Pulmonary lesions induced by a Pasteurella haemolytica cytotoxin preparation in calves

Intrabronchial instillation of a Pasteurella haemolytica type A1 crude cytotoxin preparation in calves resulted in pulmonary gross and microscopic lesions comparable to spontaneous and experimental pasteurellosis. In the acute stage of the lesion electronmicroscopy revealed intravascular accumulation, degeneration and fragmentation of leukocytes in the interalveolar septa. Secondary thrombus formation and increased vascular permeability resulted in alveolar flooding, fibrin deposition, extravasation of erythrocytes and loss of alveolar epithelium. No cytotoxicity was observed for the tracheal (in vitro) and bronchial epithelium (in vivo). The pathogenesis of the vascular lesions and their significance for the development of the typical lesions of pneumonic pasteurellosis is discussed.     

Pulmonary lesions induced by Pasteurella haemolytica in neutrophil sufficient and neutrophil deficient calves.

The role of neutrophils in the development of peracute lung lesions of bovine pneumonic pasteurellosis was investigated. Eight calves were divided into two groups of four calves each. Group I was treated with intravenous phosphate-buffered saline and served as the neutrophil sufficient calves. Group II was treated with intravenous hydroxyurea which produced a state of neutropenia. When peripheral blood neutrophil numbers dropped below 300 cells/microL in group II, all calves were challenged with an intrabronchial bolus of Pasteurella haemolytica in the log phase of growth. An acute inflammatory process occurred in both groups of calves indicated by a rise in body temperature. While pulmonary lesions occurred in both groups by six hours postinoculation, they varied in pathological characteristics. Pulmonary lesions in the neutrophil sufficient calves consisted of fibrinopurulent alveolitis-bronchiolitis with associated alveolar septal necrosis, interlobular edema, and intravascular thrombi. The neutrophil deficient calves had extensive intra-alveolar edema, interlobular edema, intraalveolar hemorrhage, atelectasis, and focal areas of alveolar septal necrosis. These results show that P. haemolytica can induce severe pulmonary tissue damage through both neutrophil dependent and neutrophil independent mechanisms.     

牛源溶血性巴氏杆菌的分离鉴定

乌鲁木齐某肉牛养殖场育肥牛发生了以体温升高、呼吸困难、咳嗽及快速死亡为主要特征的疾病,每日死亡率超过5%。从发病死亡牛肝脏和肺脏等组织中分离到1株细菌,通过菌落形态、染色特性、培养特性及生化试验等一系列鉴定,确定为溶血性巴氏杆菌。给小白鼠腹腔注射该菌悬液,8 h内全部死亡,结合该批次牛刚从伊犁长途运输而来的流行病学特点,诊断该病是以溶血性巴氏杆菌为主要病原的牛运输热。药敏试验结果表明沙拉沙星和强力霉素最敏感,将它们用于全群预防和治疗后效果显著。     

Cell-mediated Hypersensitivity to Pasteurella hemolytica in Normal and Cold-stressed Mice

Mice infected with Pasteurella hemolytica developed cell-mediated hypersensitivity manifested by inhibition of macrophage migration but not by delayed skin reactions, to two antigen preparations from this organism. Relative to migration by the cells of uninfected animals, migration by cells from infected animals was inhibited to the same degree whether or not the animals were chilled. However, the cells from infected chilled animals were not able to migrate as far as the cells from normal mice even in the absence of antigen.     

Recovery of Pasteurella hemolytica from aerosols at differing temperature and humidity.

A Pasteurella hemolytica suspension with fetal calf serum was aerosolized in a standard system with ambient temperature of 30 or 2 degrees C and relative humidity conditions of 90 or 60%. The number of organisms sprayed in five minutes and the number recovered from one third of the aerosol during these five minutes was determined. Recoveries were influenced by temperature difference between aerosol and collecting fluid. Recoveries ranged between 0.059--0.94%.     

Immunoperoxidase evaluation of pneumonic lesions induced by Pasteurella multocida in calves

To evaluate the relationship between pneumonic lesions and distribution of bacteria, lungs from calves inoculated with Pasteurella multocida were examined histologically by use of immunoperoxidase technique. Pneumonic lesions fundamentally consisted of broncho-pneumonia with fibrinopurulent pleuritis. The lesions were confirmed to be associated with inoculated P multocida, using the immunoperoxidase technique. The P multocida antigen was detected not only in the bacterial clusters in the lesions, but also in the cytoplasm of infiltrating neutrophils and macrophages. Further, immunoelectron microscopy indicated that the inoculated bacteria generally were phagocytosed and digested by neutrophils.     

Immunoperoxidase evaluation of pneumonic lesions induced by Pasteurella haemolytica in calves

Calves inoculated with Pasteurella haemolytica serovar 1 developed lesions of coagulation necrosis in the lungs that were sharply demarcated by leukocytes. The P haemolytica antigen was detected in the area of coagulation necrosis in histologic sections, using an immunoperoxidase technique. In the central area of the necrotic tissue, the bacterial antigen was diffusely presented in the necrotic alveolar wall, fibrin, serous exudate, and degenerated leukocyte. The bacterial antigen also was found in some groups of degenerating leukocytes around the necrotic tissue. The bacterial colonies among these leukocytes had strong specific reactions against P haemolytica. The bacterial antigen was observed in the cytoplasm of macrophages in alveoli around the necrotic lesion. These findings confirmed that coagulation necrosis is an important lesion in calves with pneumonia caused by P haemolytica.     

The pulmonary clearance of Pasteurella hemolytica in calves infected with bovine parainfluenza-3 virus.

The purpose of this study was to determine if parainfluenza-3 virus in calves interfered with normal pulmonary bacterial clearance. Three groups (A, B and C) of four calves each were exposed to an aerosol of parainfluenza-3 virus. Three days later the four hour pulmonary clearance of Pasteurella hemolytica was determined on the first group (A), seven days later on the second (B) and 11 days later on the third (C). Group A had a mean pulmonary bacterial retention of 3.6 +/- 3.5%. Group B was 83.1 +/- 35.9% and Group C was 41.2 +/- 30.9%. The results demonstrate that parainfluenza-3 virus interfered with the pulmonary bacterial clearance of Pasteurella hemolytica particularly on day 7 and also on day 11 but not on day 3. This inhibition of pulmonary clearance caused by the virus may be a key factor in the pathogenesis of pneumonic pasteurellosis. Histological examination of the lungs did not demonstrate a correlation between pulmonary retention of bacteria and the development of pathological changes.     

Multiple drug resistance in Pasteurella multocida and Pasteurella haemolytica from cattle and swine.

The results of antimicrobial susceptibility testing on 262 strains of Pasteurella multocida and 141 strains of Pasteurella haemolytica isolated from cattle and swine from 1971 to 1974 were analyzed for patterns of resistance to streptomycin, penicillin, tetracycline, and chloramphenicol, using a modified Kirby-Bauer procedure. Resistance was recorded for 80.5% of the isolants of P multocida and 92.2% of those of P haemolytica. Resistance to streptomycin was most frequent, followed by resistance to penicillin and tetracycline. Most cultures of P multocida and P haemolytica were susceptible to chloramphenicol. There were 9 patterns of resistance with the aforementioned antibiotics. The combinations, streptomycin and penicillin and streptomycin and tetracycline, each accounted for approximately 10% of the resistance patterns of P multocida. Approximately half of the 14 isolants of P haemolytica were resistant to the combination of streptomycin, penicillin, and tetracycline. These observations underscore the need for antimicrobial susceptibility testing of clinical isolants of P multocida and P haemolytica.     

Pulmonary response to intratracheal challenge with Pasteurella haemolytica and Pasteurella multocida.

Calves were inoculated intratracheally with 5 X 10(7), 5 X 10(8), or 5 X 10(9) colony forming units of either 18-hour stationary phase cultures or 4-hour log phase cultures of Pasteurella haemolytica. The log phase culture at all concentrations produced more severe clinical signs, hematological changes and pulmonary lesions at postmortem examination than did the corresponding stationary phase culture. More severe effects were seen with the larger doses especially with the log phase culture. Fibrinous bronchopneumonia with focal or multifocal necrosis was consistently produced by both the stationary and log phase cultures. To determine if this lesion was peculiar to P. haemolytica or whether it could be produced generally by rapidly growing Gram negative organisms, a 4-hour log phase culture of Pasteurella multocida was prepared in an identical manner to that used for the culture of P. haemolytica and given to calves intratracheally at the high bacterial dose (5 X 10(9]. The P. haemolytica produced more severe clinical, hematological and morphological changes than did the P. multocida. The lesions observed with P. multocida differed morphologically from those of P. haemolytica; there was a suppurative exudative component and minimal to no necrosis with P. multocida. It appears that an important pathogenic principle is produced by the rapidly growing P. haemolytica that causes it to produce a more severe clinical disease and more necrotizing pulmonary lesions than P. multocida.     

Serotype-specific resistance to nasal colonization by Pasteurella haemolytica in cattle

Vaccination of calves with a Pasteurella haemolytica serotype 1 antigen preparation elicited a serotype-specific inhibition of nasal colonization by P haemolytica under field conditions. Inhibition was evidenced by a low frequency of nasal colonizations and by relatively few P haemolytica serotype 1 organisms isolated from vaccinated calves. The study comprised 3 field trials, each on a separate year, and included 480 calves.     

Pulmonary lesions induced by 3-methylindole and bovine respiratory syncytial virus in calves

Our objectives were to describe the ultrastructural morphogenesis of pulmonary lesions induced by 3-methylindole in 30- to 45-day-old Holstein calves and to determine whether toxic exposure to 3-methylindole exacerbates pulmonary lesions induced by bovine respiratory syncytial virus. Administration of 3-methylindole (0.25 g/kg) to calves resulted in interstitial edema and ultrastructural swelling of type-I alveolar epithelial cells and nonciliated bronchiolar epithelial cells as early as 4 to 6 hours after intraruminal administration. More severe alveolar edema containing protein was associated with swelling of capillary endothelial cells at 2 days after administration. Proliferation of type-II alveolar epithelial cells was first observed at 2 days after 3-methylindole administration, and marked hyperplasia of type-II epithelial cells and nonciliated bronchiolar epithelial cells was evident by 4 days after administration. Pulmonary cytochrome P-450 monooxygenase concentrations decreased significantly (P less than 0.001) by 12 hours after administration and did not increase significantly again by 8 days after administration. Calves were inoculated with bovine respiratory syncytial virus 3 days after administration of 3-methylindole, and pulmonary lesions were assessed 5 days after viral inoculation. Viral replication was demonstrated by fluorescence microscopy for viral antigen or by transmission electron microscopy in ciliated and nonciliated airway epithelial cells. Viral antigen was identified infrequently in alveolar macrophages and in type-II alveolar epithelial cells. 3-Methylindole exposure in calves did not result in more widespread distribution of viral antigen in alveolar tissue of respiratory syncytial virus-inoculated calves or in significant enhancement of viral pneumonia.(ABSTRACT TRUNCATED AT 250 WORDS)     

Ibuprofen prevents Pasteurella hemolytica endotoxin-induced changes in plasma prostanoids and serotonin, and fever in sheep

Intravenous infusion of Pasteurella hemolytica endotoxin caused marked increases in the plasma levels of thromboxane B2 (TxB2), prostaglandins (PG) and serotonin in sheep. The control values for TxB2, 6-keto-PGF1 alpha, PGF2 alpha, and serotonin before endotoxin infusion averaged 283 +/- 53 (standard error of mean), 281 +/- 14 and 199 +/- 27 pg/ml and 57 +/- 3 ng/ml, respectively. At 50 min during endotoxin infusion, these values were increased to their maximum of 376, 339, 325 and 202% of control, respectively. Body temperature increased from the control value of 39.5 +/- 0.1 degrees C to a maximum of 41.5 +/- 0.1 degrees C at 200-300 min of infusion. In the second part of this study, we have examined the effects of ibuprofen on endotoxin-induced increases in plasma PG, TxB2, and serotonin levels and body temperature. The control values for TxB2, 6-keto-PGF1 alpha, PGF2 alpha, and temperature prior to ibuprofen and endotoxin infusion averaged 238 +/- 35, 335 +/- 44 and 248 +/- 28 pg/ml, 65 +/- 3 ng/ml and 40.1 +/- 0.2 degrees C, respectively. A loading dose (15 mg/kg) of ibuprofen was followed by infusion of endotoxin (12 micrograms/kg) and ibuprofen (43.3 mg/kg) over 500 min. Plasma levels of 6-keto-PGF1 alpha and serotonin increased only to 131 and 149% of control at 50 min of infusion, and levels of PGF2 alpha and TxB2 decreased to 50 and 80% of control at 100 and 150 min of infusion, respectively. Temperature remained unchanged. Ibuprofen effectively suppressed endotoxin-induced increases in the plasma levels of TxB2, 6-keto-PGF1 alpha, PGF2 alpha, and serotonin and body temperature. It was concluded from the present study that nonsteroidal anti-inflammatory drugs as an adjunct to antibiotic therapy might have a rational basis in treatment of endotoxin toxicity.     

Selenium effects on glutathione peroxidase and the immune response of stressed calves challenged with Pasteurella hemolytica

The present study was conducted to determine whether a marginal Se deficiency affects health, blood characteristics and the immune response of calves subjected to stresses associated with weaning, shipping (332 km) and Pasteurella hemolytica inoculation. Treatments were 1) -Se, 2) -Se/P. hemolytica, 3) +Se (.1 mg Se/kg feed) and 4) +Se/P. hemolytica. Previous Se intake was controlled; dams of -Se calves were fed diets marginally deficient in Se (.03 to .05 mg/kg), whereas dams of +Se calves received a s.c. injection of 30 mg Se (as sodium selenite) every 60 d. Calves were inoculated with P. hemolytica intratracheally on d 3 following weaning and transport. Inoculation with P. hemolytica increased (P less than .05) body temperatures, platelet counts, serum IgM concentrations and serum antibody titers and decreased serum albumin concentrations at 4 to 7 d postinoculation. Weight gains for the 21-d study were not affected by Se status, although whole blood and plasma glutathione peroxidase (GSH-Px) were higher (P less than .05) for +Se calves. Plasma GSH-Px increased (P less than .01) in calves showing signs of morbidity. Increases in plasma GSH-Px were correlated positively with body temperature. Serum IgM concentrations were higher (P less than .05) in +Se calves on d 17, but Se-supplemented calves had lower (P less than .05) anti-P. hemolytica titers on d 17 than -Se calves. Selenium status did not affect body temperatures, plasma creatine phosphokinase or serum IgG and albumin concentrations. These results indicate that Se status can affect IgM concentrations following stress.