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A polymerase chain reaction (PCR) test was developed in order to improve the accuracy and speed of diagnosis of Haemophilus parasuis, an economically important respiratory pathogen that affects swine. The gene sequence of the 16S small subunit ribosomal RNA of H. parasuis (GenBank M75065) was compared with 56 16S sequences of related bacteria, including those frequently isolated from pig tissues. Two species-specific primers were designed: HPS forward and HPS reverse. The predicted size of the amplified PCR product was 821 bp. The PCR test could detect a minimum of 102 bacteria and 0.69 pg of DNA. Thirty-one H. parasuis isolates, including 12 different serovars and 19 field isolates, were positive using the PCR test. No amplification was observed when the test was run using DNA from 15 other bacterial species commonly isolated from swine tissues. A weak band was observed when the PCR test was performed using Actinobacillus indolicus DNA as template. Clinical samples tested by PCR included tissues and swabs from 5 animals naturally infected with H. parasuis and 1 experimentally infected animal. The PCR was positive in 26 of 30 clinical samples. Four samples showed weak bands, and these results were not considered positive. Haemophilus parasuis was isolated from 18 of 30 of these samples. Tissues from specific pathogen-free (SPF) pigs and from unrelated species were negative for H. parasuis isolation and PCR. The developed PCR was successfully used in the diagnosis of H. parasuis infection, especially when compared with traditional microbiology techniques.

Immunoperoxidase assays were performed on 21 archived formalin-fixed, paraffin-embedded tissues from from American bison (Bison bison) with bronchopneumonia. Seven of the 21 bison had positive staining for Haemophilus somnus in alveolar exudate, visceral pleura, lung parenchyma, and chronic necrotic lesions, and H. somnus was isolated from tissues from 1 of these 7 animals. Results suggest that H. somnus is a respiratory pathogen in bison.

Three strains of Hemophilus somnus were infused into the posterior cervix of six pregnant cows. The organism persisted in the cervicovaginal region for eight to 87 days, and at parturition H. somnus was isolated from chorioallantois in four of six cows; placentitis developed, and fetal membranes were retained. All calves were born alive and no H. somnus was recovered from them. One cow died 14 days after parturition. The death was attributed to severe necrotizing metritis; H. somnus was not isolated from the uterus at death, but was isolated from the placenta at parturition and cervicovaginal mucus two days days later.

Eight calves were inoculated into the bronchus with H. somnus. Thirteen calves were inoculated with bovine respiratory syncytial virus (BRSV) and 8 days later with H. somnus. All calves developed necrotizing, suppurative, lobular bronchopneumonia and pleuritis. Clinical signs of disease and pneumonic lesions were significantly more severe in calves that were sequentially inoculated with BRSV followed by H. somnus. Pneumonic lesions in the inoculated calves were similar to those described for naturally occurring H. somnus-associated respiratory tract disease. Control calves inoculated with BRSV alone or sham-inoculated with medium did not develop clinical signs of respiratory tract disease. The BRSV-inoculated control calves developed minimal pneumonic lesions.

Hemophilus parainfluenzae may be a normal inhabitant of the pharynx, but it can have pathogenic potential elsewhere in the body. It has been identified as a cause of bacteremia and meningitis. The case discussed here represents the first report of H parainfluenzae-induced supraglottitis in a child. In reviewing its pathogenic potential in the upper respiratory tract, it appears that its major effect could be the ability to transfer ampicillin resistance to ampicillin-susceptible Hemophilus influenzae.

The injection of three strains of Hemophilus somnus into the amniotic cavity of pregnant cows produced death of the fetus and abortion followed by retention of the placenta. H. somnus was recovered readily from the placenta but less so from the fetus. Lesions in the placenta were conspicuous in arteries and somewhat resembled vascular lesions observed in septicemic H. somnus. H. somnus was recovered from the maternal circulation in several infected cows. Clinical signs included fever, irritability, lameness, and vulvar discharge, but no cows died. The organism was recovered for prolonged periods in cervicovaginal mucus and urine after remission of clinical signs. This may be a means of spread of infection.