Cells of the anaerobic ruminal bacterium Fibrobacter succinogenes subsp. succinogenes S85 (formerly Bacteroides succinogenes) exhibit arylesterase activity. When cells were grown on cellulose, it was found that 69% of the total esterase activity was extracellular while 65% was nonsedimentable upon centrifugation of the culture supernatant at 100,000 x g. Disruption of the cells by various different methods failed to increase the esterase activity, indicating that the substrate was fully accessible to esterase enzymes in intact cells. During growth of cells with either glucose or cellulose as the sole carbon source, the increase in acetylesterase activity corresponded to an increase in cell density, suggesting constitutive production. The enzyme(s) hydrolyzed a-naphthyl, p-nitrophenyl, and 4-methylumbelliferyl derivatives of acetic acid; xylose tetraacetate; glucose pentaacetate; acetylxylan; and a polymer composed of ferulic acid, arabinose, and xylose in molar proportions of 1:1.1:2.2 (FAX). These data demonstrate the presence of an acetylxylan esterase and a ferulic acid esterase.

An acetylxylan esterase (EC 3.1.1.6) was purified to apparent homogeneity from the nonsedimentable extracellular culture fluid of Fibrobacter succinogenes S85 grown on cellulose. This enzyme had an apparent molecular mass of 55 kDa and an isoelectric point of 4.0. The temperature and pH optima were 45°C and 7.0, respectively. The apparent Km and V.. were 2.7 mM and 9,100 U/mg, respectively, for the hydrolysis of a-naphthyl acetate. The enzyme cleaved acetyl residues from birchwood acetylxylan but did not hydrolyze carboxymethylcellulose, larchwood xylan, ferulic acid-arabinose-xylose polymer, p-nitrophenyl-a-L-arabinofuranoside, or longer-chain naphthyl fatty acid esters. The esterase enzyme may play a role in enhancing hemicellulose degradation by F. succinogenes, thereby allowing it greater access to cellulose present in forage cell wails.

The suitability of using catheterized, partially nephrectomized (uremic) pigs to study catheter-associated infection in peritoneal dialysis was investigated. In some pigs, an inoculum of 10(9) cfu of Staphylococcus epidermidis ATCC 35984 was deposited around the catheter exit site and the organism was allowed to colonize over 21 days. The strain was recoverable from tissues and catheter samples at various locations along the catheter tract from inoculated pigs at postmortem examination. Uninoculated control catheters were colonized to a significantly lesser degree and by various other staphylococcal species. Immune response by inoculated pigs toward catheter-associated bacteria was indicated by a significant increase in serum antistaphylococcal IgG concentration and an increased percentage of peripheral polymorphonuclear leukocytes. Uremia had no significant effect on serologic response. Immunoblotting against S. epidermidis lysostaphin-extracted proteins showed that although incubation with inoculated group antisera produced more intense banding and reacted to a wider range of protein than did antisera from uninoculated controls, common antigenic proteins among the groups were found.

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Staphylococcus epidermidis RP62A (ATCC 35984) was grown in tryptic soy broth (TSB), iron-depleted TSB (TSB-Fe), iron-reconstituted TSB-Fe (TSB+Fe), a chemically defined medium, and fetal calf serum (FCS) and on silastic disks in chambers that were sutured to the pig peritoneal wall. Bacterial cell wall proteins were extracted by digestion with recombinant lysostaphin, separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and detected by silver staining. Cell wall proteins from TSB-, chemically defined medium-, or FCS-grown cells had a complex profile of greater than 25 protein bands spanning the full molecular mass range. By contrast, a digest obtained from in vivo-grown cells had only five major proteins of 40 kDa or greater. Proteins of 130 and 106 kDa were present in the cell envelopes of TSB-Feand in vivo-grown cells but not in those grown in TSB or TSB+Fe. A 43-kDa protein expressed by in vitro-grown cells and 52and 96-kDa proteins expressed by in vivo-grown cells reacted with antisera from pigs with the chamber implants and from catheterized, paracatheter-inoculated pigs but not with hyperimmune sera from pigs immunized with TSB-grown cells. The data indicate that S. epidermidis, growing under in vivo conditions, expresses antigens distinct from those that are grown in vitro.