Mannheimia Haemolytica A1 Research Articles

A Preliminary Investigation on a Commercial Ovine Pasteurellosis Vaccine Using Clinical and Pathological Endpoints.

IntroductionIn this study we aimed to provide preliminary evidence on the safety and efficacy of the currently used ovine pasteurellosis vaccine in Ethiopia using clinical and pathological endpoints.MethodsTwenty, conventionally reared, apparently healthy, seronegative male lambs, were randomly classified into two groups of 10 animals as “vaccinated-challenged” and ”unvaccinated-challenged controls”. The first group received 1 mL of the licensed Pasteurella multocida biotype A based vaccine subcutaneously while the second group received phosphate-buffered saline as a placebo. Following vaccination, lambs were monitored for one month for potential vaccine adverse reactions. Five weeks postvaccination, all lambs were immunosuppressed using dexamethasone, and intratracheally challenged with 5.2×109 CFU/mL live Mannheimia haemolytica A1 (clinical isolates). Then, all lambs were followed up for eight days for clinical examination and necropsied on the ninth day postchallenge for pathological investigation.ResultsThere were no safety issues recorded during the study. In terms of clinical signs, lambs developed fever, depression, mucoid bilateral oculonasal discharge, coughing and sneezing regardless of their vaccination status. Fisher’s exact test between vaccination status and each clinical sign showed a statistically insignificant association (p>0.05). The main pathological findings in both groups were pulmonary congestion, atelectasis, emphysema, and suppurative bronchopneumonia. Consolidation lung lesion score of +1 (5/10 of vaccinated, 6/10 of unvaccinated) and +2 (3/10 of vaccinated, 4/10 of unvaccinated) were recorded in a statistically indifferent manner among both vaccinated and nonvaccinated groups (p>0.05).Discussion and ConclusionCollectively, the results suggested that the vaccine posed no safety concern and presumably lacks protective efficacy against local isolates. However, the study did not analyze antibody titer and their functionality using serum bactericidal assays. Further confirmatory studies could provide more evidence on the vaccine efficacy. Safety should further be assessed in a field setting involving a large number of animals to enable detection of rare vaccine adverse events.

PSXVI-28 Late-Breaking: Effects of Preconditioning (Value Added Programs) on the Health, Performance, Mannheimia haemolytica, and Pasteurella multocida in Cattle Received on Winter Wheat Pasture

Abstract Bovine respiratory disease (BRD) is a persistent health problem impacting the beef industry. Research shows improved health and performance in preconditioned (PRECON) calves compared with nonpreconditioned (NONPRE) or commingled (COMM) calves received in the feedlot but little research has been focused on calves received on winter wheat pasture prior to feedlot entry. Our objective for this presentation is to investigate the effects of preconditioning on the health and performance of newly received beef calves on winter wheat pasture. Mixed breed steers (n = 145) were purchased from an auction barn in Dalhart, Texas, as PRECON (n = 70) or NONPRE (n = 75) and were transported to the Clayton Livestock Research Center in Clayton, New Mexico, for this 112-d study trial. Three treatments were used in this completely randomized design: PRECON (n = 50), NONPRE (n = 50) and COMM (n = 45). Upon arrival, steers were offloaded into separate pens. On d 0, steers were processed using a standard health protocol along with collection of nasopharyngeal (NP) swabs, randomly allocated to treatment, and released onto a 120-acre winter wheat pasture split into three paddocks with a common water source; weights were collected again on d 2, 90, and 112. There were no statistical differences in morbidity and mortality rates between treatments. Weight gain was analyzed using PROC GLM of SAS from d 0 to d 90. COMM steers had greater weight gains than PRECON (P = 0.04) and NONPRE (P = 0.02) steers. NP swabs were used to show the distribution of Mannheimia haemolytica (MH) serotype A1, A2, and A6 and Pasteurella multocida (PM) by day and by treatment. No statistical differences were observed in serotype distribution of MH A1, A2, or A6 or in PM. PRECON steers displayed no health or performance advantage over NONPRE or COMM steers.

Two outer membrane proteins are bovine lactoferrin-binding proteins in Mannheimia haemolytica A1.

Mannheimia haemolytica is a Gram negative bacterium that is part of the bovine respiratory disease, which causes important economic losses in the livestock industry. In the present work, the interaction between M. haemolytica A1 and bovine lactoferrin (BLf) was studied. This iron-chelating glycoprotein is part of the mammalian innate-immune system and is present in milk and mucosal secretions; Lf is also contained in neutrophils secondary granules, which release this glycoprotein at infection sites. It was evidenced that M. haemolytica was not able to use iron-charged BLf (BholoLf) as a sole iron source; nevertheless, iron-lacked BLf (BapoLf) showed a bactericidal effect against M. haemolytica with MIC of 4.88 ± 1.88 and 7.31 ± 1.62 μM for M. haemolytica strain F (field isolate) and M. haemolytica strain R (reference strain), respectively. Through overlay assays and 2-D electrophoresis, two OMP of 32.9 and 34.2 kDa with estimated IP of 8.18 and 9.35, respectively, were observed to bind both BapoLf and BholoLf; these OMP were identified by Maldi-Tof as OmpA (heat-modifiable OMP) and a membrane protein (porin). These M. haemolytica BLf binding proteins could be interacting in vivo with both forms of BLf depending on the iron state of the bovine.Electronic supplementary materialThe online version of this article (doi:10.1186/s13567-016-0378-1) contains supplementary material, which is available to authorized users.

Impact of Timing and Dosage of a Fluoroquinolone Treatment on the Microbiological, Pathological, and Clinical Outcomes of Calves Challenged with Mannheimia haemolytica.

The efficacy of an early and low inoculum-adjusted marbofloxacin treatment was evaluated on microbiological and clinical outcomes in calves infected with 4.107 CFU of Mannheimia haemolytica A1. Twenty-two calves were included based on their rectal temperature rise in the 10 h after challenge and allocated in four groups, receiving a single intramuscular injection of saline (CON), 2 mg/kg marbofloxacin 2–4 h after inclusion (early treatment, E2), 2 or 10 mg/kg marbofloxacin 35–39 h after inclusion (late treatments, L2, L10). In CON calves, M. haemolytica DNA loads in bronchoalveolar lavages continuously increased from inclusion to day 4, and were associated with persistent respiratory clinical signs and lung lesions. At times of early and late treatments, M. haemolytica loads ranged within 3.5–4 and 5.5–6 log10 DNA copies/mL, respectively. Early 2 mg/kg marbofloxacin treatment led to rapid and total elimination of bacteria in all calves. The late treatments induced a reduction of bacterial loads, but 3 of 6 L2 and 1 of 6 L10 calves were still positive for M. haemolytica at day 4. Except for CON calves, all animals exhibited clinical improvement within 24 h after treatment. However, early 2 mg/kg treatment was more efficacious to prevent pulmonary lesions, as indicated by the reduction of the extension and severity of gross lesions and by the histopathological scores. These results demonstrated for the first time that a reduced antibiotic regimen given at an early stage of the disease and targeting a low bacterial load could be efficacious in a natural bovine model of pneumonia.

An electrophoretic characterization of iron-transporting proteins in Mannheimia haemolytica A1

Iron-regulated outer membrane proteins (IROMPs) in Mannheimia haemolytica A1, which function as a receptor for complexes containing iron ions, are induced by iron deficiency in the growth environment of the bacteria. Densitometric analysis of SDS-PAGE separation showed expression of IROMPs of 71, 77, and 100 kDa in the case of bacteria grown in a medium with 2,2-dipyridyl. The electrophoregrams obtained in 2-DE separations confirmed the presence of protein fractions with these molecular weights and isoelectric points ranging from 5.4 to 6.4. The results of the study also confirmed the ability of M. haemolytica A1 proteins involved in iron uptake to induce a protective immune response. In Western blot with serum from convalescent calves naturally infected with M. haemolytica A1, distinct reactions were obtained for IROMPs of 71, 77, and 100 kDa.

In vivo gene expression in Mannheimia haemolytica A1 during a time-course trial in the bovine host

The objective of this study is to examine the expression of Mannheimia haemolytica genes over time during the early stage of infection. In addition, gene expression at different sites of infection in the bovine host was examined. A time-course experiment was designed to collect pharyngeal swabs and lung washings from the same animals over two time points. Six calves were experimentally challenged with M. haemolytica A1; pharyngeal swabs were collected from all animals 5h post infection. Three calves were euthanized at 6h; pharyngeal swabs were collected from the remaining 3 calves at 12h and the calves were euthanized. Lung washings were recovered from all animals at necropsy. Total RNA was prepared from the pharyngeal swabs and lung washings and primers for eight well characterized virulence-associated genes were used in qRT-PCR to examine mRNA levels. The expression of key virulence genes such as lktA, gcp and tbpB was higher in vivo compared to in vitro with the highest changes observed from 6 to 12h. The expression of lktA and gapA increased while expression of fbpA, gs60, nmaA and tbpB was found to decreased over time in the 6h period. Gene expression profiles in the lungs versus the pharynx also differed, with most genes (fbpA, tbpB, nmaA, gs60, lktA and narP) showing higher expressing in lung washings. This is the first study to follow gene expression by M. haemolytica in the same animal over time during an infection.

A proteomic analysis of the regulon of the NarP two-component regulatory system response regulator in the bovine pathogen Mannheimia haemolytica A1

BackgroundThe response of the NarQP two-component signal transduction system regulon in response to the presence of nitrate for the bovine pathogen Mannheimia haemolytica A1 was investigated by proteomic analysis. Total proteins from a narP mutant and the parent SH1217 grown with or without NaNO3 supplement were examined by ISO-DALT 2D electrophoresis and liquid chromatography-mass spectrometry.ResultsSeventeen proteins were differentially expressed in the parent strain SH1217 in response to the addition of NaNO3 to the growth media. These responses were absent in the narP mutant, indicating that the altered production of these proteins is mediated by NarPMh. Interestingly, NarPMh mediated the increased production of some proteins which are not generally associated with nitrate respiration, such as the iron transporters FbpA and YfeA. The increased production of proteins such as superoxide dismutase, SodA, and GAPDH were also observed. The increased production of these iron-regulated proteins by NarPMh is thought to enhance the swift establishment of the nitrate respiration mechanism of M. haemolytica during pathogenesis.ConclusionThe data suggested NarPMh acts as an important regulator which regulates the expression of a small set of proteins in response to nitrate availability. This may contribute to the prevalence of M. haemolytica A1 in its host during pathogenesis of BPP, through enhancing the effectiveness of nitrate respiration either directly or indirectly.

Sequence analysis of serotype-specific synthesis regions II of Haemophilus influenzae serotypes c and d: evidence for common ancestry of capsule synthesis in Pasteurellaceae and Neisseria meningitidis

Sequencing of yet unknown Haemophilus influenzae serotype c (Hic) and d (Hid) capsule synthesis regions II revealed four ( ccs1–4) and five ( dcs1–5) open reading frames, respectively. The inferred gene functions were in line with capsular polysaccharide structures. One or more proteins encoded by the Hic capsule synthesis region II showed similarity to Actinobacillus pleuropneumoniae serotype 1 and Actinobacillus suis K1 enzymes. Orthologues to the complete operon were observed in Actinobacillus minor strain 202, where even the gene order was conserved. Furthermore, Ccs4 was related to the capsule O-acetyltransferase of Neisseria meningitidis serogroup W-135. For the Hid locus, similarities to Hie, Mannheimia haemolytica A1 and N. meningitidis serogroup A were identified and the succession of genes was similar in the different species. The resemblance of genes and gene organization found for Hic and Hid with other species suggested horizontal gene transfer during capsule evolution across the bacterial classes.

Identification of putative two-component regulatory systems in the bovine pathogen Mannheimia haemolytica A1, and preliminary characterization of the NarQ/P system

The complete genome sequence of the bovine pathogen Mannheimia haemolytica A1 was analyzed by blast searches for the presence of two-component regulatory system proteins. Five complete sets of putative two-component systems were identified, and the NarQ/P system was further investigated. in silico analysis of the NarQ and NarP proteins showed features that are typical of the sensor and response regulator proteins. A narP knock-out mutant was constructed. The narP mutant has lost its ability to respond to NaNO(3) in the media and fail to alter the expression of several proteins. One of the proteins that showed increased production in the parent strain in response to NaNO(3) was analyzed by matrix-assisted laser desorption ionization time-of-flight MS. Unexpectedly, the protein was identified to be FbpA, a periplasmic component of the iron transporter system. Sequence analysis of the promoter region of fbpA identified motifs typical for NarP-regulated genes. The expression of the leukotoxin gene was also altered in the narP mutant as shown by Western immunoblot analysis and reverse transcription-PCR.

Electronic nose responses and acute phase proteins correlate in blood using a bovine model of respiratory infection

This study aimed (i) to assess the ability of electronic nose (e-nose) technology to differentiate between blood samples of experimentally infected and non-infected subjects and (ii) to evaluate e-nose responses given by volatile organic compounds in relation to the acute phase reaction generated in the host. In an animal model of gram-negative bacterial infection (20 calves; intratracheal inoculation of Mannheimia haemolytica A1), the concentrations of the acute phase proteins (APPs; i.e. lipopolysaccharide binding protein and haptoglobin) were measured in serum samples before and after challenge, and headspaces of pre- and post-inoculation serum samples were analysed using a conducting polymer-based e-nose. Significant changes of certain e-nose sensor responses allowed discrimination between samples before and after challenge. The maximal changes in responses of sensitive e-nose sensors corresponded to the peak of clinical signs. Significant correlations linked decreasing responses of multiple e-nose sensors to increasing concentrations of APPs in the peripheral blood.

Analysis of a collagen-binding trimeric autotransporter adhesin fromMannheimia haemolyticaA1

A locus that codes for a high-molecular-weight adhesin was previously isolated from Mannheimia haemolytica A1. In this study, we showed that this locus, named ahs, codes for two proteins (AhsA and AhsB) that exhibit characteristics of a trimeric autotransporter adhesin. Sequence analysis of AhsA showed the presence of 21 collagen-binding motifs in the protein. Collagen-binding assays showed that M. haemolytica A1 binds to collagen in a dose-dependent manner. This binding activity is trypsin sensitive and can be inhibited by anti-AhsA antibody. AhsB is the cognate transporter for AhsA. The C-terminal of AhsB showed highly conserved amino acids typical of trimeric autotransporters. Experimental data showed that the C-terminal 120 amino acids of AhsB could indeed form trimeric molecules. Western immunoblots showed the presence of anti-AhsA antibodies in the sera of calves that had been challenged with M. haemolytica A1, suggesting that AhsA is expressed and immunogenic in cattle.

Expression of a modified Mannheimia haemolytica GS60 outer membrane lipoprotein in transgenic alfalfa for the development of an edible vaccine against bovine pneumonic pasteurellosis

The GS60 antigen is one of the protective antigens of Mannheimia haemolytica A1. GS60 contains conserved domains belonging to the LppC family of bacterial outer membrane lipoproteins. A high antibody titer to GS60 has been shown to be significantly correlated with resistance to pneumonic pasteurellosis. Calves vaccinated with a commercial vaccine (Presponse™) and demonstrating protection against M. haemolytica A1 produced antibodies directed against GS60. Alfalfa was chosen as the platform for an edible vaccine. Agrobacterium tumefaciens was used to mediate the transformation of alfalfa with sequences encoding a slightly shortened derivative of the GS60 antigen (GS60 54). Stable transgenic alfalfa lines were recovered and production of GS60 54 was examined by Western immunoblot analysis. The antigen is stable in dried transgenic plant material stored at ambient temperature for more than a year. The plant-produced GS60 54 protein was shown to be immunogenic when injected into rabbits. Feeding of the dried transgenic alfalfa expressing the GS60 54 to rabbits is capable of inducing seroconversion, suggesting that GS60 54 could be an effective oral antigen for stimulating mucosal immune responses.

The Outer membrane protein OmpA ofMannheimia haemolyticaA1 is involved in the binding of fibronectin

An enzyme-linked immunosorbent assay using bovine fibronectin as the substrate was used to demonstrate that Mannheimia haemolytica A1 binds to fibronectin. This binding to fibronectin was specific as no binding was observed with bovine fibrinogen. The binding to fibronectin was not observed if the M. haemolytica A1 cells were pretreated with trypsin or proteinase K, suggesting that it involved a protein molecule on the cell surface. Interestingly, the fibronectin-binding activity was found to be higher in an acapsular mutant compared with its parent strain. The fibronectin-binding protein was shown to be present in the outer membrane fraction of M. haemolytica A1. A 45 kDa outer membrane protein that binds to fibronectin was identified by Far-Western immunoblot analysis. This protein was purified and subjected to MS matrix-assisted laser desorption ionization time-of-flight analysis. The results identified it to be outer membrane OmpA based on comparison with the M. haemolytica A1 genomic sequence.

Analysis of in vivo expressed genes in Mannheimia haemolytica A1

The expression of Mannheimia haemolytica A1 genes during in vivo growth was examined by reverse transcriptase-polymerase chain reaction (RT-PCR) using total RNA extracted directly from M. haemolytica A1 recovered from pneumonic lungs of cattle. Primers specific for three groups of genes were used. Group 1 includes virulence-related genes: lktC, tbpB, ahs, nmaA, gs60 and gcp. Group 2 includes genes that code for putative two-component regulatory systems: narP, narQ, ttrR, ttrS, phoB and phoR. Group 3 includes genes involved in regular cellular functions such as plp4, thiL and rrf. The RT-PCR data were examined in conjunction with the percent pneumonic lesion in each lung scored during necropsy. The analysis showed that lungs with a higher percent pneumonic score exhibit expression of more M. haemolytica A1 genes. For group 1 genes, lktC was expressed in the majority of samples, whereas the other genes were only expressed in some samples. This was not unexpected as the leukotoxin is a major virulence factor of the bacterium. The genes encoding the response regulators for the putative two-component regulatory systems were found to be expressed in more samples than the genes encoding the sensor proteins. The regulator proteins may be required in higher levels to regulate expression of target genes.

Cloning of a gene fragment encoding bovine complement component C3d with expression and characterization of derived fusion proteins

The gene fragment coding for bovine C3d gene ( boC3d) was cloned and expressed as a component of fusion proteins destined for use in vaccine studies in cattle, and for in vitro experiments. This fragment of complement protein C3 (C3d) has been shown to enhance B cell responses when complexed with antigen. Three potential vaccine constructs were engineered to contain one, two or three boC3d units linked to a fragment of the leukotoxin of Mannheimia haemolytica A1, an economically important pathogen of cattle that causes a fibrinous pneumonia in calves. A recombinant biotinylated boC3d protein (for use in in vitro studies) was generated by endogenous biotinylation in Escherichia coli by means of the BirA holoenzyme synthetase. All recombinant proteins incorporated polyhistidine tags and were purified by nickel-agarose chromatography, then analyzed by SDS-PAGE and Western immunoblot. The identity of boC3d was confirmed by mass spectrometry, since monoclonal antibodies to boC3d were not available. To date, published research into the adjuvant activities of C3d has been limited to experiments in mice and rabbits, using antigens unrelated to diseases occurring naturally in these species. The boC3d fusion proteins expressed in this study will provide the basis for immunization trials in cattle and studies of receptor binding and cell activation of bovine lymphocytes.

Challenge exposure of sheep immunized with live vaccine and culture supernatant of Mannheimia haemolytica A1: Effects of revaccination

Vaccination against Mannheimia haemolytica is a routine procedure in sheep farming, being repeatedly performed on the recommendations of commercial veterinary manufacturers or due to outbreaks of respiratory disease. However, the consequences of repeat vaccination after a short period, using live vaccines or culture supernatant (CS), have not been established, and its benefits are unknown. This study aims to evaluate, in a challenge exposure, the effects of revaccinating sheep with this type of biological product. Thus, 20 mixed-breed, 6–9-month-old sheep were divided into four groups, with five animals in each. Animals in group I were inoculated subcutaneously with 2 ml of a 1 × 10 9 CFU/ml suspension of bacteria in the exponential growth phase. Group II was inoculated with the same bacterial concentration but in the stationary phase. Animals in group III were inoculated with 3 ml of CS and group IV with saline solution, as control. Each group was immunized again 14 days later, following the same guidelines. On day 21, they were intranasally exposed to parainfluenza-3 virus (PI-3). On day 27, they were challenged with a transthoracic injection of live M. haemolytica (2 × 10 9 CFU/ml). In this experiment, the levels of agglutinating and antileukotoxin antibodies were evaluated through indirect hemagglutination and toxin neutralization assays. During the first 24 h post-challenge, four animals in group I died, three in group II and two in group III, even though they had developed significant levels of agglutinating and antileukotoxin antibodies. Only one animal from the control group died. Thus, contrary to the development of improved protection due to continuous immunizations with live bacteria or CS, a negative effect was observed. This can be explained as the result of a hypersensitivity type III reaction, due to the significant decrease in agglutinating antibodies observed shortly after the challenge, indicating the formation of immune complex, which triggered the release of chemical mediators of inflammation that, finally, promoted edema and pulmonary congestion.

Construction and analysis of a Mannheimia haemolytica A1 luxS mutant

Mannheimia haemolytica A1 is the causative agent of bovine pneumonic pasteurellosis, a major cause of sickness, death, and economic loss to the feedlot cattle industry. M. haemolytica A1 produces autoinducer-2 (AI-2) like molecules that are capable of inducing quorum sensing system 2 of Vibrio harveyi. This interspecies quorum sensing system has been shown to regulate the expression of virulence genes in several pathogenic bacteria. The protein central to the production of AI-2 is LuxS. To determine if quorum sensing is involved in the regulation of virulence genes in M. haemolytica A1, a luxS mutant was constructed by replacing luxS with a cat cassette. This mutant was verified by PCR analysis, Southern hybridization, as well as its inability to induce bioluminescence in the V. harveyi reporter strain. RT-PCR analysis showed there was no difference in leukotoxin (lktC) mRNA levels, however there were increased mRNA levels of putative virulence associated genes, transferrin binding protein B (tbpB), adhesin (ahs) and capsule biosynthesis (nmaA). Electron microscopy showed that the level of encapsulation in the mutant is higher than the parent. Additionally, the mutant was slightly more adherent to bovine tracheal cells than the parent. In vitro competition assays showed the mutant out-competed the parent under iron-restricted conditions. However, in a calf challenge, the parent was the dominant isolate recovered.

Incubation of bovine PMNs with conditioned medium from BHV-1 infected peripheral blood mononuclear cells increases their susceptibility to Mannheimia haemolytica leukotoxin

Active infection with bovine herpesvirus-1 (BHV-1) increases the susceptibility of cattle to secondary bacterial pneumonia with Mannheimia (Pasteurella) haemolytica A1. In the present study we found that bovine PMNs incubated with conditioned media from BHV-1 infected peripheral blood mononuclear cells (PBMCs) exhibited increased LFA-1 expression, enhanced LKT binding and increased LKT cytotoxicity. These effects were abrogated when the conditioned medium was pre-incubated with an anti-IL-1beta Mab before being added to the PMNs. These findings suggest that BHV-1 infection, and the resulting release of IL-1beta and perhaps other inflammatory cytokines, can stimulate activation of LFA-1 in bystander bovine PMNs, thus enhancing the binding and biological effects of LKT.

BHV-1 infection and inflammatory cytokines amplify the interaction of Mannheimia haemolytica leukotoxin with bovine peripheral blood mononuclear cells in vitro.

Bovine herpesvirus-1 (BHV-1) has been reported to increase the susceptibility of cattle to respiratory disease caused by Mannheimia (Pasteurella) haemolytica A1. The principal virulence factor of M. haemolytica is a leukotoxin (LKT) that can specifically kill ruminant leukocytes following its binding to the beta2-integrin CD11a/CD18 (lymphocyte function-associated antigen 1 (LFA-1)). In this study, we investigated the effects of experimental infection of bovine peripheral blood mononuclear cells (MNCs) with BHV-1 in vitro, on the subsequent interaction of these cells with the M. haemolytica LKT. We found that BHV-1 infection increased LFA-1 expression (as assessed by flow cytometry), and subsequently enhanced LKT binding and cytotoxicity to bovine MNCs. We also found that BHV-1 infection increased CD18, IL-1beta, and IFN-gamma mRNA expression by MNCs. As previously reported for bovine polymorphonuclear neutrophils (PMNs), MNCs increased their expression of LFA-1, and their LKT binding and cytotoxicity, following exposure to IL-1beta, TNF-alpha, and IFN-gamma. These findings suggest that BHV-1 infection, and the resulting release of inflammatory cytokines, can stimulate expression of LFA-1 in bovine MNCs, thus enhancing the binding and biological effects of LKT. If such a mechanism occurs in vivo it might explain, in part, the increased susceptibility of BHV-1 infected cattle to bovine pasteurellosis.

Association of LPS chemotype of Mannheimia (Pasteurella) haemolytica A1 with disease virulence in a model of ovine pneumonic pasteurellosis.

Host responses during pneumonic pasteurellosis were compared in sheep infected with strains of Mannheimia (Pasteurella) haemolytica A1 differing in their O-antigen type. Nine-week-old, specific pathogen-free lambs were infected intratracheally with parainfluenza type 3 virus (10(8) TCID(50)) followed 7 days later by 5-6 x 10(7) CFU of M. haemolytica A1 possessing rough (group R, 6 lambs) or smooth (group S, 6 lambs) lipopolysaccharide, or saline (group C, 4 lambs). Group C lambs remained afebrile with no evidence of endotoxaemia or bacteraemia and biochemical parameters were normal. Group R and group S lambs became febrile within 2-3 h postinfection and the response was higher and more prolonged in group R lambs. Four group R and 2 group S lambs developed clinical pneumonic pasteurellosis within 24-48 h and the severity of disease correlated with episodes of endotoxaemia, bacteraemia and elevated eicosanoid concentrations. At post-mortem, M. haemolytica (10(7)-10(9) CFU/g) was isolated from the lungs of all 6 group R lambs but from only 1 group S lamb. The results indicate an association between the incidence and severity of ovine pneumonic pasteurellosis and LPS chemotype and suggest an important role for LPS chemotype in determining host-species susceptibility to lung infection.