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1.
The outer membrane protein (OMP), plasmid, and antimicrobial resistance profiles of Pasteurella haemolytica serotypes 1 through 12, a bovine isolate of P multocida, a chicken isolate of P multocida, and an unidentified Pasteurella species of bovine origin were examined. Isolates of P haemolytica serotypes belonging to the same biotype possessed similar OMP profiles. Biotype A isolates contained 2 prominent OMP of 43 kilodaltons (kD) and 29 kD, whereas biotype-T serotypes contained 3 major OMP of 43, 36, and 25 kD. The major OMP profiles of the 2 P multocida isolates and the unidentified Pasteurella species were different from each other and from P haemolytica isolates. Plasmid DNA screening indicated both plasmid-containing and plasmid-free P haemolytica and P multocida isolates. Multiple drug resistance was found in pasteurellae isolates with and without plasmids. However, a relationship between drug resistance and plasmid isolation was found in 3 of 4 haemolytica serotype 1 field isolates, all of which contained a 2.51-megadalton plasmid and had multiple drug resistance for benzylpenicillin, ampicillin, streptomycin, and tetracycline.  相似文献   

2.
The biochemical and serological properties of 21 strains of Actinobacillus pleuropneumoniae biotype 2 isolated from haemorrhagic necrotic pleuropneumonia of swine were examined. For serologic typing, the indirect haemagglutination (IHA) and the double gel-diffusion tests were used. On the basis of their soluble surface antigens, our A. pleuropneumoniae biotype 2 isolates could be assigned to two proposed serotypes. Serotype 1 comprised 11 strains and serotype 2 comprised 10 strains. All strains contained two surface antigen components. In the strains belonging to serotype 1, one of the antigens was identical with the serotype-specific antigen of Pasteurella haemolytica T4. Both antigens of serotype 2 strains proved to be type-specific. Four strains received from Switzerland, including the holotype strain of A. pleuropneumoniae biotype 2, and three strains isolated from swine in the G.D.R. belonged to serotype 2. Both the double gel diffusion and the IHA tests detected a 2-way cross-reaction between biotype 1, serotype 2 and biotype 2, serotype 2 strains of A. pleuropneumoniae, which could be eliminated using cross-absorbed sera.  相似文献   

3.
Pasteurella haemolytica biotype A, serotype 1 (P haemolytica A1) was the most commonly isolated Pasteurella species from 80 calves examined at necropsy from 40 outbreaks of respiratory disease, the majority of which were pathologically confirmed as bovine pneumonic pasteurellosis (transit fever; shipping fever). Similarly, nasopharyngeal swabs from in-contact and apparently healthy calves indicated the widespread presence of P haemolytica A1. Pasteurella multocida and other serotypes of P haemolytica A1 were found including six isolations of P haemolytica T10, a fairly common pathogen in sheep. Approximately two-thirds of the isolates were tested for their antimicrobial sensitivity patterns and the degree of sensitivity for P haemolytica A1, the most frequently isolated serotype, was chloramphenicol (100 per cent), sulphamethoxazole trimethoprim (98 per cent), oxytetracycline (80 per cent), ampicillin (85 per cent), penicillin (82 per cent), streptomycin (3 per cent) and lincomycin (1 per cent).  相似文献   

4.
Pasteurella haemolytica biotype A serotype 1 is the principal etiologic agent of bovine pneumonic pasteurellosis. A clear understanding of the pathogenesis of this disease and the mechanisms of resistance to it has been limited by a lack of information on the important antigens of the organisms. Using recombinant DNA techniques we have cloned a segment of DNA from P. haemolytica A1 that encodes three proteins of 28, 30, and 32 kDa. Two of these proteins, 30 and 28 kDa, react strongly on a Western blot with a bovine serum raised against live cells of P. haemolytica A1. The gene for the 30 kDa protein was localized to a 3.1 kbp EcoRI fragment, and expression of the 30 kDa protein was found to be independent of an E. coli promoter. The 30 kDa protein comigrated with a 30 kDa P. haemolytica protein that was susceptible to radioiodination and presumably exposed on the bacterial cell surface. The other principal radiolabeled P. haemolytica proteins were 100, 45, and 15 kDa. Antibodies against the 30 kDa protein, isolated from E. coli carrying the recombinant plasmid, recognized 30 kDa and 15 kDa proteins in P. haemolytica serotypes 1-15 and caused agglutination of whole P. haemolytica A1 cells. Cattle vaccinated with live P. haemolytica, P. haemolytica outer membrane proteins, or the cloned 30 kDa protein developed antibodies to the cloned 30 kDa protein as detected by Western blotting and densitometry. Sera were obtained from cattle vaccinated with live or killed P. haemolytica or saline and challenged with P. haemolytica. Those sera were evaluated for antibody responses to the cloned 30 kDa protein. High antibody responses to the 30 kDa protein significantly correlated (P less than 0.01) with resistance to challenge. From these studies it is concluded that the 30 kDa protein represents a surface antigen of P. haemolytica A1 that may be important in inducing immunity to P. haemolytica.  相似文献   

5.
Mannheimia (Pasteurella) haemolytica biotype A serotype1 (A1) is the primary bacterial agent responsible for the clinical signs and pathophysiologic events in bovine pneumonic pasteurellosis. The goal of this study was to determine the prevalence of other serotypes of M. haemolytica biotype A organisms obtained from the upper Midwest diagnostic laboratories. A total of 147 M. haemolytica isolates were collected from Minnesota, South Dakota, and Michigan. Isolates were tested against M. haemolytica antisera obtained from the National Animal Disease Center, Ames, Iowa. Results indicated that M. haemolytica serotype 1 represented approximately 60%, serotype 6 represented 26%, and serotype 2 represented 7% of the total examined isolates. In addition, 7% of the isolates were serotype 9, 11, or untypable. This finding suggests that M. haemolytica serotypes other than serotype 1 can be isolated from the lung lesions of diseased cattle and seem to be capable of causing the pathologic changes observed in the lung with pneumonic pasteurellosis.  相似文献   

6.
The production and characterization of monoclonal antibodies against Pasteurella haemolytica serotype 1 is described. Ten monoclonal antibodies were produced and divided, on the basis of their properties, into six different groups. One produced bacteria agglutination only of P. haemolytica serotype 1. Three antibodies bound with P. haemolytica serotypes 1, 5-8 and 12 and the antigen was identified in immunoblots as lipopolysaccharide. Two antibodies bound P. haemolytica serotypes 1, 2, 5-8 and 12 and P. multocida serotypes 1-7, 9, 12, 15 and 16, recognizing an epitope present on a 29 kDa outer membrane protein. One antibody bound all P. haemolytica and P. multocida serotypes. The antigen was a hexosamine less than 30 kDa which contained a formalin sensitive epitope. One antibody bound only to P. haemolytica serotype 1 and the antigen was identified as a 66 kDa outer membrane protein. Two antibodies bound P. haemolytica serotypes 1, 2, 5-9 and 12 and the antigen, while not identified, was localized on the outer membrane. This study identified antigens which contribute to the cross-reactions among P. haemolytica and P. multocida serotypes and the antibodies may be useful in investigating the pathogenesis of pneumonic pasteurellosis.  相似文献   

7.
Plasmid DNA screening experiments were conducted to determine whether a relationship existed between the presence of plasmids and antibiotic resistance in Pasteurella haemolytica or the capability to produce hemolysin or leukotoxin (cytotoxin). Regardless of plasmid content, all P haemolytica isolates produced characteristic hemolysis on blood agar plates. Similarly, standardized suspensions of living bacteria and sterile concentrated (approx 200:1) culture supernatant from strains representing each of the 15 recognized P haemolytica serotypes and 7 field strains of P haemolytica (biotype A, serotype 1) produced leukotoxin, which was detected by their capability to cause inhibition of the luminol-dependent chemiluminescence response of bovine neutrophils. However, neither living bacterial suspensions nor concentrated culture supernatant from 4 untypable P haemolytica strains or a P multocida strain caused an inhibition of the luminol-dependent chemiluminescence response. The production of neither hemolysin nor leukotoxin by P haemolytica seemed to be plasmid mediated. Leukotoxin production is apparently a stable phenotypic characteristic of pathogenic P haemolytica strains, and the gene(s) coding for this activity is probably located on the bacterial host chromosome. Antibiotic susceptibility profiles were determined for the different bacterial strains. Studies of ampicillin and penicillin resistance in 8 P haemolytica (biotype A, serotype 1) strains provided evidence that the plasmid, with size of approximately 5,200 base pairs, may code for their resistance to these compounds.  相似文献   

8.
The efficacy of a multicomponent clostridial vaccine containing Pasteurella haemolytica antigens was tested in specific pathogen free or conventionally reared lambs exposed to experimental infection with P haemolytica serotypes A1, A2 or A6. In four experiments assessment was based upon the findings of clinical, pathological and bacteriological examinations. Three experiments carried out in conventionally reared lambs demonstrated protection against challenge infection with P haemolytica serotypes A1, A2 and A6 in vaccinated lambs. However, the inconsistency of the disease induced in these experiments emphasised the need to perform definitive studies in specific pathogen free conditions. The final experiment was carried out with specific pathogen free lambs and confirmed the efficacy of the multicomponent clostridial vaccine containing P haemolytica antigen in protecting against the effects of infection with P haemolytica serotype A6. In addition, this experiment indicated that the inclusion of several components in a vaccine did not affect the efficacy of an individual antigenic component.  相似文献   

9.
A protein from Pasteurella haemolytica that was highly immunogenic and toxic toward bovine alveolar macrophages was partially purified. When isolated from culture supernatants of P haemolytica serotype 1 or serotype 6, the protein reacted on Ouchterlony immunodiffusion tests with antisera from 12 serotypes of P haemolytica, but did not cross-react with antisera to serotypes of P multocida. This indicated that the protein may be specific for P haemolytica. Bacteria were grown in dialysis culture in a brain-heart infusion and calf-serum growth medium. The protein was isolated from the medium by ultrafiltration and size-exclusion chromatography and has a molecular weight of approximately 150,000 daltons. The protein, which is highly immunogenic and has the characteristics of a virulence factor, is common to all serotypes of P haemolytica, and may be an effective agent for immunization against P haemolytica in cattle.  相似文献   

10.
A luminol-dependent chemiluminescence (LDCL) assay was used to evaluate the response of bovine polymorphonuclear leukocytes; (neutrophils [PMN]) to living and heat-killed Escherichia coli, Pasteurella multocida (type A, serotype 3), and P haemolytica (biotype A, serotype 1), and to heat-killed P haemolytica and sterile culture supernatant from living P haemolytica. Control cultures containing PMN that had not been phagocytically stimulated with bacteria had a modest increase in LDCL during the initial 10 minutes of incubation, followed by a gradual decline throughout the 120-minute incubation period. Bovine PMN emitted LDCL more efficiently when the cells were exposed to living E coli or P multocida than when they were exposed to the same bacteria killed by heat. The mean LDCL values for reaction mixtures containing living E coli or P multocida peaked at 30 minutes of incubation and remained above values for mixtures containing the same heat-killed bacteria. Kinetics of the LDCL response of bovine PMN to heat-killed P haemolytica were similar (although reduced in amplitude) to that observed with killed E coli or P multocida. The LDCL response of bovine PMN to living P haemolytica was not like that for E coli or P multocida, and was characterized by the development of a peak response at 10 minutes followed by a precipitous decrease in responsiveness and a subsequent complete cessation of LDCL. Addition of sterile culture supernatant from living P haemolytica to test samples containing heat-killed P haemolytica induced a response similar to that obtained with the living microorganism.(ABSTRACT TRUNCATED AT 250 WORDS)  相似文献   

11.
OBJECTIVE: To characterize Pasteurella spp isolated from healthy pack goats and evaluate the effects of administration of a commercial Pasteurella vaccine. ANIMALS: 45 goats. PROCEDURE: Pharyngeal swab specimens and blood samples were collected on day 0 before vaccination with a Pasteurella (Mannheimia) haemolytica serotype A1 bacterin. Samples were also collected from 17 goats on days 21 and 35. Isolated Pasteurella spp were assigned to biovariant groups on the basis of results of biochemical utilization tests and serotyped. Serum antibody titers were determined. RESULTS: Multiple strains of Pasteurella spp were isolated from swab specimens and assigned to 30 nonhemolytic and 14 beta-hemolytic biovariant groups. The most common biovariant isolated was nonhemolytic P trehalosi belonging to group 2. This strain was isolated from 41 goats. Nonhemolytic P haemolytica strains were isolated from 31 goats, whereas beta-hemolytic strains of P trehalosi and P haemolytica were isolated from 8 and 35 goats, respectively. Vaccination with the A1 serotype did not affect the proportion of goats from which we isolated each biovariant group or the number of biovariant groups isolated. CONCLUSIONS AND CLINICAL RELEVANCE: Multiple strains of P haemolytica and P trehalosi belonging to nonhemolytic and beta-hemolytic biovariant groups were isolated from the pharynx of healthy domestic pack goats. Because hemolytic activity correlates with leukotoxin production, beta-hemolytic strains may have a greater potential to cause disease in naive populations of wild ruminants. However, vaccination with an A1 serotype bacterin did not decrease the proportion of culture-positive goats.  相似文献   

12.
Electrophoresis in polyacrylamide gels of the constituent proteins of the 12 serotypes and an untypable strain of Pasteurella haemolytica showed a pattern of bands that divided the group into two. This division conformed to the A and T biotype groupings of Smith (1959) although the serotype A9 showed only minor band difference from the three T serotypes 3, 4 and 10. It was not possible by this method to separate all the type strains from each other by the specific recognition of the patterns of protein mobilities produced.  相似文献   

13.
A significant obstacle to molecular studies of Mannheimia (Pasteurella) haemolytica, has been its resistance to genetic transformation. The lack of competence of many M. haemolytica strains has been attributed to the presence of restriction modification systems. In this study, representative strains of 12 M. haemolytica serotypes and four Pasteurella trehalosi serotypes were successfully transformed by electroporation using a recombinant vector derived from the native M. haemolytica A1 serotype plasmid pNSF2176. Transformation was achieved despite PCR-based evidence for the presence of genes encoding a type I restriction enzyme, phaI, and a type II restriction enzyme hsdM, in each of the M. haemolytica strains.  相似文献   

14.
Live Pasteurella haemolytica biotype A, serotype 1 isolates (n = 3) and Escherichia coli K-12, strain W3110, were reacted with bovine pulmonary lavage cell (PLC) suspensions. The comparative effects of the different bacteria on the functional and metabolic activity of alveolar macrophages (AMO) in the PLC suspensions were assessed simultaneously by use of 51Cr release, luminol-dependent chemiluminescence (LDCL), and AMO bactericidal assays. The bovine PLC responded differently to E coli, than to the 3 P haemolytica isolates in each of the 3 experimental test systems; however, responses to each of the P haemolytica isolates were not found to be significantly different. Unopsonized live P haemolytica cells adversely affected the functional and metabolic response of PLC, whereas there was no evidence of a cytotoxic (cytocidal) influence of E coli. A difference in 51Cr release for reaction mixtures containing E coli and P haemolytica was not detected at zero time; however, at each subsequent time, reaction mixtures phagocytically stimulated with P haemolytica had significantly increased amount of 51Cr release (P less than 0.05), compared with those mixtures containing E coli. Bovine AMO in the PLC suspensions were able to effectively kill E coli in vitro, but were unable to prevent survival and subsequent growth of P haemolytica. The luminol-dependent chemiluminescence profiles for reaction mixtures phagocytically stimulated with E coli provided evidence of sustained production of oxygen radicals with antimicrobial capabilities by bovine AMO in the PLC. Production of these highly reactive antimicrobial oxidants appeared initially in cultures containing P haemolytica but, subsequently, their production declined precipitously and ceased altogether.  相似文献   

15.
Experimental immunisation of lambs against pneumonic pasteurellosis   总被引：1，自引：0，他引：1  
Methods of immunising lambs against pneumonic pasteurellosis, caused by several serotypes of Pasteurella haemolytica, were assessed in specific pathogen free lambs. Lambs were vaccinated intramuscularly with sodium salicylate extract (SSE) of P haemolytica, either alone or in combination with heat-killed organisms (HKO). SSE of P haemolytica type A1 protected vaccinated lambs against pneumonia resulting from challenge with the homologous serotype. SSE of type A2 also provided some protection but this was improved by vaccination with a combination of SSE and HKO.  相似文献   

16.
In three experiments subcutaneous vaccination of calves with adjuvanted bacteria-free leukotoxic culture supernatant from log phase cultures of Pasteurella haemolytica A1 (toxin 1) was shown to induce some protection against intrabronchial challenge with live P. haemolytica A1. This toxin 1 vaccine was as effective as a whole cell bacterin in stimulating agglutinating antibody to P. haemolytica. Induction of leukotoxin neutralizing activity was variable; in some cases vaccination only primed the animal to produce an anamnestic response after challenge, whereas in other instances antitoxic activity increased in response to immunization. Two doses of vaccine were shown to be more effective than a single immunization. Vaccination with leukotoxic culture supernatant from the nonpathogenic P. haemolytica serotype 11 was as effective as vaccination with toxin 1 in stimulating antitoxic activity but was not protective. This implies that both serospecific agglutinating activity and an antitoxic response are needed for immunity.  相似文献   

17.
Vaccination of calves with a Pasteurella haemolytica serotype 1 antigen preparation elicited a serotype-specific inhibition of nasal colonization by P haemolytica under field conditions. Inhibition was evidenced by a low frequency of nasal colonizations and by relatively few P haemolytica serotype 1 organisms isolated from vaccinated calves. The study comprised 3 field trials, each on a separate year, and included 480 calves.  相似文献   

18.
Several modern electron microscopy techniques were used to examine Pasteurella haemolytica (biotype A, serotype 1) (strain B122) recovered from experimentally infected cattle and in situ within the lung tissue of experimentally infected cattle. Glycocalyx four to five times thicker than that seen on P. haemolytica grown in vitro was evident on bacterial cells recovered from live infected calves by pulmonary lavage. Fimbriae were also present on cells recovered by lavage. A thick glycocalyx was also seen on P. haemolytica-A1 within the lungs of experimentally infected cattle at necropsy. In summary, cells of P. haemolytica-A1 in experimentally infected cattle have fimbriae and glycocalyx on their cell surfaces and these structures appear to be important in bacterial colonization of the bovine respiratory tract and pathogenesis of shipping fever (Pasteurella) pneumonia.  相似文献   

19.
Serological types of Pasteurella haemolytica in Kenya   总被引：1，自引：0，他引：1  
The 12 known serotypes of P. haemolytica and two biotypes A and T were found to occur in Kenya. Biotype T was more common in disease conditions than biotype A, which was more common in the nasal passages of healthy animals. Only biotypes A strains were recovered from cattle and the majority were serotypes 1 and 2, but serotypes 4 and 11 were also isolated. All serotypes were found to occur in sheep and goats, but serotypes 3, 4, 6, 8 and 10 were more commonly associated with pneumonia. It was observed that chickens could harbour both biotypes A and T in pathological conditions. Biotype A serotype 2 was isolated from an adult wildebeest, but the prevalence of P. haemolytica in wild animals needs furter investigation.  相似文献   

20.
Hybridoma-derived monoclonal antibodies (MAB) against the cell surface antigens of Pasteurella haemolytica serotype 1 were obtained by the fusion of murine myeloma cells (P3 X 63 - Ag 8.653) with splenocytes of BALB/c mice immunized with crude logarithmic growth-phase culture supernatant. Initial screening was performed, using an ELISA, with the same bacterial growth culture supernatant as coating antigens. Further selection was done, using a panel of purified antigens--either capsular polysaccharide or lipopolysaccharide--as the coating antigen in an ELISA, and then performing a leukotoxin-neutralization assay. Two MAB, designated IIB-6 and H-2, reacted specifically with the capsular polysaccharide and the other 3, designated IVG-3, IH-3, and IIC-2, reacted with the lipopolysaccharide. One MAB, designated IH-6, did not react with leukotoxin, capsular polysaccharide, or lipopolysaccharide. The MAB to the capsular polysaccharide (IIB-6 and H-2) were characterized further; both antibodies belonged to the IgM class and were agglutinating. In addition, they promoted neutrophil-mediated opsonophagocytosis and complement-mediated immune bacteriolysis of P haemolytica serotype 1. Results from 3 studies indicated that the MAB IIB-6 and H-2 were specific only to the capsular polysaccharide of serotype 1 of P haemolytica. The MAB to the lipopolysaccharide (IVG-3, IH-3, and IIC-2) were of the IgG1, IgG3, and IgM classes, respectively and were not characterized further. The availability of a MAB identifying a serotype-specific, surface-exposed determinant on the capsule of P haemolytica serotype 1 should facilitate and expand studies concerning the role of the capsular material and lipopolysaccharide in the pathogenicity of P haemolytica infection in cattle.  相似文献   

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