Porcine reproductive and respiratory syndrome virus and Haemophilus parasuis antigen distribution in dually infected pigs.

Immunohistochemical, viral and bacterial isolation techniques were used to study the distribution and localization of porcine reproductive and respiratory syndrome virus (PRRSV) and Haemophilus (H.) parasuis in experimentally infected pigs. Thirty pigs seronegative to PRRSV and H. parasuis were divided into four groups. Group A pigs (10 animals) were inoculated with both virus and bacteria; group B pigs (10 animals) were inoculated with bacteria, group C pigs (five animals) were inoculated with virus and group D pigs (five animals) were kept as negative controls. All pigs of groups A and C became infected with PRRSV, according to virological techniques used (immunohistochemistry, virus isolation and virus serology). Lung, heart and tonsils were the most frequently immunolabeled tissues, and monocyte/macrophage lineage cells were the target for PRRSV in all tissues. All pigs in groups A and B also became infected with H. parasuis based on immunohistochemical and bacterial isolation results. Serosal surfaces, lung and tonsils were the most frequently immunolabeled tissues, and bacteria were found in monocyte/macrophage lineage cells as well as within neutrophil cytoplasm. No differences in terms of bacterial distribution or localization in tissues of pigs of groups A and B were detected. These results suggest that there is no influence of the previous infection with PRRSV in the occurrence of H. parasuis infection.

Effect of porcine reproductive and respiratory syndrome virus infection on the clearance of Haemophilus parasuis by porcine alveolar macrophages.

Porcine reproductive and respiratory syndrome virus (PRRSV) infection in young piglets is frequently associated with secondary infection due to various pathogens, especially those of the respiratory tract. One of the most important mechanisms in respiratory diseases is related to the alteration of function of porcine alveolar macrophages (PAMs). The objective of this study was to determine how PRRS virus infection affects the capabilities of PAMs in the phagocytosis and destruction of Haemophilus parasuis. Phagocytosis percentages were determined in vitro and ex vivo, after collected PAMs were directly exposed to the virus of if PAMs were collected from piglets previously infected with PRRSV. In vitro experiments demonstrated that H. parasuis uptake by PAMs is only increased in the early stages of PRRSV infection (2 h post-infection). In contrast, in the ex vivo experiments it was shown that PAMs from PRRSV-infected piglets do not seem to change in their phagocytic rate until the later stages of infection. Together with a decrease in the phagocytic rate, a marked decrease in the functional ability of PAMs to kill bacteria was observed 7 d post-infection. It is hypothesized that when animals are exposed to PRRSV, there is a marked decrease in the functional ability of PAMs to kill bacteria through the release of superoxide anion, indicating a possible negative effect of the virus, at least at the macrophage level.

Ultrastructural study of porcine alveolar macrophages infected in vitro with porcine reproductive and respiratory syndrome (PRRS) virus, with and without Haemophilus parasuis.

Two experiments were designed to study ultrastructural changes in porcine alveolar macrophages (PAM) inoculated with porcine reproductive and respiratory syndrome (PRRS) virus (experiment 1) and with PRRS virus and Haemophilus parasuis (experiment 2). In both experiments, the viral infectious dose represented a "multiplicity of infection" of 1. Viral infection alone induced minimal ultrastructural changes at this dose, consisting only of an increase in lysosome numbers. Mixed viral and bacterial infection induced the production of greatly increased numbers of phagosomes and phagolysosomes. The PAM were of low efficacy in phagocytizing H. parasuis. PRRS virus infection had only a minimal effect on the phagocytosis of H. parasuis by PAM. It is suggested that the virus induces PAM activation rather than PAM destruction.

Immunohistochemical detection of Haemophilus parasuis serovar 5 in formalin-fixed, paraffin-embedded tissues of experimentally infected swine.

An avidin-biotin complex immunohistochemistry technique was developed to detect Haemophilus parasuis serovar 5 in experimentally infected 18-21-day-old conventional pigs, using a rabbit polyclonal antiserum. Seven of 10 intratracheally inoculated animals developed a low to medium degree of fibrinous polyserositis; meninges and pleura were the most severely affected areas. Haemophilus parasuis was recovered from 9 of 10 pigs; in 2 of them H. parasuis was isolated from tracheal swabs only. Positive immunohistochemistry results, mainly observed as free bacteria or bacteria within inflammatory cell cytoplasm in the fibrinopurulent exudate, were observed in 8 of 10 animals. Cross-reactivity with Actinobacillus pleuropneumoniae was detected but not with other gram-positive and gram-negative bacteria tested. This immunohistochemistry technique seemed to be at least as sensitive as microbiologic cultures and could be useful in studies of pathogenesis and retrospective diagnosis. However, cross-reactivity with A. pleuropneumoniae means that positive immunohistochemistry results in lung tissue from field cases would be dubious.

Porcine reproductive and respiratory syndrome virus (PRRSv) interaction with Haemophilus parasuis.

The interaction of bacteria and virus has been well demonstrated in the pathogenesis of respiratory disease in swine. The interaction between porcine respiratory and reproductive syndrome virus (PRRSv) and Haemophilus parasuis has not been studied. We initiated studies to evaluate a possible effect of the PRRSv on the pathogenesis of polyserositis caused by H. parasuis. A group of 30 three week old piglets were distributed in 4 groups. Group I (10 pigs) was inoculated with PRRSv and H. parasuis. Group II (10 pigs) was inoculated with H. parasuis alone. Group III (5 pigs) was inoculated with virus alone and group IV (5 pigs) was inoculated with culture media. Lesions consisted of a severe fibrinous polyserositis affecting 7 of 10 animals in group II and a mild fibrinous pleuritis in 1 of 10 animals of group I. Three of ten animals dually infected with the two agents died during the course of the study. These animals had pulmonary congestion and focal lung hemorrhages. No other animals died from other groups. Group III and IV had no macroscopic lesions. Microscopically group III had interstitial pneumonia. Immunomodulating virus effect may explain the differences in terms of lesions severity between groups I and II. Septic shock was suspected as cause of sudden death.