The use of deep oral swabs as a surrogate for transoral tracheal wash to obtain bacterial cultures in dogs with pneumonia.

OBJECTIVE: To determine agreement in bacterial culture and susceptibility (BC&S) between deep oral swab (DOS) and routine transoral tracheal wash (TOTW) in dogs with pneumonia. DESIGN: Prospective study, performed between 2008 and 2010 with no follow-up period. SETTING: University teaching hospital. ANIMALS: Five puppies and 5 adult dogs with community or hospital-acquired pneumonia. MEASUREMENTS AND MAIN RESULTS: A DOS was collected from each patient; dogs were then briefly anesthetized with propofol(a) for a routine TOTW. Routine microbiology techniques were used for culture. Positive culture results were obtained from all 20 swabs. Results of BC&S were compared between the 2 sites. In all puppies, there was no agreement between DOS and TOTW. In adult dogs, 2 patients with hospital-acquired infection (HAI) had complete agreement between the BC&S results from DOS and TOTW. Two adult dogs had some similarity in BC&S results, and the final dog had no relationship between cultures. CONCLUSIONS: DOS is not an appropriate alternative to TOTW to obtain samples for BC&S in puppies with pneumonia. In adult dogs, traditional TOTW remains the gold standard, but DOS may be considered in dogs with suspected HAI. Further work is needed to confirm this preliminary finding.

Complex sphingolipid synthesis in plants: characterization of inositolphosphorylceramide synthase activity in bean microsomes.

Complex glycophosphosphingolipids present in plants are composed of ceramide, inositolphosphate, and diverse polar oligosaccharide substituents. The activity of inositolphosphorylceramide (IPC) synthase (phosphatidylinositol:ceramide inositolphosphate transferase), the enzyme proposed to catalyze the initial committed step in the formation of these complex sphingolipids, was characterized in wax bean hypocotyl microsomes. Enzyme activity was assayed by monitoring the incorporation of fluorescent NBD-C(6) ceramide or [3H]inositolphosphate from radiolabeled phosphatidylinositol (PI) into product identified by TLC. IPC synthase was found to utilize nonhydroxy fatty acid-containing ceramide, hydroxy fatty acid-containing ceramide, and NBD-C(6) ceramide as substrate. Maximum product formation was observed at PI concentrations in excess of 600 microM (with half-maximum activity at approximately 200 microM). Both endogenous PI and ceramide appeared to serve as substrates. Aureobasidin A and rustmicin, two potent inhibitors of fungal IPC synthase, inhibited enzyme activity in bean microsomes with values for IC(50) of 0.4-0.8 and 16-20 nM, respectively. IPC synthase activity appeared most closely associated with the Golgi based on results using selected marker enzymes. Enzyme activity was detected in a variety of plant tissues. This report, the first to characterize IPC synthase in plant tissues, demonstrates the similarities between the plant enzyme and its yeast counterpart, and provides insight into plant glycophosphosphingolipid biology.