Intravascular macrophages in lungs of pigs infected with Haemophilus pleuropneumoniae

Pigs were inoculated intratracheally with a virulent or an avirulent isolate of Haemophilus pleuropneumoniae serotype 5 and sacrificed during the first 24 hours post-inoculation. Intravascular macrophages were examined by electron microscopic and morphometric techniques. Samples of lung were taken from regions with no macroscopic lesions (Zone 0), 2.5 to 3.0 cm from lesions (Zone 1), and from the immediate edge of lesions (Zone 2). Those pigs inoculated with the avirulent isolate did not develop lesions. Pigs given the virulent isolate consistently developed necrohemorrhagic lesions in the dorsolateral aspect of the caudal and middle lung lobes. Relative volumes of intravascular macrophages in Zones 1 and 2 increased with increased time post-inoculation; in pigs given the avirulent isolate, intravascular macrophage volume decreased with increased time post-inoculation. Cytoplasmic volume to nuclear volume ratios for macrophages in Zone 2 from pigs with necrohemorrhagic lesions progressively increased with increased time post-inoculation. Enlarged intravascular macrophages had large nuclei, prominent nucleoli, and abundant cytoplasm. Increased cytoplasmic volume was the result of increased numbers of lysosomes, phagosomes, rough and smooth endoplasmic reticulum, and large Golgi complexes. Pigs inoculated with the virulent bacteria had IV macrophages with large phagosomes that contained necrotic cell debris and fibrin. Macrophages with phagosomes were more frequent in the 6-, 9-, and 24-hour sample periods of pigs with lesions than in any other group. Intercellular adhesion plaques (ICAP) were present between IV macrophages and subjacent endothelial cells. ICAP's increased in length with increased time post-inoculation in Zones 1 and 2 from pigs with necrohemorrhagic lesions. In later sample periods, multiple closely associated and interlacing IV macrophages formed a discontinuous layer over endothelial cells in Zone 2 samples near necrohemorrhagic lesions. These results suggest that the intravascular macrophage population changes from immature macrophages to mature macrophages or immature epithelioid cells within 24 hours after inhalation of a virulent Haemophilus pleuropneumoniae. Furthermore, intravascular macrophages likely function to clear cellular and acellular debris from the blood in pneumonic conditions.     

Dose response relationship of Haemophilus pleuropneumoniae aerosols in pigs.

The virulence of Haemophilus pleuropneumoniae was quantitated for ten and 12 week old pigs following aerosol exposure. The volume and concentration of culture aerosolized, the estimated numbers of organisms inhaled by the pigs and the mortality rates at 72 hours postexposure were computed and used to calculate the LD50. There was correlation between the concentration of culture aerosolized, the amount of the estimated inhaled dose and the mortality rates. The ten week old pigs were apparently more susceptible to aerosols of H. pleuropneumoniae than the 12 week old pigs. The LD50 value or a multiple of it appears to be a reasonable basis for a standardized aerosol challenge of the immunity of pigs vaccinated with experimental or commercial H. pleuropneumoniae vaccines.     

Pulmonary clearance of Haemophilus pleuropneumoniae and Serratia marcescens in mice

Bacterial counts (per lung) were made from Swiss White and/or C3H mice, inoculated intranasally with either Haemophilus pleuropneumoniae or Serratia marcescens and killed at specific times. The percentages of bacterial survival and clearance at each time were determined. Serratia marcescens was cleared progressively, but not completely, from the lungs of C3H and Swiss White mice. The overall pulmonary clearance of S marcescens by Swiss White mice was significantly greater than that of C3H mice (P less than 0.01). The pulmonary clearance pattern of H pleuropneumoniae in C3H mice differed according to the dose inoculated. With a larger H pleuropneumoniae median lethal dose (0.1 LD50), the organism multiplied consistently up to 25 times by 12 hours after the inoculations were done, when clinical signs and lesions appeared in a few of the mice. The clearance rates at 24 and 48 hours after inoculations were 60.65% and 10.3%, respectively. Mice given 0.04 LD50 H pleuropneumoniae showed a 10-fold increase of H pleuropneumoniae in the first 4 hours, with clearances reaching 33%, 66%, and 99% at 8, 12, and 24 hours, respectively.     

Incidence, economic effects, and control of Haemophilus pleuropneumoniae infections in pigs

This thesis synopsis reports the results of investigations on: the incidence of clinical and subclinical H. pleuropneumoniae infections on pig farms in North Brabant; the economic loss on fattening farms; the effect of husbandry; the possibility of eradicating H. pleuropneumoniae on infected breeding farms on the basis of serological testing; the possibility of control on fattening farms by means of vaccination. The principal findings were as follows: Clinical and subclinical H. pleuropneumoniae infections are widespread on pig farms in North Brabant: The greatest economic loss was due to mortality and veterinary fees; the daily weight again was not found to be affected. The effect of husbandry. Less mortality occurred: on farms where the animals were not shifted during the fattening period; in compartments where the all-in all-out method was systematically used; in compartments with thermostatically controlled warming of the incoming air; in compartments meeting all standards of a good housing climate. Various sanitation measures on infected breeding farms were not successful. Only one experimental vaccine afforded good protection, but because of a high percentage of injection abscesses this vaccine was unsuitable for use in practice.     

Incidence,economic effects,and control of Haemophilus pleuropneumoniae infections in pigs

Summary

This thesis synopsis reports the results of investigations on:

– the incidence of clinical and subclinical H. pleuropneumoniae infections on pig farms in North Brabant;

– the economic loss on fattening farms; the effect of husbandry;

– the possibility of eradicating H. pleuropneumoniae on infected breeding farms on the basis of serological testing;

– the possibility of control on fattening farms by means of vaccination.

The principal findings were as follows:

Clinical and subclinical H. pleuropneumoniae infections are widespread on pig farms in North Brabant:

The greatest economic loss was due to mortality and veterinary fees; the daily weight again was not found to be affected.

The effect of husbandry.

Less mortality occurred: on farms where the animals were not shifted during the fattening period; in compartments where the all‐in all‐out method was systematically used; in compartments with thermostatically controlled warming of the incoming air; in compartments meeting all standards of a good housing climate.

Various sanitation measures on infected breeding farms were not successful.

Only one experimental vaccine afforded good protection, but because of a high percentage of injection abscesses this vaccine was unsuitable for use in practice.     

Endobronchial inoculation of various doses of Haemophilus (Actinobacillus) pleuropneumoniae in pigs

Twelve-week-old specific-pathogen-free pigs were inoculated deep in the bronchi with Haemophilus (Actinobacillus) pleuropneumoniae strain 13261 in doses ranging from 8 x 10(1) to 9 x 10(7) colony-forming units (CFU). Pigs that survived infection were euthanatized and examined 48 hours after inoculation. The relationship between dose and severity of disease was evaluated clinically and the weight of pneumonic lesions was compared. The relationship between infection dose and weight of pneumonic lesions proved to be unimodal and not linear. Inoculation of 10(4) CFU of strain 13261 resulted in severe pneumonic lesions and mortality of 29%. In contrast, death was not observed after inoculation with 10(6) CFU of strain 13261 and pneumonic lesions were less severe (P less than 0.05). An infective dose of 10(3) CFU induced pneumonic lesions that tended (not statistically significant) to be less severe than those induced by a dose of 10(4) CFU. The peak fever response in all infected pigs was observed from 6 to 12 hours after inoculation. Leukocytosis developed within 12 hours after inoculation, because of an increase of neutrophilic granulocytes. Thereafter, WBC count decreased owing to lymphopenia. Serum iron concentration decreased 80% after inoculation, and zinc concentration decreased 54%.     

Haemophilus pleuropneumoniae (Actinobacillus pleuropneumoniae). Serotypes 8, 3 and 6. Serological response and cross immunity in pigs

Immunity obtained by vaccination with Haemophilus pleuropneumoniae is type specific and protection will only be obtained against the serotype contained in the vaccine. Serotype 8 is closely related to serotypes 3 and 6 and the objective of the present study was therefore to examine if cross immunity between the three serotypes could be obtained at vaccination.