Comparison of bacterial cultivation, PCR, in situ hybridization and immunohistochemistry as tools for diagnosis of Haemophilus somnus pneumonia in cattle

The aim of the present study was to compare the potential of bacterial cultivation (BC), PCR, in situ hybridisation (ISH), and immunohistochemistry (MC) in the diagnosis of Haemophilus somnus, when applied to pneumonic bovine tissue. Lungs from 65 field cases submitted for bacteriological examination were included in the study. The PCR-detection was performed on three different samples: plate-PCR (detection on plate washes after incubation of lung tissue on agar plates); swab-PCR (direct detection on a swab from the cut surface); and, whenever possible, a bronchus-PCR (direct detection on a swab from the main bronchus of the right cranial lung lobe). In order to examine the pathological significance of the findings, a histopathological examination of the cases was performed. H. somnus was detected by one or more techniques in 33 cases in total. By BC the bacterium was isolated from 10 cases, MC and ISH were positive in 17 and 19 cases, and plate- and swab-PCR were positive in 21 and 29 cases, respectively. The bronchus-PCR was positive in 30 out of 61 cases examined. The PCR-technique was the most sensitive method, and as this technique is fast and relatively inexpensive, it should be considered as a supplementary tool in the diagnosis of H. somnus induced calf pneumonia.