ABSTRACT
Objective: Pneumonia, both in the community and the hospital setting, represents a significant cause of morbidity and mortality in the cardiothoracic patient population. Diagnosis of pneumonia can be masked by other disease processes and is often diagnosed after the patient is already experiencing the disease. A noninvasive, sensitive test for pneumonia could decrease hospitalizations and length of stay for patients. We have developed a porcine model of pneumonia and evaluated the exhaled breath of infected pigs for biomarkers of infection. Methods: Anesthetized 60-kg adult pigs were intubated, and a bronchoscope was used to instill a solution containing 12 × 108 cfu of methicillin-sensitive Staphylococcus aureus or a control solution without bacteria (Sham) into the distal airways. The pigs were then reintubated on postoperative days 3, 6, and 9, with bronchoscopic bronchial lavages taken at each time point. At each time point, a 500-mL breath was captured from each pig. The breath was evacuated over a silicon microchip, with the volatile carbonyl compounds from the breath captured via oximation reaction, and the results of this capture were analyzed by ultra-high performance liquid chromatography mass spectrometry. Results: A total of 64% of the pigs inoculated with methicillin-sensitive S. aureus demonstrated consolidation on chest radiography and increasing counts of methicillin-sensitive S. aureus in the bronchial lavages over the span of the experiment, consistent with development of pneumonia. Analysis of the exhaled breath demonstrated 1 carbonyl compound (2-pentenal) that increased 10-fold over the span of the experiment, from an average of 0.0294 nmol/L before infection to an average of 0.3836 nmol/L on postoperative day 9. The amount of 2-pentenal present was greater in the breath of infected pigs than in the noninfected pigs or the sham inoculated pigs at postoperative days 6 and 9. Using an elevated concentration of 2-pentenal as a marker of infection yielded a sensitivity of 88% and specificity of 92% at postoperative day 6, and a sensitivity and specificity of 100% at postoperative day 9. Conclusions: We were able to successfully develop a clinical pneumonia in adult 60-kg pigs. The concentration of 2-pentenal correlated with the presence of pneumonia, demonstrating the potential for this compound to function as a biomarker for methicillin-sensitive S. aureus infection in pigs.
ABSTRACT
The anther smut fungus, Microbotryum violaceum, infects over 200 species of Caryophyllaceae (Pinks). However, limited published studies, as well as anecdotal evidence, suggest that each isolate of the fungus is restricted to one or a few species that it can productively infect. In the absence of physical differences, it would be useful to have molecular markers to identify individuals with specific host ranges prior to genetic analyses of host preference. With this purpose in mind, 17 isolates from eight different host species were characterized for differences in their respective γ-tubulin genes. The region of the gene including the sixth and seventh introns and some surrounding coding regions was amplified and sequenced and the results were analyzed phylogenetically. Despite the small sample size and the geographical distribution of their respective host plants, isolates from the same host species showed no differences in the DNA regions examined; isolates of closely related pathovars also grouped together. In contrast, relative to the corresponding regions from other pathovars, isolates from host species that were genetically or taxonomically more distant showed a marked number of differences in both introns and in the third (wobble) position of codons in the seventh exon. Thus, DNA sequence differences in this highly conserved gene may be used to distinguish isolates from different host species. Such information may prove useful as markers for the different formae speciales in future analyses of host preference.
