assay cannot differentiate them. However, this does not decrease the clinical value of the simultaneous Toxicolor-Endospecy assay for distinguishing fungal infection from endotoxemia because B. subtilis so rarely causes disease that it can be excluded from clinical consideration.

UI MeSH Term Description Entries
D008033 Limulus Test Sensitive method for detection of bacterial endotoxins and endotoxin-like substances that depends on the in vitro gelation of Limulus amebocyte lysate (LAL), prepared from the circulating blood (amebocytes) of the horseshoe crab, by the endotoxin or related compound. Used for detection of endotoxin in body fluids and parenteral pharmaceuticals. Limulus Tests,Test, Limulus,Tests, Limulus
D010450 Endopeptidases A subclass of PEPTIDE HYDROLASES that catalyze the internal cleavage of PEPTIDES or PROTEINS. Endopeptidase,Peptide Peptidohydrolases
D002176 Candida albicans A unicellular budding fungus which is the principal pathogenic species causing CANDIDIASIS (moniliasis). Candida albicans var. stellatoidea,Candida stellatoidea,Dematium albicans,Monilia albicans,Myceloblastanon albicans,Mycotorula albicans,Parasaccharomyces albicans,Procandida albicans,Procandida stellatoidea,Saccharomyces albicans,Syringospora albicans
D003470 Culture Media Any liquid or solid preparation made specifically for the growth, storage, or transport of microorganisms or other types of cells. The variety of media that exist allow for the culturing of specific microorganisms and cell types, such as differential media, selective media, test media, and defined media. Solid media consist of liquid media that have been solidified with an agent such as AGAR or GELATIN. Media, Culture
D004731 Endotoxins Toxins closely associated with the living cytoplasm or cell wall of certain microorganisms, which do not readily diffuse into the culture medium, but are released upon lysis of the cells. Endotoxin
D005936 Glucans Polysaccharides composed of repeating glucose units. They can consist of branched or unbranched chains in any linkages. Glucan,Polyglucose,Polyglucoses,Glucan (BO),Glucose Polymer,Polycose,Polymer, Glucose
D006737 Horseshoe Crabs An arthropod subclass (Xiphosura) comprising the North American (Limulus) and Asiatic (Tachypleus) genera of horseshoe crabs. Crabs, Horseshoe,Limulus,Limulus polyphemus,Tachypleus,Xiphosura,Crab, Horseshoe,Horseshoe Crab,Xiphosuras
D000818 Animals Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA. Animal,Metazoa,Animalia
D001412 Bacillus subtilis A species of gram-positive bacteria that is a common soil and water saprophyte. Natto Bacteria,Bacillus subtilis (natto),Bacillus subtilis subsp. natto,Bacillus subtilis var. natto

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