The Outgrowth of Spores of Clostridium bifermentans

Abstract

SUMMARYGermination and outgrowth of spores of Clostridium bifermentans occurred rapidly and synchronously in a medium containing casein hydrolysate, vitamins and metal salts. Germination began immediately after inoculation and within 10 min more than 9 0 % of the spores were phase dark. Swelling of the spores began immediately thereafter and by 25 min after inoculation more than 70 % of the spores were swollen. Elongation of the swollen spores was first detected at about 35 min; by 65 min 80 % of the spores had become cells, and by 90 min cell division had started. Increases in turbidity, RNA and protein were first detected during swelling and an increase in DNA was first observed at the beginning of elongation. By the time cell division began DNA had doubled. When spores were heated at 80 "C for 10 min, 20 % survived; the survivors germinated very much more slowly than unheated spores and only formed cells if anaerobic conditions were maintained. The increases in RNA, protein and DNA occurred much later during outgrowth than with unheated spores. I N T R O D U C T I O NMany detailed studies have been made of the events which occur during the outgrowth of spores of bacilli (for a review, see Strange & Hunter, 1969). Outgrowth often occurs very rapidly, e.g. with Bacillus cereus net synthesis of RNA occurred 10 min and cell division 60 min after inoculation into growth medium (Fitz-James, 1955). With clostridial spores no comparable studies have been made. Germination, which precedes outgrowth, is often slower and less synchronous than with spores of bacilli. With Clostridium botulinum 15 % of spores were ungerminated and 45 % elongated when cell division began at 180 min, while an increase in RNA occurred after 60 min (Rowley, El-Bisi, Anellis & Snyder, I 970 ;Church, Germaine & Rowley, 1971). With such spores it is difficult to relate chemical changes to morphological development. Clostridium bifermentans has been found to germinate rapidly and synchronously (Waites &Wyatt, 1971). It was of interest, therefore, to study the morphological and chemical changes which occur during outgrowth in this strain and to compare the germination and outgrowth of heat damaged spores. METHODS

Role of the gerA operon in L-alanine germination of Bacillus licheniformisspores

“…To assay the germination response of MW3, NVH-1307 and NVH-1311 to several amino acids, casein hydrolysate was used. Casein hydrolysate consists of a mixture of amino acids made from acid hydrolyzation of the milk protein casein and has been used as a germinant for Clostridium bifermentans and B. cereus in earlier studies [61-63]. In our study, casein hydrolysate proved to be a potent germinant for B. licheniformis , giving a rapid germination response (~70% phase dark spores as visualised by phase contrast microscopy) both for the wild type MW3 and the complementation mutant NVH-1311.…”

Delayed brain abscess related to a retained foreign body with culture of Clostridium bifermentans

“…The spores swell and elongate to the vegetative phase after a period of dormancy, and begin to multiply. 24 The abscess in this case was sterilized by penicillin, metronidazole, and antibiotic irrigation of the cavity, which again suggests a lower virulence than some other species of Clostridium. The patient's records show no history of infection at his initial craniotomy or cranioplasty.…”

Microbiological Applications of the Inactivation of Antibiotics and Other Antimicrobial Agents

“…Kulikovsky et al 1975) possibly as a result of allowing germination to proceed and then killing the germinated spores. A protein may be released from the spore coat thereby producing an increased permeability of the spores to gerrninants (Wyatt & Waites 1973Waites & Wyatt 1974 (1971) stated that although halogens were unlikely to cause inhibition of grou'th in subculture media. it was important to stop the lethal action at the desired time for sampling purposes.…”