Skewed genomic variability in strains of the toxigenic bacterial pathogen, Clostridium perfringens

Clostridium perfringens is a Gram-positive, anaerobic spore-forming bacterium commonly found in soil, sediments, and the human gastrointestinal tract. C. perfringens is responsible for a wide spectrum of disease, including food poisoning, gas gangrene (clostridial myonecrosis), enteritis necroticans, and non-foodborne gastrointestinal infections. The complete genome sequences of Clostridium perfringens strain ATCC 13124, a gas gangrene isolate and the species type strain, and the enterotoxin-producing food poisoning strain SM101, were determined and compared with the published C. perfringens strain 13 genome. Comparison of the three genomes revealed considerable genomic diversity with >300 unique "genomic islands" identified, with the majority of these islands unusually clustered on one replichore. PCR-based analysis indicated that the large genomic islands are widely variable across a large collection of C. perfringens strains. These islands encode genes that correlate to differences in virulence and phenotypic characteristics of these strains. Significant differences between the strains include numerous novel mobile elements and genes encoding metabolic capabilities, strain-specific extracellular polysaccharide capsule, sporulation factors, toxins, and other secreted enzymes, providing substantial insight into this medically important bacterial pathogen. ©2006 by Cold Spring Harbor Laboratory Press

Organization of the cpe Locus in CPE-Positive Clostridium perfringens Type C and D Isolates

Clostridium perfringens enterotoxin (encoded by the cpe gene) contributes to several important human, and possibly veterinary, enteric diseases. The current study investigated whether cpe locus organization in type C or D isolates resembles one of the three (one chromosomal and two plasmid-borne) cpe loci commonly found amongst type A isolates. Multiplex PCR assays capable of detecting sequences in those type A cpe loci failed to amplify products from cpe-positive type C and D isolates, indicating these isolates possess different cpe locus arrangements. Therefore, restriction fragments containing the cpe gene were cloned and sequenced from two type C isolates and one type D isolate. The obtained cpe locus sequences were then used to construct an overlapping PCR assay to assess cpe locus diversity amongst other cpe-positive type C and D isolates. All seven surveyed cpe-positive type C isolates had a plasmid-borne cpe locus partially resembling the cpe locus of type A isolates carrying a chromosomal cpe gene. In contrast, all eight type D isolates shared the same plasmid-borne cpe locus, which differed substantially from the cpe locus present in other C. perfringens by containing two copies of an ORF with 67% identity to a transposase gene (COG4644) found in Tn1546, but not previously associated with the cpe gene. These results identify greater diversity amongst cpe locus organization than previously appreciated, providing new insights into cpe locus evolution. Finally, evidence for cpe gene mobilization was found for both type C and D isolates, which could explain their cpe plasmid diversity

Influence of environmental calcium/phosphate and pH on glass ionomers

This study investigated the effects of environmental calcium/phosphate and pH on the hardness and elastic modulus of two glass-ionomer cements (GICs) [Fuji IX Fast (FN) and KetacMolar (KM)]. Specimens were randomly subjected to storage media of pH 3, 5, and 7. The calcium and phosphate levels of the storage solutions ranged from 0 to 2.4 mM. After 4 wk of conditioning, hardness and elastic modulus were determined using a depth-sensing microindentation test. Sectioned surfaces were observed with scanning electron microscopy. For both FN and KM, no significant change in hardness, elastic modulus, or surface structure were observed at pH 7 and 5, regardless of the concentration of calcium and phosphate. FN and KM specimens conditioned at pH 3 had lower hardness and modulus in comparison to those conditioned at pH 7. An increased level of environmental phosphate led to higher hardness and elastic modulus of FN and KM at pH 3. In general, a microscopic surface reaction layer was observed in specimens conditioned at pH 3. The thickness and structure of the reaction layer varied depending on environmental phosphate levels. The results suggest that the influence of environmental calcium and phosphate on GICs was pH dependent