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.     

Effect of a new macrolide antibiotic (tilmicosin) on pneumonia experimentally induced in calves by Mycoplasma bovis and Pasteurella haemolytica

Two gnotobiotic calves were treated once with tilmicosin (20 mg kg-1) six hours before they were infected by the intratracheal route with Mycoplasma bovis and Pasteurella haemolytica serotype 1. This treatment prevented colonisation of the lungs by P haemolytica and considerably reduced colonisation by M bovis, and the clinical scores and the extent of pneumonic consolidation, compared with two untreated gnotobiotic calves, both of which had to be killed in extremis for humanitarian reasons within 24 hours of infection. In a second experiment, 10 conventionally reared calves were similarly exposed to infection and, at the onset of clinical disease, five were treated once with tilmicosin (20 mg kg-1). Colonisation by P haemolytica and M bovis, the clinical scores and extent of pneumonic consolidation were suppressed or greatly reduced in the treated compared with the untreated calves, one of which had to be killed in extremis two days after infection. It was concluded that tilmicosin had a beneficial effect.     

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)     

Treatment of experimentally induced pneumonic pasteurellosis of young calves with tilmicosin.

Twenty four (24) healthy male Holstein calves (< 70 kg) were each experimentally infected by intrabronchial inoculation of 4.0 x 10(9) viable cells of Pasteurella haemolytica-AI (B122) at Time = 0 h. At 1 h following inoculation animals received either: 1) Sham treatment with sterile 0.85% saline SC (n = 12); or 2) a single injection of 10 mg tilmicosin per kg body weight (n = 12). Calves that were non-infected and tilmicosin-treated were also included for determining tilmicosin concentrations in serum and lung tissue at 1, 2, 4, 6, 8, 24, 48, and 72 h (n = 3-per time). In the infected calves, response to therapy was monitored clinically. Serum samples were collected for determination of tilmicosin concentrations using HPLC. Any animal becoming seriously ill was humanely killed. Complete necropsy examinations were performed on all animals and included gross pathologic changes, bacteriologic analysis, histopathology, and determination of pulmonary concentrations of tilmicosin. Tilmicosin treated animals responded significantly better to therapy than saline-treated control calves. Clinical assessment of calves during the study indicated that tilmicosin-treated calves had significantly improved by T = 8 h compared to satine-treated animals (P < 0.05). At necropsy tilmicosin-treated calves had significantly less severe gross and histological lesions (P < 0.05) of the pulmonary tissue. Of the 12 saline-treated calves, 92% (11/12) had Pasteurella haemolytica-A1 in lung tissue, while of the tilmicosin-treated calves 0% (0/12) cultured positive for P. haemolytica. Mean (+/- standard error) serum tilmicosin concentrations in infected calves peaked at 1 h post-injection (1.10 +/- 0.06 micrograms/mL) and rapidly decreased to 0.20 +/- 0.03 microgram/mL, well below the MIC of 0.50 microgram/mL for P. haemolytica-A1 (B122), by 12 h. These serum concentrations were very similar to serum concentrations of tilmicosin in non-infected tilmicosin-treated calves. Lung tissue concentrations of the antibiotic were comparatively high, even at 72 h post-infection (6.50 +/- 0.75 ppm). Lung tissue concentrations at 72 h were significantly higher in experimentally infected calves than in non-infected tilmicosin-treated animals (P < 0.05). These data demonstrate that tilmicosin was effective in treating experimentally-induced pneumonic pasteurellosis as determined by alleviation of clinical signs, pathological findings at post mortem, and presence of viable bacteria from the lung. Concentrations substantially above MIC for P. haemolytica were present in lung tissue even at 72 h following a single subcutaneous injection of 10 mg tilmicosin per kg body weight.     

Comparison of pharmacokinetics of danofloxacin and enrofloxacin in calves challenged with Mannheimia haemolytica

OBJECTIVE: To compare concentrations of danofloxacin, enrofloxacin, and ciprofloxacin in plasma and respiratory tissues of calves treated after challenge with Mannheimia haemolytica. ANIMALS: 75 calves. PROCEDURE: 24 hours after challenge with M. haemolytica, 72 calves with clinical signs of respiratory tract disease were randomly assigned to 1 of 12 equal treatment groups.Three nonchallenged, nontreated calves formed a control group. Challenged calves were treated with danofloxacin (6 and 8 mg/kg, SC) and enrofloxacin (8 mg/kg, SC) once. At 1, 2, 6, and 12 hours after treatment, 6 calves from each treatment group were euthanatized. Antimicrobial drug concentrations were assayed in various specimens. Peak plasma concentration (Cmax)-to-minimum inhibitory concentration (MIC; Cmax-to-MIC) ratios and the area under the concentration versus time curve over a 12-hour period-to-MIC ratios (AUC(12h)-to-MIC) were calculat-ed. RESULTS: Danofloxacin and enrofloxacin had MICs of 0.03 microg/mL for the M. haemolytica challenge isolate. Danofloxacin administered at doses of 6 and 8 mg/kg resulted in numerically higher geometric mean concentrations of danofloxacin in plasma and all respiratory tissues than geometric mean concentrations of enrofloxacin after treatment with enrofloxacin. Geometric mean concentrations of enrofloxacin were numerically higher than geometric mean concentrations of ciprofloxacin metabolite in plasma and almost all respiratory tissues. Danofloxacin and enrofloxacin achieved Cmax-to-MIC ratios >10 and AUC(12h)-to-MIC ratios >125 hours. CONCLUSIONS AND CLINICAL RELEVANCE: When used to treat pneumonic pasteurellosis in calves, danofloxacin and enrofloxacin can be expected to deliver concentration-dependent bactericidal activity against M. haemolytica, the bacteria most commonly associated with bovine respiratory tract disease.     

Efficacy of danofloxacin in the treatment of respiratory disease in European cattle

The efficacy of an injectable formulation of danofloxacin (180 mg/ml) in the treatment of naturally occurring bovine respiratory disease was evaluated in field studies on farms in France, Ireland and the United Kingdom. Cattle aged one week to 15 months with clinical respiratory disease were randomly allocated to treatment with 6 mg/kg danofloxacin or 10 mg/kg tilmicosin, administered by a single subcutaneous injection on day 0. A second injection of danofloxacin was administered on day 2, only if predefined clinical criteria were met. Mannheimia haemolytica, Pasteurella multocida and Haemophilus somnus were isolated from pretreatment nasopharyngeal swabs taken on all the farms. After the treatment, there was a more rapid improvement in the clinical response of the 178 animals treated with danofloxacin by day 2 (P < 0.01) than in the 90 treated with tilmicosin. For both treatments, there were similar significant (P < 0.001) reductions in the mean rectal temperature and severity of clinical signs of abnormal respiration and depression, on days 4 and 10 compared with day 0; 78.1 per cent of the animals treated with danofloxacin and 78.5 per cent of those treated with tilmicosin completed the studies. Danofloxacin 18 per cent was clinically safe and as effective as tilmicosin in the treatment of bovine respiratory disease.     

Clopidogrel induced suppression of bovine platelet activation in vitro and a preliminary study of its effect on the development of Mannheimia haemolytica induced pneumonia

We report here on the influence of the platelet antagonist clopidogrel (Plavix) on bovine platelet function. We first evaluated the capacity of clopidogrel to inhibit adenosine diphosphate (ADP)-stimulated platelet function in the bovine species, using an ex vivo approach with blood from treated animals. Platelets isolated from treated calves displayed rapid and consistent reduction in function (aggregation, thromboxane production) upon ADP, but not platelet activating factor (PAF), stimulation. We then examined the possibility that clopidogrel could influence Mannheimia haemolytica pneumonia pathobiology using an experimental challenge model. We were unable to detect significant differences between clopidogrel treated and untreated animals when challenged with intra-tracheal inoculation of M. haemolytica. There was a trend towards inhibition of platelet degranulation in the affected regions of lungs from clopidogrel treated calves, and pre-treated challenged animals had similar amounts of fibrin deposition and enhanced fibrous tissue formation in their lungs when compared with control counterparts.     

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.     

A comparison of florfenicol and tilmicosin for the treatment of undifferentiated fever in feedlot calves in western Canada

A field trial was performed under commercial feedlot conditions in western Canada to compare the efficacy of florfenicol and tilmicosin for the treatment of undifferentiated fever (UF) in calves that received metaphylactic tilmicosin upon arrival at the feedlot. One thousand and eighty recently weaned, auction market derived, crossbred beef calves suffering from UF were allocated to one of 2 experimental groups as follows: florfenicol, which was intramuscular (i.m.) florfenicol administered at the rate of 20 mg/kg body weight (BW) at the time of allocation (Day 0) and again 48 h later, or tilmicosin, which was subcutaneous (s.c.) tilmicosin administered once at the rate of 10 mg/kg BW on day 0. Five hundred and forty-four animals were allocated to the florfenicol group and 536 animals were allocated to the tilmicosin group. The chronicity, wastage, overall mortality, and bovine respiratory disease (BRD) mortality rates were significantly (P < 0.05) lower in the florfenicol group than in the tilmicosin group. There were no significant (P > or = 0.05) differences in first UF relapse, second UF relapse, hemophilosis mortality, or miscellaneous mortality rates between the florfenicol and tilmicosin groups. Average daily gain (ADG) from arrival at the feedlot to the time of implanting and ADG from allocation to the time of implanting were significantly (P < 0.05) lower in the florfenicol group as compared with the tilmicosin group. There were no significant (P > or = 0.05) differences in arrival weight, allocation weight, implanting weight, or ADG from arrival to allocation between the experimental groups. In the economic analysis, there was an advantage of $18.83 CDN per animal in the florfenicol group. The results of this study indicate that florfenicol is superior to tilmicosin for the treatment of UF because of lower chronicity, wastage, overall mortality, and BRD mortality rates. However, interpretation of these observations must take into consideration the fact that these calves received meta-phylactic tilmicosin upon arrival at the feedlot, which is a standard, cost-effective, management procedure utilized by feedlots in western Canada.     

Efficacy of difloxacin in calves experimentally infected with Mannheimia haemolytica

OBJECTIVE: To determine the efficacy of difloxacin, a novel fluoroquinolone antibiotic, in calves experimentally infected with Mannheimia haemolytica (formerly Pasteurella haemolytica). ANIMALS: Seventy-two 3-month-old Holstein calves. PROCEDURES: Calves were inoculated with M haemolytica intratracheally; after they developed clinical signs of pneumonic pasteurellosis, they were randomly assigned to 1 of 6 groups (n = 12/group). Calves in each group were treated with 10% difloxacin (2.5 or 5 mg/kg of body weight), 5% difloxacin (2.5 or 5 mg/kg), enrofloxacin (5 mg/kg), or saline (0.9% NaCl) solution (control group), once daily for 5 days, and clinical signs were scored daily. On day 15, calves were euthanatized, and the percentage of diseased lung tissue was calculated. Swab specimens of the lungs were submitted for bacterial culture. RESULTS: Mortality rate and percentage of diseased lung tissue were significantly higher and cure rate and average daily gain were significantly lower for control calves, compared with calves in the treatment groups; however, no significant differences were found among treatment groups. Mannheimia haemolytica was isolated from the lungs of 10 control calves and from at least 2 calves in each of the treatment groups. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that difloxacin and enrofloxacin were equally effective for treatment of calves with experimentally induced pneumonic pasteurellosis. However, treatment of infected calves with difloxacin or enrofloxacin may not eliminate the organism.     

Comparison of bronchoalveolar lavage fluid obtained from Mannheimia haemolytica-inoculated calves with and without prior treatment with the selectin inhibitor TBC1269

OBJECTIVES: To determine effects of selectin inhibitor TBC1269 on neutrophil infiltration, and neutrophil-associated injury during pneumonia induced by Mannheimia haemolytica and concentration of antimicrobial anionic peptide (AAP) in bronchoalveolar lavage fluid (BALF) as well as antimicrobial activity of BALF from healthy (control) neonatal calves, neonatal calves with M haemolytica-induced pneumonia, neonatal calves with prior treatment with TBC1269, and adult cattle. ANIMALS: Eighteen 1- to 3-day-old calves and 9 adult cattle. PROCEDURE: Calves were inoculated with M haemolytica or pyrogen-free saline (0.14M NaCl) solution into the right cranial lung lobe, and BALF was collected 2 or 6 hours after inoculation. Thirty minutes before and 2 hours after inoculation, 4 calves received TBC1269. The BALF collected from 9 adult cattle was used for comparison of BALF AAP concentration and antimicrobial activity. Protein concentration and neutrophil differential percentage and degeneration in BALF were determined. An ELISA and killing assay were used to determine BALF AAP concentration and antimicrobial activity, respectively. RESULTS: Total protein concentration was significantly decreased in BALF from calves receiving TBC1269. Similar concentrations of AAP were detected in BALF from all calves, which were 3-fold higher than those in BALF from adult cattle. However, BALF from neonates had little or no anti-M haemolytica activity. CONCLUSIONS AND CLINICAL RELEVANCE: These results suggest that TBC1269 decreases pulmonary tissue injury in neonatal calves infected with M haemolytica. Although AAP is detectable in neonatal BALF at 3 times the concentration detected in adult BALF, neonatal BALF lacks antimicrobial activity for M haemolytica.     

Effect of bovine granulocyte colony-stimulating factor on the development of pneumonia caused by Mannheimia haemolytica

The role of recruited neutrophils in Mannheimia haemolytica infection is controversial. We hypothesized that the neutrophilia induced by recombinant bovine granulocyte colony-stimulating factor (GCSF) would lead to rapid bacterial clearance and less severe lesions after infection with M. haemolytica. Two experiments (A and B) were conducted in which four calves per experiment were treated daily with 5 microg/kg GCSF and four calves per experiment were treated with saline. All 16 calves were challenged with 5 x 10(9) colony-forming units (cfu)/ml (experiment A) or 4.5 x 10(8) cfu/ml (experiment B) of M. haemolytica bacteria, into the right bronchus by bronchoscope-placed catheter. The mean maximal blood neutrophil counts in non-GCSF-treated and GCSF-treated calves before bacterial challenge were 5.6 +/- 0.7 x 10(9)/liter and 25.4 +/- 2.7 x 10(9)/liter, respectively. Two untreated calves became neutropenic and were euthanatized 2 days after infection because of severe respiratory distress. GCSF-treated calves had a 37% reduction in lung lesions compared with nontreated calves, and this difference was significant (P=0.04) when the effect of previous antibody titre to leukotoxin was considered. The effect of GCSF treatment on the severity of clinical signs seemed to be influenced by the antibody titre to M. haemolytica leukotoxin, although this effect could not be conclusively addressed. In conclusion, GCSF induced neutrophilia and partially protected calves against experimental infection with M. haemolytica. These results imply that increased numbers of neutrophils may, under some circumstances, protect against severe pneumonia caused by M. haemolytica.     

A field comparison of the efficacy and tolerance of marbofloxacin in the treatment of bovine respiratory disease

A multicentre, controlled, randomized and blinded trial was carried out in 180 ruminating calves with pyrexia and respiratory sign(s) on nine Belgian, British and French farms. All animals were sampled for pathogenic bacteria before treatment and at failure/relapse. Calves were injected with either marbofloxacin (M) solution [Marbocyl (Laboratoire Vétoquinol, Lure, France) 10%] at 2 mg/kg/24 h for 4 days intravenously on the first day then subcutaneously, or tilmicosin (T) solution (Micotil, Elanco Products Ltd, Basingstoke, Hants, UK) at 10 mg/kg as a single subcutaneous (s.c.) injection. The animals were examined clinically eight times up to day 28. The bacterial pathogens were found to be sensitive to marbofloxacin: for Pasteurella haemolytica the minimum inhibitory concentration (MIC)90 was 0.08 microg/mL and for P. multocida the MIC90 was 0.04 microg/mL. Cure rates at day 4 for group M and group T were 84 vs. 82%, respectively (P > or = 0.05). However, overall clinical score was significantly lower after 1 day in group M (P < 0.05). There was no difference in either relapse rate or average daily weight gain between groups. Marbofloxacin was found to be better tolerated than tilmicosin at the s.c. injection site (77.5 vs. 42.2% calves without local swelling, P=0.001) and was well tolerated when injected intravenously. Marbofloxacin was shown to have comparable but faster efficacy and better local tolerance than tilmicosin in the treatment of bovine respiratory disease (BRD).     

Tilmicosin does not inhibit interleukin-8 gene expression in the bovine lung experimentally infected with Mannheimia (Pasteurella) haemolytica.

The expression of the interleukin-8 (IL-8) gene was examined by in situ hybridization in lung tissues from calves experimentally infected with Mannheimia (Pasteurella) haemolytica and treated with tilmicosin. Interleukin-8 mRNA expression was detected in alveolar areas, particularly along interlobular septa, in the lumen, and in the epithelial cells of some bronchioles. In lesional lung tissues from animals that had received tilmicosin, we found large areas with limited inflammation. There was no staining for IL-8 mRNA in these areas. In contrast, in strongly inflamed areas, the same patterns and intensities of staining for IL-8 mRNA were detected in tilmicosin- and sham-treated animals. We conclude that tilmicosin does not affect the expression of IL-8 mRNA in tissue showing microscopic signs of inflammation. Together with previous reports, this supports the view that the pro-apoptotic properties of tilmicosin on neutrophils do not compromise the host defense mechanisms required to control the infection.     

The efficacy of danofloxacin in the therapy of pneumonia associated with Pasteurella species in housed calves

The efficacy of danofloxacin, a novel third generation fluoroquinolone, was assessed in the treatment of pneumonia in housed calves on three farms in the FDR and Italy. Seventy three calves with clinical signs of acute pneumonia and rectal temperatures greater than 40 degrees C were treated with danofloxacin at a dose rate of 1.25 mg/kg for three or five days depending on response to treatment. The response in these calves was compared to that obtained in 77 calves treated with trimethoprim/sulpha. The clinical response achieved with danofloxacin was superior to that achieved with trimethoprim/sulpha and significantly fewer calves which received danofloxacin required five days treatment. Pasteurella haemolytica and, or P. multocida were isolated from the majority of calves prior to treatment. All isolates were sensitive to danofloxacin and over 90 percent were sensitive to trimethoprim/sulpha.     

Cytokine expression by pulmonary leukocytes from calves challenged with wild-type and leukotoxin-deficient Mannheimia haemolytica

The objective of this study was to assess the role of leukotoxin (LKT) in modulating the pulmonary cytokine response of calves challenged with Mannheimia haemolytica. Thirty-six calves, seronegative to LKT and M. haemolytica whole cell antigen were divided into three groups (I, II and III). Calves in groups I and II were challenged by the intra-bronchial route with 25 mL of phosphate buffered saline (PBS) containing 0.44×10(9) cfu/mL of LKT deficient (lkt(-)) and 25 mL of PBS containing 0.31×10(9) cfu/mL of wild-type (wt) M. haemolytica serotype 1, respectively. Group III calves were challenged intra-bronchially with 25 mL of sterile PBS. Leukocytes were collected from broncho-alveolar lavage fluid (BALF) 4 days before and at 1, 3, and 6 days post-inoculation (p.i.). Expression of the following cytokines in the recovered leukocytes was measured using real-time PCR: interleukin (IL)-1β, -8, -10, -12 (p40) and TNF-α. The amount of TNF-α produced was also quantified by ELISA. Although a statistically significant difference in the expression of these cytokines was not observed between groups challenged with the wt and lkt(-) strains, the wt infected group (II) did exhibit higher mean clinical scores. Overall, there was considerable variation in the composition of the BALF between the groups and by day 7 p.i., both lkt(-)- and wt-challenged calves had seroconverted to M. haemolytica.     

The effect of parainfluenza virus type 3 on the phagocytic cell response of the ovine lung to Pasteurella haemolytica

Groups of Caesarean-derived, colostrum-deprived lambs were inoculated by the intratracheal route with Pasteurella haemolytica 4 to 6 days after the inoculation of parainfluenza virus type 3 (PI3). Some were killed immediately (0 h) and others 24 h later. Control groups were inoculated with PI3 alone, P. haemolytica alone or media alone. Pulmonary phagocytic cells, P. haemolytica and PI3 were recovered by pulmonary lavage. The phagocytes were separated into alveolar macrophage (AM) and neutrophil fractions by density gradient centrifugation and examined biochemically and microbiologically. Twenty-four hours after the inoculation of P. haemolytica bacterial proliferation to greater than 0 h levels had occurred in four of six animals inoculated with P. haemolytica alone, two of eight inoculated with P. haemolytica 4 days after PI3 and all of eight inoculated with P. haemolytica 6 days after PI3. Mean bacterial numbers in animals inoculated with P. haemolytica 6 days after PI3 and killed at 24 h (10(9.1 +/- 1.9)) were significantly higher than they were in the other two groups killed at this time (PI3 4 days, P. haemolytica 24 h, mean = 10(5.3 +/- 1.7); P. haemolytica alone 24 h, mean = 10(4.5 +/- 2.9)). Pneumonic lesions were also more severe in the first group. This defect in pulmonary clearance and increase in the severity of pneumonia in animals inoculated with P. haemolytica 6 days after PI3 coincided with a 1000-fold decrease in virus titres in the lung between Day 6 and Day 7 after virus inoculation and the first detectable evidence of the host's immune response. The virus infection resulted in a significant increase in the number of AM that could be recovered from the lung and an increase in the number of AM with cytoplasmic vacuolation. However, there was no difference in the total number of AM or the number of vacuolated AM between animals that controlled the P. haemolytica infection and those in which proliferation of P. haemolytica occurred. The inoculation of P. haemolytica resulted in a 100-fold increase in the number of neutrophils in the lavage fluid, but there were no differences between virus-infected and uninfected animals, nor was there a difference between animals that controlled the P. haemolytica infection and those that did not.(ABSTRACT TRUNCATED AT 400 WORDS)     

Effect of tilmicosin on chemotactic, phagocytic, and bactericidal activities of bovine and porcine alveolar macrophages

OBJECTIVE: To evaluate chemotactic, phagocytic, and bactericidal activities of bovine and porcine alveolar macrophages (AM) exposed to tilmicosin. ANIMALS: 12 healthy calves and 12 healthy pigs. PROCEDURES: Lungs were obtained immediately after euthanasia; AM were collected by means of bronchoalveolar lavage and density gradient centrifugation. Chemotactic activity was evaluated by exposing AM to lipopolysaccharide or macrophage inhibitory peptide during incubation with tilmicosin. Phagocytic activity was evaluated by incubating AM with tilmicosin for 24 hours and then with tilmicosin-resistant Salmonella serotype Typhimurium. Bactericidal activity was evaluated by incubating AM with tilmicosin (0, 10, or 20 microg/ml for bovine AM; 0 or 10 microg/ml or 10 microg/ml but washed free of tilmicosin for porcine AM) and then with Mannheimia haemolytica (bovine AM) or with Actinobacillus pleuropneumoniae or Pasteurella multocida (porcine AM). RESULTS: Tilmicosin had no significant effects on chemotactic or phagocytic activities of bovine or porcine AM. The time-course of bactericidal activity was best described by polynomial equations. Time to cessation of bacterial growth and area under the time versus bacterial number curve were significantly affected by incubation of AM with tilmicosin. CONCLUSIONS AND CLINICAL RELEVANCE: Results show that bactericidal activity of bovine and porcine AM was enhanced by tilmicosin, but not in proportion to the reported ability of AM to concentrate tilmicosin intracellularly. With or without exposure to tilmicosin, the time-course of bactericidal activity of bovine AM against M haemolytica and of porcine AM against A pleuropneumoniae or P multocida was too complex to be reduced to a simple linear equation.     

Immunogenicity of a soluble antigen against Pasteurella haemolytica-associated pneumonia in calves

Three experiments were performed to evaluate the immunogenic potency of a soluble fraction of Pasteurella haemolytica against pneumonic pasteurellosis in calves. A soluble antigen was extracted by a 2.5% saline solution from P. haemolytica. Weaned Holstein bull calves, seronegative for infectious bovine rhinotracheitis virus ( IBRV ) and the pasteurella antigen, were vaccinated either by repeated subcutaneous (SC) vaccination, or by exposure 3 times to the aerosol of P. haemolytica antigen. Challenge exposure to aerosol of P. haemolytica was preceded by infection with IBRV , or in experiments 2 and 3, the virus exposures were combined with a stress treatment. The lung lesions were examined at necropsy 3 to 8 days post infection. In the first experiment, all the vaccinated calves produced specific antibody response to the pasteurella antigen, and none of the calves including controls showed significant lesions in the lung. In the second experiment 2 aerogenically vaccinated calves had no lesions. One of the two SC-vaccinated calves had mild consolidated lesions. Two control calves, one of which died 3 days following the challenge, developed severe fibrinous pneumonia with consolidation of 50% or more of the lung surfaces. P. haemolytica was isolated only from the 2 control animals. In the third experiment, 2 of the 3 control calves developed moderate to severe consolidation, but P. haemolytica was isolated only from one of them. Two of the three aerosol-vaccinated calves also developed significant lesions and one of them yielded the bacteria from the lung. Three SC-vaccinated calves had slight lesions and the organism was not isolated from their lungs. The results did not consistently indicate an immunogenic potential of the soluble antigen against P. haemolytica-related pneumonia. The effect of stress on the pathogenesis of bovine viral pneumonia and correlation between pneumonic lesions and antibacterial resistance in situ are discussed.     

Induction of immunity against pneumonic pasteurellosis following experimental infection in calves.

Immunity against pneumonic pasteurellosis was studied in calves after recovery from experimental respiratory disease with Pasteurella haemolytica. Nine calves were exposed to aerosols of parainfluenza-3 virus and Pasteurella haemolytica A1 six days apart to produce respiratory disease. After recovery from the disease, these nine principal and four control calves were challenged with aerosols of bovine herpesvirus 1 and P. haemolytica A1 four days apart. With this viral-bacterial challenge, the nine principal animals failed to develop clinical responses to this bacterial challenge and their lungs did not show the growth of P. haemolytica on cultures, whereas two of four control calves had elevated temperatures and developed necropurulent pneumonia with the isolation of P. haemolytica from the lungs. The principal calves had developed high levels of cytotoxin neutralizing antibodies in their sera following parainfluenza-3 virus-P. haemolytica infection. This demonstrated that immunity against pneumonic pasteurellosis can be achieved, with a suggestion that further search for an effective vaccine for P. haemolytica is warranted.