Experimental induction of pneumonic pasteurellosis in calves by intratracheal infection with Pasteurella multocida biotype A:3

The study aimed to establish an experimental model to investigate the pathogenesis of lung infection by Pasteurella multocida, an important cause of bovine respiratory disease. An experimental model is required to assist the development of an effective vaccine. Sixteen 8-week-old calves were challenged intratracheally with 10(9) or 10(10) colony forming units of P. multocida in either 60 or 300 ml saline in a 2 x 2 factorial experiment. All animals became dull within 2-6h post-infection (p.i.) and two calves were killed humanely because of suspected endotoxic shock. Remaining animals showed increased respiratory rates by 15-20 h p.i. and, at 23 h p.i., calves given the high dose, high volume challenge showed higher (P < 0.05) rectal temperatures. From 24 to 36 h p.i., clinical signs decreased in a majority of animals. Plasma haptoglobin concentrations increased (P < 0.05) in calves given the high volume challenge irrespective of the number of bacteria. At post-mortem examination (4d p.i.), lung lesions, mainly in the apical lobes, were found in all calves. Histopathological examination showed areas of purulent pneumonia with a tendency to abscessation and inflamed interlobular septa characterised by accumulation of neutrophils and oedema. The clinical and pathological responses described were typical of bovine pneumonic pasteurellosis.     

Experimental Pasteurella multocida pneumonia in calves

Pasteurella multocida was isolated from the lungs of calves that died on a farm in the south of England. This organism was inoculated experimentally into 13 calves by the intratracheal route: in all but two of the calves mild clinical disease resulted and at necropsy, three or four days later, pneumonic consolidation involving up to 22 per cent of the lung was observed. P multocida was isolated from all but two of the lungs. Of two calves inoculated intravenously with P multocida, one showed mild clinical disease and slight pneumonic consolidation at necropsy and the other remained normal. Control calves inoculated intratracheally and intravenously with sterile broth showed no signs of illness and no pneumonic consolidation. Histologically the lung lesions comprised a fibrinous bronchopneumonia with variable sized areas of coagulative necrosis, extensive deposition of fibrin and massive dilatation and oedema of the interlobular and pleural lymphatics. It is concluded that P multocida should receive more recognition as a primary pathogen.     

Pathological and immunological changes after challenge infection with Pasteurella multocida in naive and immunized calves

Challenge infections of calves with Pasteurella multocida were established to characterize the local inflammatory response and determine the effect of previous exposure to live bacteria on the post-challenge immune response. Experimental infections were established by intratracheal inoculation of P. multocida in both naive calves and calves that had been previously vaccinated with two subcutaneous (s.c.) injections of live bacteria. Histological, immunohistological and cytokine expression studies were performed on bronchoalveolar lavage (BAL) samples, lung parenchymal tissues and lung lymph nodes (LN). In comparison to uninfected control animals in which no lung lesions were observed, a patchy to confluent bronchopneumonia was observed following infection of naive calves, characterized by abscess formation, haemorrhage, oedema and suppurative consolidation. Cellular analysis following infection of naive animals was characterized by an influx of neutrophils in the BAL, with macrophages and dendritic cells observed in the lesion perimeter. A significant increase in the number of CD8(+) blasts expressing MHC (major histocompatibility) II was also observed in the BAL of infected calves. Decreased expression of interleukin (IL)-1 beta and increased expression of IL-8 compared to naive unchallenged controls was apparent in lung LN. In comparison, a more limited pathology was observed in vaccinated animals post-challenge, indicating partial protection conferred by the s.c. immunization with live bacteria. Studies of vaccinated animals showed the presence of bronchial-associated lymphoid tissue (BALT) in the lung tissue and an increase in the number of B-cells and CD4(+) T-cells expressing MHCII in the lung LN after challenge. In contrast to primary infection, there was no significant influx of neutrophils in the BAL. Instead, a population of newly recruited monocytes/macrophages was observed. Increased IL-2 expression and decreased IL-8 expression was observed in the LN, while IL-1 beta expression was not detected. The reduced neutrophil and increase monocyte response in the vaccinated calves may be associated with significant changes in the gamma delta T lymphocyte population in the BAL.     

Pasteurella multocida infection in heterophil-depleted chickens

The present study was aimed at elucidating the role of heterophil granulocytes during the initial infection with Pasteurella multocida subsp. multocida in chickens. Chickens (17 and 19 wk old) were depleted of their heterophil granulocytes by 5-fluorouracil treatment. When the heterophil blood counts were significantly reduced, the birds were inoculated intratracheally with 1.8-4.3 x 10(4) colony-forming units of P. multocida. Twelve, 24, or 48 hr postinoculation, the birds were euthanatized and examined for macroscopic and histologic lesions in the lungs. Bacterial invasion was determined by culture of P. multocida from the spleen. Recruitment of heterophils into the respiratory tract during infection was found to contribute considerably to the lung lesions in chickens and was found to mediate tissue damage, possibly allowing a more rapid systemic spread of P. multocida. However, during progression of the infection, the heterophil-mediated necrosis in chickens seemed to stimulate giant cell demarcation of infected lung tissue, which coincided with the clearance of P. multocida from the spleen, thus hampering further invasion. Consequently, heterophil activation plays a dual role for the outcome of a P. multocida infection in chickens, where it initially seems to promote invasion and systemic spread but subsequently helps limit the infection by giant cell formation and bacterial clearance.     

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.     

Molecular epidemiology of Pasteurella multocida in dairy and beef calves

The molecular epidemiology of Pasteurella multocida has rarely been studied at the farm level in cattle. The aim of this study was to determine whether single or multiple strains of P. multocida tend to exist within farms. Molecular characterisation was carried out on isolates obtained from nasal swabs from 105 calves from 32 randomly selected beef and dairy farms located throughout Scotland, and from 131 calves from 20 farms in the Mayenne region of France, where sampling occurred in response to respiratory disease outbreaks. P. multocida isolates were characterised by random-amplified polymorphic DNA (RAPD) typing and pulsed-field gel electrophoresis (PFGE) using restriction enzyme ApaI. In addition, isolates representative of each farm/RAPD profile combination were typed by multilocus sequence typing (MLST). Among 105 Scottish isolates, 15 RAPD profiles were distinguished. The majority of farms (27/32) had indistinguishable profiles in all positive animals. Five farms had two profiles. Among 140 French isolates, 23 RAPD profiles were distinguished. More within-farm heterogeneity was observed although 10/20 farms had just one profile (E4) in sampled calves. Profile E4 accounted for 60% (84/140) of French isolates. PFGE was more discriminatory than RAPD but confirmed results with respect to within farm homogeneity or heterogeneity of strains, whereas MLST was not discriminatory enough for farm level epidemiology. As in other host species, either several strains or one dominant strain of P. multocida may exist within farms, with evidence for a role of management factors such as movements onto the farm in the number of strains detected.     

Polypeptides associated with Pasteurella multocida infection in rabbits.

Polypeptides from whole cell preparations of Pasteurella multocida serotypes A:12 and A:3 were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and transferred to nitrocellulose paper. Antigens were detected by immunoblot analysis, using sera from 3 groups of rabbits. Sera were obtained from rabbits inoculated intranasally with P multocida serotype A:12 or A:3, from rabbits maintained in a rabbitry with enzootic P multocida A:12 infection, and from rabbits maintained in a rabbitry with enzootic P multocida A:3 infection. Immunoblot analyses of pre- and postinoculation sera from experimentally infected rabbits, using serotype A:12 antigen, revealed 3 polypeptides with approximate molecular mass of 28, 30, and 37 kDa that consistently detected antibodies after P multocida-induced infection. Sera from rabbits naturally infected with either serotype, tested against serotype A:12 and A:3 antigens, detected the same polypeptides in both serotypes. Thus, immunologic reactivity to these polypeptides may be useful for serologic detection of P multocida infection.     

Immunoperoxidase evaluation of pneumonic lesions induced by Pasteurella multocida in calves

To evaluate the relationship between pneumonic lesions and distribution of bacteria, lungs from calves inoculated with Pasteurella multocida were examined histologically by use of immunoperoxidase technique. Pneumonic lesions fundamentally consisted of broncho-pneumonia with fibrinopurulent pleuritis. The lesions were confirmed to be associated with inoculated P multocida, using the immunoperoxidase technique. The P multocida antigen was detected not only in the bacterial clusters in the lesions, but also in the cytoplasm of infiltrating neutrophils and macrophages. Further, immunoelectron microscopy indicated that the inoculated bacteria generally were phagocytosed and digested by neutrophils.     

Experimental transmission of Pasteurella multocida 6:B in goats

Haemorrhagic septicaemia (HS) is an acute disease of cattle and buffaloes caused by Pasteurella multocida 6:B. Outbreaks of the disease have been closely associated with carrier animals that transmit the organism to susceptible animals during stressful condition. This study was conducted to determine whether goats exposed intranasally to P. multocida 6:B can transmit the organism to contact goats. Thirty-six healthy local Katjang goats were divided into four groups and goats of groups 1 and 3 were each inoculated intranasally with a 1-ml inoculum that contained 1 x 10(9) CFU/ml of live P. multocida 6:B. Following the exposure, all goats of groups 3 and 4 were injected with dexamethasone at the rate of 1 mg/kg for three consecutive days. At the end of the dexamethasone treatment, goats of groups 1 and 2 were commingled but kept separate from goats of groups 3 and 4, which were commingled in another pen. Three surviving goats from each group were killed on days 7, 14 and 21 post-exposure for postmortem examination. Naso-pharyngeal mucus and heart blood were collected on swabs. Tissues from lungs, lymph nodes and tonsils were collected for bacteriological isolation and identification. Only one goat of group 3 died 6 days post-exposure showing clinical signs and lesions typical of HS. Other goats showed mild signs of upper respiratory tract infection. Goats of all groups developed acute mild pneumonic lesions, however, those treated with dexamethasone had significantly (P < 0.05) more extensive lesion scoring based on the lesion scoring system. P. multocida 6:B was isolated from the nasal mucosa and lung lesions of exposed and contact goats not treated with dexamethasone. Exposed and contact goats treated with dexamethasone carried the organism for 21 days. P. multocida isolation from heart blood was made only from exposed and contact goats treated with dexamethasone. P. multocida was isolated from the lymph node of the goat that died during the experiment.     

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.     

麝鼠巴氏杆菌分离鉴定及其对组织病理性损伤

巴氏杆菌病由多杀性巴氏杆菌所引起的家畜、野禽和野生动物共患的一种以败血症和出血性炎症为特征的急性传染病,其发病率和死亡率很高,严重危害动物养殖业.     

Immunologic and physiologic responses of calves inoculated with potassium thiocyanate extract of Pasteurella multocida type A

Inoculation of calves with potassium thiocyanate (KSCN) extract of Pasteurella multocida type A in saline-tris buffer or in Freund's incomplete adjuvant or modified Freund's incomplete adjuvant resulted in the elicitation of agglutinating, hemagglutinating, bactericidal and homocytotropic antibodies. The antibody response was significantly (P = 0.05) higher in calves inoculated with the KSCN extract in either adjuvant than in those inoculated with the extract in saline-tris buffer. Hemagglutination also was observed if the KSCN extract of P hemolytica was used for sensitizing the tanned sheep red blood cells. Further, calf anti-P multocida extract antisera also was bactericidal to P hemolytica. The KSCN extract of P multocida was found to be nontoxic to calves at 2 doses tested, as judged by an evaluation of total and differential leukocyte counts, body temperature, and pulse rates at various intervals after inoculation.     

Experimental Pasteurella multocida infection of swine. A contribution to the etiology of enzootic porcine pneumonia

Pasteurella can be isolated from about 50 per cent of pneumonically changed pig lungs. Isolation, however, is possible also from intact lungs of conventional or secondary SPF animals. The authors of this paper used the positive hyaluronidase test and identified exclusively Pasteurella multocida, with predominance of capsular type A. Most of the Pasteurella strains tested in this context caused pneumonia in SPF animals, particularly, following intratracheal application. No correlations were found to exist with murine virulence, the serological type or the origin of the strains from affected or intact lungs. Apart from virulence differences between the strains, host reactivity was found to depend primarily on high germ counts and, under natural conditions, on environmental stresses. Pasteurella multocida, hence, can be considered as one of the potential pathogens of porcine pneumonia. Pneumonia foci with transduced localised fibrinous pleuritis are characteristic and constitute a process of purulent inflammation with early involvement of fibrin.