Antimicrobial susceptibility of Pasteurella multocida isolated from cattle and pigs

Minimum inhibitory concentrations (MICs) of 10 antimicrobial agents were determined for Pasteurella multocida from cattle and pigs (72 and 68 isolates, respectively). Higher MICs were observed with oxytetracycline, doxycycline, tilmicosin and thiamphenicol for porcine isolates than for bovine isolates. Enrofloxacin was the most active, with an MIC for 90% of the isolates (MIC90) of 0.05 microg/ml for both bovine and porcine isolates. Aspoxicillin exhibited the same excellent activity against penicillin-susceptible isolates as ceftiofur, with MICs ranging from < or = 0.025 to 0.1 microg/ml. Aminoglycosides were less active, with an MIC90 of > 100 microg/ml for both bovine and porcine isolates.     

Association of complement sensitivity with virulence of Pasteurella multocida isolated from turkeys

Two strains of Pasteurella multocida, both derivatives of strain P1059, were compared for virulence for 14-week-old turkeys and sensitivity to turkey plasma. Strain P1059-1, a nalidixic-acid-resistant mutant of P1059 with an LD50 of approximately 10(3) colony-forming units (CFU), was more resistant to the bactericidal effects of fresh turkey plasma at 37 C than avirulent strain P1059-1A. P1059-1A, with an LD50 of approximately 10(8) CFU, is an acapsular variant of P1059-1 that spontaneously arose after prolonged passage on artificial medium. The bactericidal effect on P1059-1A was removed when turkey plasma was treated with heat or with zymosan, maneuvers that removed hemolytic complement activity from turkey plasma.     

Genetic diversity and associated pathology of Pasteurella multocida isolated from porcine pneumonia

Pasteurella multocida is a widespread respiratory pathogen in pigs associated with atrophic rhinitis and contributing to aggravation of the pulmonary lesions. The aims of the present study were to characterize isolates of P. multocida from porcine bronchopneumonia by pulsed-field gel electrophoresis (PFGE), PCR based capsular typing and multilocus sequence typing (MLST) and to compare clonal complexes outlined with the type of histological lung lesions to investigate if a correlation between clonal lineages and lesions might exist. Isolates of P. multocida were obtained from cases of cranioventrally located porcine bronchopneumonia. All lung lesions were described and classified according to histological lesions. A total of 139 isolates, from lung (n=111), pericardial sac (n=21) and kidney (n=7) of 111 pigs were described using PFGE with ApaI as the restriction enzyme. Furthermore, 20 and 29 isolates were characterized by capsular serotyping and multilocus sequence typing, respectively. PFGE demonstrated 15 different clusters showing 50% or more similarity. All selected isolates were of capsular serotype A and only three main sequence types (ST) were detected among the isolates. Associations were not found between histopathology and clonal complexes of P. multocida. In conclusion, PFGE demonstrated a high diversity of genotypes of P. multocida associated with porcine bronchopneumonia. However, isolates obtained mainly belonged to few STs, indicating that isolates of P. multocida associated with porcine bronchopneumonia originates from a limited number of clonal lineages and therefore might have adapted to porcine hosts. No correlation was demonstrated between genotypes and types of lesions, and extra-pulmonary spreading was only rarely demonstrated.     

Biochemical and serological studies on Pasteurella multocida isolated from cattle in the Sudan.

Of 42 strains of Pasteurella meltocida isolated from different outbreaks of hemorrhagic septicaemia and from healthy cattle in various parts of the Suden, 38 belonged to Carters' type B and four to type E. The strain used for vaccine production was type E. With respect to somatic antigen determination, there was some correlation between the results of the gel diffusion and the agglutination tests as most strains reacted with 0 groups 6 antisera in both tests. However, some antigens cross-reacted with antisera of different 0 groups in the gel diffusion test. No correlation was observed between the pattern of biochemical reactions and the serological types.     

Resistance plasmids of Pasteurella multocida isolated from turkeys

From 1940 through 1978, fifty-eight strains of Pasteurella multocida (serotype 3) were isolated from turkeys throughout the United States and were examined for R-plasmids. Forty-one of the isolates contained plasmid DNA, of which 7 isolates were found to encode resistance to tetracycline, streptomycin, and sulfonamides, or to streptomycin and sulfonamides. The R-plasmids were 2 to 10 megadaltons, nonconjugal, and contained a moles percent guanine plus cytosine ratio in the range of 57 to 61. The R-plasmids did not belong to any of the 19 incompatibility groups evaluated, including Inc Q. Digestion with restriction endonuclease indicated that 2 of the plasmids from P multocida isolated in 1960 and 1962 were identical, whereas 4 of the 5 plasmids obtained from P multocida isolated after 1966 were identical, with the 5th plasmid closely related to the other 4. The results indicated that R-plasmids were not widely dispersed among P multocida (serotype 3) isolated from turkeys in the United States. The nontransmissible nature of these plasmids was probably the major reason for their lack of dissemination.     

Associations between biovar and virulence factor genes in Pasteurella multocida isolates from pigs in Spain

Two hundred and five isolates of Pasteurella multocida from pigs were phenotypically and genetically characterised by determining their biovar, capsular type, virulence-associated genes and pulsed-field gel electrophoresis (PFGE) profiles. All isolates were identified as P multocida subspecies multocida and most were assigned to biovar 3 (58 per cent) and biovar 2 (39.5 per cent). Biovar 1 represented 2.4 per cent of the isolates. According to the capsular type, the great majority of the isolates (79.0 per cent) belonged to capsular type A, 18.5 per cent belonged to capsular type D and 2.4 per cent were of capsular type F. All isolates harboured ompH, psl, oma87, ptfA, nanB, nanH, tonB, hgbA, sodA and sodC genes, while none of them possessed the transferrin-binding protein gene tbpA. The prevalence of toxA, pfhaA and hgbB genes was variable (7.8, 40.5 and 60.5 per cent of the isolates, respectively). After PFGE typing, isolates of biovar 2 and 3 were grouped in two different clusters (A and B) at a level of 45 per cent similarity. In addition, isolates of biovar 2 and 3 exhibited statistically significant differences (P<0.05) in the virulence-associated hgbB and pfhA genes (biovar 3 was hgbB(+) pfhA(-), while biovar 2 was hgbB- pfhA(+)).     

Characterization of Pasteurella multocida mutants of low virulence

Ten temperature-sensitive mutants of the Clemson University (CU) vaccine strain of Pasteurella multocida have been developed and were characterized by phenotypic attributes such as carbohydrate fermentation, antibiotic resistance, and membrane protein profiles. Some mutants were found to have lost the ability to utilize some substrates, notably xylose and gluconate, whereas others were able to ferment additional carbohydrates such as arabinose and rhamnose. CU was found to be resistant to sulfisoxazole, of intermediate resistance to bacitracin, and sensitive to rifampin; the sensitivity to these three antibiotics varied among the mutant strains, but 60% were resistant to rifampin. Membrane protein profiles demonstrated some changes in major bands, and there was variation in 50% of the mutants in proteins in the 31 kilodalton range. All strains were assayed for the presence of several virulence factors, and many were found to produce siderophore and to exhibit some degree of complement resistance.     

Characterization of Pasteurella multocida strains isolated from human infections

Isolates of Pasteurella multocida recovered from infected humans (n = 15) were characterized by traditional and molecular microbiological methods and were compared with cat-derived strains (n = 5). The most prevalent subspecies among strains from human infections was P. multocida subsp. septica (80%), and nearly all isolates showed a similar combination of virulence-associated genes. MLST analysis classified the 20 P. multocida strains into 16 different sequence types, and we assigned 11 new sequence types (ST), however, only one of those (ST 334) was shared by two human and one cat isolates. P. multocida subsp. septica strains formed a distinct phylogenetic group within the species. The strains showed resistance to erythromycin, clindamycin and sulfamethoxazole, and with two exceptions, resistance to tilmicosin was also detected. Each strain was susceptible to ampicillin, streptomycin, gentamycin, tetracycline, doxycycline, cefazolin, cefpodoxime, chloramphenicol, florfenicol and enrofloxacin. Common characteristics (virulence profile and antibiotic sensitivity pattern) shared by strains isolated from humans and cats support the view that domestic cats may serve as a potential reservoir for P. multocida.     

A serotypic survey of Pasteurella multocida isolated from poultry

One hundred forty-eight Pasteurella multocida isolates from four southeastern states and California were serotyped by a gel diffusion precipitin test. The isolates were predominantly from turkeys and chickens. Sixty-eight percent of the isolates had antigenic characteristics of serotypes 3 and 4 (3 X 4). In turkeys, 76% of the isolates were 3 X 4, and serotype 3 was second (17%) in frequency. In chickens, 54% of the isolates were 3 X 4 and 19% were serotype 1.     

A survey of potential virulence markers from avian strains of Pasteurella multocida

Twenty-four isolates of Pasteurella multocida from clinical cases of fowl cholera and the Clemson University vaccine strain were surveyed for the presence of potential virulence markers. Membrane proteins, enzymatic activity of the membrane proteins, and carbohydrate fermentation patterns were also determined to demonstrate phenotypic relationships within the groups. Few differences were found in these phenotypic characteristics among the isolates. Almost all the organisms produced siderophore and were hemolytic on turkey red blood cells. No extracellular enzyme or bacteriocin activity was detected and little antibiotic resistance was found. However, many organisms contained plasmids and demonstrated some degree of resistance to complement. Both characteristics were correlative markers in Pasteurella multocida isolated from birds with fowl cholera.     

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

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

Characterization of Pasteurella multocida isolated from rabbits in Canada.

In a survey for the somatic and capsular serotypes of Pasteurella multocida present in domestic rabbits in Canada, but mainly in Ontario, samples were obtained from research facilities, commercial rabbitries and from abattoir and necropsy specimens. Sources of isolates were upper respiratory tract infections, localized bronchopneumonias , acute fibrinous pneumonias, abscesses and otitis media. Of 59 isolates obtained, 47.0% were type 12:A, 30.5% 3:D and 12.0% were 3:A. Less common types were 12(4):A, 12:D, 4(12):A and 3:untypable. Somatic group 3 was most commonly isolated from acute pneumonic disease, while serogroup 12:A was most commonly found in upper respiratory tract infections and in localized chronic bronchopneumonia. Two serotypes of P. multocida were isolated from four pneumonic lungs collected from abattoir specimens. Most isolates were susceptible to the commonly used antibiotics.     

Rapid virulence typing of Pasteurella multocida by multiplex PCR

The gram-negative bacterium Pasteurella multocida constitutes a heterogeneous species associated with wide range of disease in many animals. Isolates are classified into five groups based on capsular antigen (capA, B, D, E and F). Recently, a new valuable PCR-based method was introduced to determine the epidemiological correlation between P. multocida infection and existence of virulence genes including tbpA, pfhA, toxA and hgbB. However, this method is tedious and laborious. Thus, in the current study, we designed a reliable multiplex PCR method for rapid detection of virulence genes in P. multocida. Eighty seven strains of P. multocida isolated from various clinically healthy and infected hosts were examined by uniplex PCR method for each virulence associated genes. Based on our improved and simplified multiplex PCR method, rapid detection of four virulence genes was accomplished. It is proposed that its implementation may benefit the epidemiological investigations.     

Capsular serogroups of Pasteurella multocida isolated from animals in Zimbabwe

Pasteurella multocida is isolated from a variety of disease conditions from different animal species in our diagnostic laboratory. In order to determine serogroup distribution among the isolates, an indirect haemagglutination test using glutaraldehyde-fixed sheep red blood cells was employed. A serological examination of 79 isolates revealed that 47/79 were of capsular serogroup A, 11/79 capsular serogroup D, 4/79 capsular serogroup B and 17/79 were untypable strains. None of the isolates belonged to either serogroup E or F. All those from cases of classical pasteurellosis could be grouped, but a significantly high proportion of those which originated from companion animals were untypable. The significance of these results is discussed. This report appears to be the first detailed information on the prevalence of various serogroups of P. multocida in animals in southern Africa.     

Pasteurella multocida recovered from live turkeys: prevalence and virulence in turkeys

Swabs of the oropharynges of 801 live turkeys (621 meat birds and 180 breeders), collected from 15 flocks that had experienced an outbreak of fowl cholera and from 12 non-outbreak flocks, were screened for the presence of Pasteurella multocida. Turkeys from outbreak flocks were sampled within 2 to 9 weeks of the outbreak. Forty-nine isolates of P. multocida were recovered from turkeys in 11 of the outbreak flocks, and none were recovered from turkeys in non-outbreak flocks. Isolation rates varied from 0 to 72% of turkeys sampled in a flock. Nineteen isolates were tested for virulence by injecting them intravenously into turkeys, and 14 were lethal. Results demonstrated that for purposes of disease control, meat birds in fowl-cholera-outbreak flocks should be considered carriers of potentially virulent P. multocida for the life of the flock.     

Characterization of outer membrane protein-enriched extracts from Pasteurella multocida isolated from turkeys

Outer membrane protein (OMP)-enriched extracts of avian strains of Pasteurella multocida were examined by use of sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Culture medium did not have a significant effect on the OMP profiles of strains of P multocida examined; however, in vivo propagation had an appreciable effect on the OMP profile composition of the reference strain P-1059. Such bacteria, expressed several additional OMP in the 27-kD, 48-kD, 56-kD, 60-kD, 80-kD, and 94-kD molecular mass regions. These OMP were not detected in the electrophorogram of strain P-1059 grown in vitro. The OMP profiles of reference strains of the 16 serotypes of P multocida did not identify any serotype-specific protein markers. Field strains of serotype A:3 had variation in OMP profiles and did not express OMP that all were identical to that expressed by the reference strain P-1059. The live attenuated CU and M9 bacterial vaccine strains expressed strain-specific OMP markers of 48-kD and 45-kD molecular masses, respectively. These strain-specific OMP markers may be used to differentiate these strains from virulent field strains that are of the same serotype and isolated from turkeys that have succumbed to pasteurellosis as a result of vaccine-related reactions or breakdown in immunity.