Mannheimia haemolytica: bacterial-host interactions in bovine pneumonia.

Mannheimia haemolytica serotype S1 is considered the predominant cause of bovine pneumonic pasteurellosis, or shipping fever. Various virulence factors allow M haemolytica to colonize the lungs and establish infection. These virulence factors include leukotoxin (LKT), lipopolysaccharide, adhesins, capsule, outer membrane proteins, and various proteases. The effects of LKT are species specific for ruminants, which stem from its unique interaction with the bovine ß2 integrin receptor present on leukocytes. At low concentration, LKT can activate bovine leukocytes to undergo respiratory burst and degranulation and stimulate cytokine release from macrophages and histamine release from mast cells. At higher concentration, LKT induces formation of transmembrane pores and subsequent oncotic cell necrosis. The interaction of LKT with leukocytes is followed by activation of these leukocytes to undergo oxidative burst and release proinflammatory cytokines such as interleukins 1, 6, and 8 and tumor necrosis factor α. Tumor necrosis factor α and other proinflammatory cytokines contribute to the accumulation of leukocytes in the lung. Formation of transmembrane pores and subsequent cytolysis of activated leukocytes possibly cause leakage of products of respiratory burst and other inflammatory mediators into the surrounding pulmonary parenchyma and so give rise to fibrinous and necrotizing lobar pneumonia. The effects of LKT are enhanced by lipopolysaccharide, which is associated with the release of proinflammatory cytokines from the leukocytes, activation of complement and coagulation cascade, and cell cytolysis. Similarly, adhesins, capsule, outer membrane proteins, and proteases assist in pulmonary colonization, evasion of immune response, and establishment of the infection. This review focuses on the roles of these virulence factors in the pathogenesis of shipping fever.

Potential involvement of gelatinases and their inhibitors in Mannheimia haemolytica pneumonia in cattle.

Mannheimia haemolytica infection of the lower respiratory tract of cattle results in a bronchofibrinous pneumonia characterized by massive cellular influx and lung tissue remodeling and scarring. Since altered levels of gelatinases and their inhibitors have been detected in a variety of inflammatory conditions and are associated with tissue remodeling, we examined the presence of gelatinases in lesional and nonlesional lung tissue obtained from calves experimentally infected with M. haemolytica. Lesional tissue had elevated levels of progelatinase A and B and active gelatinase A and B when compared with nonlesional tissue obtained from the same lung lobe. In vitro, M. haemolytica products stimulated production of gelatinase B, but not its activation, by bovine monocytes. Alveolar macrophages showed constitutive production of gelatinase B but no change in response to M. haemolytica products. Bovine neutrophils exposed to M. haemolytica products also released gelatinase B, and there was a significant increase in the activated form of this enzyme. These effects were virtually identical when recombinant O-sialoglycoprotease was used to stimulate these cells. M. haemolytica products also enhanced the expression by bovine monocytes and alveolar macrophages of the tissue inhibitor of metalloproteinase 1. Our results provide evidence that matrix metalloproteinases are activated in lung lesions from cattle with shipping fever and that M. haemolytica virulence products induce production, release, and especially activation of gelatinase B by bovine inflammatory cells in vitro.

Effect of danofloxacin and tilmicosin on body temperatures of beef calves with pneumonia experimentally induced by inoculation with Mannheimia haemolytica.

OBJECTIVE: To examine effects of danofloxacin and tilmicosin on continuously recorded body temperature in beef calves with pneumonia experimentally induced by inoculation of Mannheimia haemolytica. ANIMALS: 41 Angus-cross heifers (body weight, 160 to 220 kg) without a recent history of respiratory tract disease or antimicrobial treatment, all from a single ranch. PROCEDURE: Radiotransmitters were implanted intravaginally in each calf. Pneumonia was induced intrabronchially by use of logarithmic-phase cultures of M. haemolytica. At 21 hours after inoculation, calves were treated with saline (0.9% NaCl) solution, danofloxacin, or tilmicosin. Body temperature was monitored from 66 hours before inoculation until 72 hours after treatment. Area under the curve (AUC) of the temperature-time plot and mean temperature were calculated for 3-hour intervals and compared among treatment groups. RESULTS: The AUCs for 3-hour intervals did not differ significantly among treatment groups for any of the time periods. Analysis of the mean temperature for 3-hour intervals revealed significantly higher temperatures at most time periods for saline-treated calves, compared with temperatures for antimicrobial-treated calves; however, we did not detect significant differences between the danofloxacin- and tilmicosin-treated calves. The circadian rhythm of temperatures before exposure was detected again approximately 48 hours after bacterial inoculation. CONCLUSIONS AND CLINICAL RELEVANCE: Danofloxacin and tilmicosin did not differ in their effect on mean body temperature for 3-hour intervals but significantly decreased body temperature, compared with body temperature in saline-treated calves. Normal daily variation in body temperature must be considered in the face of respiratory tract disease during clinical evaluation of feedlot cattle.

Effects of the synthetic selectin inhibitor TBC1269 on tissue damage during acute Mannheimia haemolytica-induced pneumonia in neonatal calves.

OBJECTIVE: To determine effects of the selectin inhibitor TBC1269 on neutrophil-mediated pulmonary damage during acute Mannheimia haemolytica-induced pneumonia in newborn calves. ANIMALS: Eighteen 1- to 3-day-old colostrum-deprived calves. PROCEDURE: Mannheimia haemolytica or saline (0.9% NaCl) solution was inoculated in both cranial lung lobes of 12 and 6 calves, respectively. Calves were euthanatized 2 (saline, n = 3; M haemolytica, n = 4) or 6 hours (saline, n = 3; M haemolytica, n = 8) after inoculation. Four M haemolytica-inoculated calves euthanatized at 6 hours also received TBC1269 (25 mg/kg, IV) 30 minutes before and 2 hours after inoculation. Conjugated diene (CD) concentrations, inducible nitric oxide synthase (iNOS) expression, and apoptotic cell counts were determined in lung specimens collected during necropsy. RESULTS: Conjugated diene concentrations were significantly increased in all M haemolytica-inoculated groups, compared with saline-inoculated groups. Calves treated with TBC1269 had decreased concentrations of CD, compared with untreated calves, although the difference was not significant. Number of apoptotic neutrophils and macrophages increased significantly inTBC1269-treated calves, compared with untreated calves. Inducible nitric oxide synthase was expressed by epithelial cells and leukocytes. However, iNOS was less abundant in airway epithelial cells associated with inflammatory exudates. Degree of iNOS expression was similar between TBC1269-treated and untreated calves. CONCLUSIONS: Mannheimia haemolytica infection in neonatal calves resulted in pulmonary tissue damage and decreased epithelial cell iNOS expression. The selectin inhibitor TCB1269 altered, but did not completely inhibit, neutrophil-mediated pulmonary damage.

Coronavirus and Pasteurella infections in bovine shipping fever pneumonia and Evans' criteria for causation.

Respiratory tract infections with viruses and Pasteurella spp. were determined sequentially among 26 cattle that died during two severe epizootics of shipping fever pneumonia. Nasal swab and serum samples were collected prior to onset of the epizootics, during disease progression, and after death, when necropsies were performed and lung samples were collected. Eighteen normal control cattle also were sampled at the beginning of the epizootics as well as at weekly intervals for 4 weeks. Respiratory bovine coronaviruses (RBCV) were isolated from nasal secretions of 21 and 25 cattle before and after transport. Two and 17 cattle nasally shed Pasteurella spp. before and after transport, respectively. RBCV were isolated at titers of 1 x 10(3) to 1.2 x 10(7) PFU per g of lung tissue from 18 cattle that died within 7 days of the epizootics, but not from the lungs of the remaining cattle that died on days 9 to 36. Twenty-five of the 26 lung samples were positive for Pasteurella spp., and their CFU ranged between 4.0 x 10(5) and 2.3 x 10(9) per g. Acute and subacute exudative, necrotizing lobar pneumonia characterized the lung lesions of these cattle with a majority of pneumonic lung lobes exhibiting fibronecrotic and exudative changes typical of pneumonic pasteurellosis, but other lung lobules had histological changes consisting of bronchiolitis and alveolitis typical of virus-induced changes. These cattle were immunologically naive to both infectious agents at the onset of the epizootics, but those that died after day 7 had rising antibody titers against RBCV and Pasteurella haemolytica. In contrast, the 18 clinically normal and RBCV isolation-negative cattle had high hemagglutinin inhibition antibody titers to RBCV from the beginning, while their antibody responses to P. haemolytica antigens were delayed. Evans' criteria for causation were applied to our findings because of the multifactorial nature of shipping fever pneumonia. This analysis identified RBCV as the primary inciting cause in these two epizootics. These viruses were previously not recognized as a causative agent in this complex respiratory tract disease of cattle.

Ultrastructural characterization of apoptosis in bovine lymphocytes exposed to Pasteurella haemolytica leukotoxin.

OBJECTIVE: To characterize ultrastructural changes of bovine lymphocytes exposed to Pasteurella haemolytica leukotoxin (LKT). SAMPLE POPULATION: Partially purified LKT from a wild type P. haemolytica A1 strain and inactive pro-LKT from an isogeneic mutant Phaemolytica strain. Isolated bovine lymphocytes were obtained from 2 healthy calves. PROCEDURE: Isolated bovine lymphocytes were incubated with various concentrations of LKT and pro-LKT for 3 hours at 37 C and examined by use of transmission electron microscopy. A cytochemical Klenow DNA fragmentation assay was used to examine lymphocytes for DNA fragmentation. RESULTS: Lymphocytes incubated with LKT at a high concentration (1.0 toxic U/ml) had ultrastructural evidence of cytoplasmic and nuclear membrane rupture and swelling or lysis of mitochondria. Low concentrations of leukotoxin (0.1 toxic U/ml) induced DNA fragmentation in 80% of lymphocytes. Ultrastructurally, these cells had nuclear membrane blebbing, cytoplasmic vaculation, chromatin condensation, nuclear fragmentation, and membrane-bound apoptotic bodies. Incubation of lymphocytes with LKT at extremely low concentrations (0.001 toxic U/ml) or with pro-LKT did not alter their ultrastructure. Inclusion of 0.5 mM ZnCl2 in the medium blocked leukotoxin-induced ultrastructural changes in bovine lymphocytes. CONCLUSIONS AND CLINICAL RELEVANCE: Low concentrations of LKT induce apoptosis and high concentrations induce oncotic cell lysis in bovine lymphocytes. The ability of low LKT concentrations to induce apoptosis in host leukocytes may allow bacteria to escape host immune surveillance and colonize the host.

Comparison of serologic and protective responses induced by two Pasteurella vaccines.

Vaccine development for the prevention of pneumonic pasteurellosis remains a critical issue for the feedlot industry. Most currently available Pasteurella vaccines are formulated to stimulate immunity by either providing an adequate antigenic mass in the administered dose, or by relying on subsequent production of antigens by in vivo growth of live organisms. The ability of these different types of vaccines to stimulate rapid and high titres to key antigens is a key factor that will influence subsequent resistance to disease. The serologic and protective responses to a streptomycin-dependent, modified-live vaccine and a killed (bacterin-toxoid) vaccine against experimental pneumonic pasteurellosis were compared. Calves were vaccinated with a single injection of either a test vaccine or phosphate-buffered saline, challenged 14 d later by transthoracic injection with Pasteurella haemolytica, and euthanized 3 d post-challenge to evaluate the severity of pneumonia. On days 0, 7, and 14, serologic responses to various P. haemolytica antigens, including cell-associated and soluble antigens, were determined by enzyme-linked immunosorbent assays, and anti-leukotoxin antibody levels were determined by leukotoxin neutralization. The bacterin-toxoid elicited significantly greater serologic responses compared to controls for all antigens. The modified-live vaccine elicited a significantly greater response compared to controls for a whole-cell antigen preparation. Lesion scores were significantly smaller (greater protection) in calves that received the bacterin-toxoid, but not the modified-live vaccine, compared to controls.

Role of tissue factor in intra-alveolar fibrin deposition and coagulopathy associated with pneumonic pasteurellosis in cattle.

OBJECTIVE: To determine the role of tissue factor (TF) in the coagulation events leading to intra-alveolar fibrin deposition and intravascular thrombosis associated with pneumonic pasteurellosis in cattle. ANIMALS: Healthy 2- to 4-week-old male Holstein calves. PROCEDURES: Blood and bronchoalveolar lavage samples were collected before and at 1, 2, 4, and 6 hours after inoculation of saline solution or Pasteurella haemolytica. Total leukocyte count, platelet count, plasma total protein concentration, prothrombin time, and partial thromboplastin time were measured in blood samples. Total nucleated cell count, total protein concentration, and procoagulant activity were measured in bronchoalveolar lavage samples. Additionally, platelet survival in blood platelet accumulation in affected lung tissue, and gross and microscopic lung lesions were determined. RESULTS: Administration of TF monoclonal antibodies (MAB) TF1-1F7 prevented the decrease in platelet survival and the increase in bronchoalveolar lavage fluid TF-dependent procoagulant activity observed in calves not treated with MAB TF1-1F7 antibody, but did not attenuate the increase in lavage fluid neutrophil numbers and total protein concentration, MAB TF1-1F7 administration reduced the percentage of lung affected by pneumonic lesions from 51.81% to 10.40% and attenuated intra-alveolar deposition of fibrin, neutrophils, and erythrocytes. CONCLUSION: Intra-alveolar fibrin deposition and activation of coagulation in cattle with pneumonic pasteurellosis is, at least in part, mediated by TF. CLINICAL RELEVANCE: Treatments that neutralize TF activity may attenuate lung injury in cattle with pneumonic pasteurellosis.

[Experimental affecting of pulmonary clearance of Pasteurella multocida induced pneumonia in swine]. / Untersuchungen zur experimentellen Beeinflussung der Lungenclearance und der durch Pasteurella multocida induzierten Pneumonie beim Schwein.

The pulmonary clearance of Pasteurella multocida in weaner pigs was not affected by Carrageenan and Silica, two substances which block the function of monocytes/macrophages. Neutropenia, caused by the application of hydroxyurea, however, inhibit the pulmonary clearance markedly and reduced the severity of simultaneously induced pneumonias considerably. This indicates the importance of polymorphonuclear neutrophils for early clearance mechanisms and their inflammation inducing and maintaining effects.

Inducción de la reacción de Shwartzman en el pulmón del conejo mediante el empleo de lipopolisacárido de Pasteurella haemolytica / Induction of the Shwartzman reaction in the rabbit lung employing Pasteurella haemolytica lipopolysacharide

Se indujo la reacción de Shwartzman (RS) en el pulmón de conejo empleando lipopolisacárido (LPS) de Pasteurella haemolytica con el fin de comparar las lesiones provocadas con las que se presentan naturalmente en la pasteurelosis neumónica (PN). Para inducirla, primero se administró una dosis de LPS de Pasteurella haemolytica (50 mg) por vía endotraqueal (inoculación preparatoria) seguida 24 h más tarde por otra dosis del mismo LPS (100 mg) por vía endovenosa (inoculación desencadenante) (Grupo 5). Doce horas después, los animales fueron sacrificados. El pulmón derecho se destinó a estudios de histopatología, mientras que el izquierdo se empleó para realizar lavados bronquioalveolares (LBA). Para comparar se incluyeron grupos que recibieron solamente el LPS de P. haemolytica por vía endotraqueal (Grupo 3) o endovenosa (Grupo 4) (la inoculación faltante correspondió a solució salina fisiológica [SSF]), así como otro grupo al que se le administró LPS de Escherichia coli tal y como se describió al principio para inducir la RS (Grupo 2), y otro más que recibió SSF de la misma manera (Grupo 1). Las lesiones más notorias se presentaron en los animales que recibieron LPS de P. haemolytica por vía endotraqueal y en los que se les provocó la RS con el mismo LPS. Sólo los polimorfonucleares (PMN), monocitos y linfocitos recuperados por LBA resultaron diferentes entre los grupos, particularmente en los animales que recibieron el LPS de P. Haemolytica por vía endotraqueal y a los que se les indujo la RS con el mismo LPS. Se concluye que el LPS de P. Haemolytica es capaz de provocar una reacción flogística en el pulmón tanto por vía endotraqueal como endovenosa, aunque la respuesta es mucho más intensa en la primera, además, la RS no provoca una respuesta inflamatoria más intensa que la sola inoculación endotraqueal del LPS. Finalmente, se puede afirmar también que el conejo es un buen modelo para evaluar la respuesta inflamatoria pulmonar provocada por el LPS de P. haemolytica

The goat as a model for studies of pneumonic pasteurellosis caused by Pasteurella multocida.

A model of pneumonic pasteurellosis has been established in goats using Pasteurella multocida harvested from pneumonic lungs of goats (types A and D), rabbits (type A) and sheep (type D). The resultant infections were acute, subacute or chronic. The gross and histological lesions of the subacute and chronic infections were typical of pneumonic pasteurellosis. P. multocida type D produced significantly (P < 0.01) more severe lesions when compared with other isolates. There were strong correlations between the clinical signs and the severity of lesions.

Pulmonary ventilation, mechanics, gas exchange and haemodynamics in calves following intratracheal inoculation of Pasteurella haemolytica.

A Pasteurella haemolytica A1 broth was injected intratracheally in eight calves and measurements of pulmonary function values (PFV) were made once before and hourly post inoculation (p.i.). Changes in PFVs, included increased respiratory rate and minute ventilation (up to 158% of baseline 2 h p.i.) and decreased tidal volume and lung dynamic compliance (up to 33% of baseline 3 h p.i.). Total pulmonary resistance was not affected. At and after 3 h p.i. there was a progressive impairement of gas exchange, as judged from arterial O2 tension which decreased up to 65% of baseline. In contrast, arterial CO2 tension was not affected. Pulmonary hypertension was observed during the 3 last h of the study and was attributable to an increased pulmonary vascular resistance. Severe neutropenia was observed at 3 h p.i. and post-mortem histological findings were consistent with an acute fibrinohemorragic bronchopneumonia. In conclusion, P. haemolytica airway challenge unequiovocally resulted in acute pneumonia, providing a reproducible pathophysiological model for investigations regarding new therapeutic strategies.

Economic, clinical and functional consequences of a treatment using metrenperone during an outbreak of shipping fever in cattle.

Eighteen of 91 seven- to nine-month-old Belgian white and blue double-muscled male fattening cattle developed typical signs of shipping fever. They were all injected intramuscularly once a day for three days with 5 mg/kg of enrofloxacin, and in addition nine selected at random were injected intramuscularly five times at 12 hour intervals with 0.1 mg/kg of metrenperone, a 5-hydroxytryptamine blocker, the other nine receiving a placebo. During the outbreak of shipping fever metrenperone showed effective antipyretic properties, and all the calves treated with it made a complete recovery. Moreover, during the 360 day fattening period following the outbreak, the cattle treated with metrenperone gained on average 45.4 kg more weight than the control cattle.

Experimental reproduction of acute pneumonic pasteurellosis in rabbits.

A histomorphometric and physiopathological study was made of the lung parenchyma of Belgian White SPF rabbits infected experimentally with Pasteurella multocida type A. Symptoms observed were characteristic of the acute respiratory syndrome. Mean serum cortisol concentration and rectal temperature increased in all experimental groups. Histopathological changes included alveolitis and leukocytic bronchitis. Changes in alveolar and bronchial cytoarchitecture were attributed to the degeneration and necrosis of constituent epithelial cells.

Effect of enrofloxacin therapy on shipping fever pneumonia in feedlot cattle.

The effect of enrofloxacin therapy was investigated in 110 male double-muscled cattle weighing 275 +/- 3 kg, during a spontaneous outbreak of shipping fever occurring 11 +/- 2 days after they arrived in the feedlot. Forty-six diseased animals were divided randomly into three groups A, B and C, containing 17, 19 and 10 animals, respectively; the animals in group A were injected intramuscularly once daily for three consecutive days with 2.5 mg/kg of enrofloxacin, those in group B with 5 mg/kg of enrofloxacin and those in group C with 10 mg/kg of oxytetracycline. Clinical, serological, production and respiratory functional observations were recorded. The animals were clinically cured after the three day treatment except for three in group A and two in group C. These five animals made a clinical recovery after a three day booster treatment with a dose of 5 mg/kg enrofloxacin. The changes in respiratory gas exchange values induced by shipping fever were completely reversed 15 days later, suggesting that there had been no irreversible lung damage. The daily weight gains and the arterial blood gas values of the three groups of treated cattle were not significantly different. The high efficacy of the low dosage of enrofloxacin in this clinical syndrome may be explained by its antibacterial activity against Pasteurella species and Mycoplasma species. This field trial supports the in vitro studies which suggested than enrofloxacin is an appropriate therapy in cases of shipping fever.

The effect of common bovine respiratory diseases on tidal breathing flow-volume loops.

In order to better understand the bovine breathing pattern, tidal breathing flow-volume loops (TBFVL) were analyzed in 24 healthy cattle of different body weights (range: 37-660 kg) (Group A) and in 28 cattle suffering from the common respiratory diseases: verminous bronchitis (Group B); shipping fever (Group C); acute respiratory distress syndrome (Group D); respiratory syncytial virus pneumonia (Group E); organophosphate poisoning (Group F); and necrotic laryngitis (Group G). Respiratory airflow and tidal volume were measured with a breathing mask-Fleisch pneumotachograph assembly. TBFVL were traced from these values using a computerized method. All the loop indices proposed by Amis and Kurpershoek (1986a) were calculated from 5 representative breathing cycles for each of the 52 animals. The TBFVL shapes and indices were relatively constant in most healthy cattle and were not correlated with the body size. When compared to normal values, animals with moderate respiratory syndromes (Groups B and C) had a more flattened shape to their TBFVL. On the other hand, in most cattle with severe respiratory pathologies (Groups D, F and G expiration tended to be biphasic with the peak expiratory flow (PEF) occurring significantly later than in healthy animals. Both PEF and peak inspiratory flow were increased in all the pathological conditions. The TBFVL indices were more frequently and more severely changed during expiration than during inspiration.

Cellular inflammatory response in the lungs of calves exposed to bovine viral diarrhea virus, Mycoplasma bovis, and Pasteurella haemolytica.

Three, 5, or 7 days after inoculation with bovine viral diarrhea (BVD) virus (n = 12) or Mycoplasma bovis (n = 12), groups of calves were exposed to aerosols of Pasteurella haemolytica and were euthanatized 4 hours later. Histologic lesions in the lungs and the ratios of neutrophils to alveolar macrophages, collected by bronchoalveolar lavage, were compared with those of clinically healthy calves (n = 8) and calves inoculated with BVD virus only (n = 4), M bovis only (n = 4), or P haemolytica only (n = 2). Inoculation with BVD virus or M bovis did not have a significant (P greater than 0.05) effect on the neutrophil/macrophage ratio in the bronchoalveolar lavage. Aerosol exposure to P haemolytica induced a marked and significant (P less than 0.01) change in the neutrophil/macrophage ratio (from less than 1:9 to greater than 9:1). The reversed neutrophil/macrophage ratio in calves exposed to P haemolytica correlated well with the histologic changes in which small bronchi and bronchioles were plugged with purulent exudate. Inoculation with BVD virus did not induce gross or microscopic lesions in the lungs. Inoculation with M bovis resulted in a severe peribronchial lymphoid hyperplasia with mild exudation of neutrophils and macrophages into the cranioventral parts of the lungs.

Hemodynamic effects of acute pneumonia experimentally induced in newborn calves inoculated with Pasteurella haemolytica.

Hemodynamic responses to acute pneumonia and to hypoxia were investigated in 10 newborn calves. Experiments were performed on heparinized, anesthesized and ventilated calves. Control calves were inoculated intratracheally with bovine fetal serum. Pneumonia was induced in treated calves by intratracheal inoculation with P haemolytica suspended in bovine fetal serum. Before inoculation (base line), at the time of inoculation (T = 0), and at 30-minute intervals for 3 hours, pulmonary arterial pressure (Ppa), systemic arterial pressure, cardiac output (CO), arterial blood gases, pulmonary vascular resistance (PVR), and systemic vascular resistance were determined. At T = 0, calves in both groups became hypoxemic, alveolar-arterial O2 difference, PVR, and Ppa increased, and CO and systemic vascular resistance remained unchanged. At subsequent measurement intervals, all values returned to base-line in control calves, whereas treated calves had progressive hypoxemia associated with a decrease in Ppa and PVR, with no change in CO. Three hours after inoculation and after inhalation of 10% O2 in N2, PVR increased significantly in the control calves. In the treated group, hypoxia did not increase the resistance, compared with base-line and 3-hour values. The data indicate decreased Ppa during pneumonic pasteurellosis is because of a decrease in PVR and that pneumonia may attenuate the normal pulmonary hypoxic vasoconstrictor response.

The possible role of stress in the induction of pneumonic pasteurellosis.

Five groups of range bred calves (four calves per group) were used to investigate the effect of stress on susceptibility to aerosol exposures with bovine herpesvirus-1 or Pasteurella haemolytica. Twelve calves were weaned, transported, processed at a commercial feedlot and transported to isolation facilities three days later. An aerosol challenge of either 10 colony forming units of P. haemolytica or 10 plaque forming units of bovine herpesvirus-1 virus was given to two groups of calves and the third group was not challenged. The fourth group was transported directly to the isolation facilities after weaning and aerosol challenged with P. haemolytica. The fifth group remained at the farm after weaning and was not challenged. All transported animals had elevated plasma cortisol levels which remained above normal for at least three days postchallenge. The blastogenic response of all calves was depressed after leaving the farm and remained depressed throughout the experiment. The suppression correlated well with elevated serum cortisol levels. Calves processed through the feedlot encountered bovine herpesvirus-1 because eight out of 12 animals seroconverted to this antigen. Most calves seroconverted to P. haemolytica whether they were experimentally challenged or not. Where the unchallenged calves encountered P. haemolytica is unknown. Calves challenged with bovine herpesvirus-1 but not with P. haemolytica, had significant clinical signs of pneumonia and two animals died due to bovine herpesvirus-1 infection.(ABSTRACT TRUNCATED AT 250 WORDS)