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Biomedical subjects

H A Gibbs

Publications and source records attributed to H A Gibbs.

36 records · Page 2Linked to original sources

Outer-membrane protein and lipopolysaccharide variation in Pasteurella haemolytica serotype A1 under different growth conditions.

Growth characteristics, as well as outer-membrane protein (OMP) and lipopolysaccharide (LPS) profiles in SDS-polyacrylamide gels, of two serotype A1 isolates of Pasteurella haemolytica were examined under different in vitro growth conditions. The two isolates were chosen as representatives of disease (S/C 82/1) and non-disease (W/D 83/4) isolates, respectively. The growth rates and final cell densities of both isolates increased as the degree of aeration increased. In particular, the final cell densities varied significantly according to the degree of aeration. Under anaerobic conditions, however, both the growth rate and final cell density were significantly reduced. There was reduced expression of a 40.5 kDa protein under anaerobic conditions in both isolates, whereas in S/C 82/1 expression of the 71, 77 and 100 kDa iron-regulated proteins increased as aeration decreased. There were also differences in low-molecular-mass components of LPS between cells grown anaerobically and those grown aerobically. Growth in the presence of 5% CO2 did not significantly alter the growth rate and had little, if any, affect on OMPs or LPS. Differences in the expression of certain proteins occurred as growth progressed from the exponential to the stationary phase. Growth in the presence of the iron chelators 2,2'-dipyridyl, ethylenediamine-dihydroxyphenylacetic acid (EDDA), desferrioxamine mesylate (desferal), ovotransferrin (conalbumin) and bovine transferrin was inhibited within a very narrow concentration range. In the presence of 2,2'-dipyridyl, EDDA or desferal, 71 and 100 kDa iron-regulated OMPs increased in both isolates whereas a 77 kDa protein increased in isolate S/C 82/1 only. In the presence of ovotransferrin or bovine transferrin there was, in both isolates, increased expression of the 71 kDa protein, a slight increase in expression of the 100 kDa protein but no expression of the 77 kDa protein; there was also increased production of the 40.5 kDa protein, and synthesis of two additional proteins of 23 and 26 kDa. Other differences occurred after growth in foetal and newborn calf sera. In foetal calf serum there was enhanced expression of the 71 but not of the 100 kDa protein. In newborn calf serum there was no enhanced expression of the 71, 77 or 100 kDa proteins, but expression of novel proteins of 97 and 98 kDa as well as a high-molecular-mass protein occurred. There was also slight quantitative differences in the LPS profiles of cells grown in foetal or newborn calf sera compared to those of cells grown in other media.(ABSTRACT TRUNCATED AT 400 WORDS)

Aerobiosis↗

A colourimetric, microplate assay for the leucotoxin of Pasteurella haemolytica.

Culture supernates of Pasteurella haemolytica, which contain leucotoxin, inhibited the reduction of nitroblue tetrazolium (NBT) by bovine and ovine but not rabbit leucocytes in response to phorbol 12-myristate 13-acetate (PMA). Culture supernates of P. multocida, which contain no leucotoxin, had no inhibitory effect on the response of leucocytes from any species. The inhibition of NBT reduction was assessed visually or spectrophotometrically in the wells of microplates and used as a simple assay for leucotoxin. It was as sensitive as the trypan blue dye-exclusion method and did not require the use of radioisotopes. In addition, sera from P. haemolytica-infected calves inhibited leucotoxin activity in the microplate assay. Thus, inhibition of NBT reduction after stimulation of ruminant leucocytes with PMA can be used as a simple, specific assay for leucotoxin and for anti-leucotoxin antibodies.

Animals↗

Distribution of Pasteurella haemolytica in the respiratory tracts of carrier calves and those subsequently infected experimentally with Dictyocaulus viviparus.

Distribution of Pasteurella haemolytica in the respiratory tracts of calves with no apparent clinical signs of illness and those infected experimentally with Dictyocaulus viviparus was determined so as to define carrier sites for this organism. The calves had been positive by nasopharyngeal swab for either P haemolytica A2 or A1 for at least two months or for over a month, respectively, before slaughter. P haemolytica A1 was acquired following horizontal spread from other infected calves. It was observed post mortem that P haemolytica A1 or A2 resided in the tonsils and retropharyngeal lymph nodes of calves of both groups. In addition to these sites, P haemolytica A1 was also isolated from the right cranial lung lobe of one of the calves from the D viviparus infected group although there was no evidence of pasteurella associated pneumonia. It was concluded that tonsil and retropharyngeal lymph nodes appear to be the most important carrier sites for P haemolytica when compared to other tissues of the bovine respiratory tract.

Animals↗

Pasteurella species isolated from the bovine respiratory tract and their antimicrobial sensitivity patterns.

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).

Animals↗

Sequential lesions of experimental bovine pneumonic pasteurellosis.

A strain of Pasteurella haemolytica biotype A serotype 1, which had been isolated from a pathologically-confirmed outbreak of bovine pneumonic pasteurellosis, was used successfully to reproduce the disease in conventional calves. The development of the various pathological features was studied at regular intervals following infection. The acute inflammatory reaction which had developed by day 2 after initial infection was characterised by flooding of the alveoli by oedema and neutrophils together with a mild degree of bronchiolar epithelial necrosis. This progressed to an acute exudative fibrinous pneumonia with extensive involvement of the interlobular septa and often with pleurisy. Subsequently, these pulmonary lesions became walled off by fibrous tissue which became infiltrated by plasma cells and lymphocytes. At this stage organisms could be demonstrated only within these nodules in the lung tissue.

Animals↗

Effect of anti-prostaglandin therapy in experimental parainfluenza type 3 pneumonia in weaned, conventional calves.

An acute pneumonia was induced experimentally in 10, 10- to 12-week-old conventional calves by administration into the upper airways of a pathogenic strain of parainfluenza type 3 (PI3) virus. The experimental calves had been selected on the basis of freedom from clinical evidence of respiratory and other diseases, freedom from current infection by PI3 virus as judged by repeated nasopharyngeal swabbing and freedom from earlier PI3 virus infection as judged by their lack of significant levels of serum antibody to that virus. The infection procedure was deemed to have been successful in that infection was established with subsequent seroconversion, clinical signs of a febrile pneumonia arose soon after the administration of virus, histopathological changes characteristic of PI3 pneumonia developed and the presence of PI3 virus antigen was demonstrated by immunofluorescence in association with those lesions. Treatment of five of the pneumonic calves was carried out on days 1, 2 and 3 of the trial using the anti-prostaglandin compound flunixin meglumine and that treatment appeared to be of benefit in that in the test calves there was a prompt cessation of coughing with fewer fevers and lower respiratory rates as compared with the untreated controls. The drug did not appear to influence PI3 infection rates but its administration was associated with a marked reduction in the extent of pulmonary consolidation, probably as the result of its known ability to limit the acute inflammatory response.

Acute Disease↗

Experimental production of bovine pneumonic pasteurellosis.

Pneumonic pasteurellosis has been reproduced in conventional, weaned, Friesian-cross calves using a strain of Pasteurella haemolytica biotype A, serotype 1 (P haemolytica A1) isolated from a pathologically confirmed incident of bovine pneumonic pasteurellosis. The major clinical findings were pyrexia, hyperpnoea, tachypnoea, nasal discharge and reduced appetite. Fibrinous pneumonia was present in the lungs of animals at necropsy on days 2 and 3 after initial infection while by days 9 and 10 after initial infection many of the areas of fibrinous pneumonia were confined by a fibrous capsule forming well defined nodules. During the experiment natural transmission of the infecting strain of P haemolytica A1 occurred in two control calves which developed a condition identical to that in the artificially infected calves. P haemolytica A1 was repeatedly recovered from the nasopharynx of infected calves and at necropsy throughout the upper and lower respiratory tracts. Seroconversion, as measured by indirect haemagglutination, to the organism developed in all infected calves by days 9 and 10 after initial infection. The clinical, microbiological and pathological findings were identical to those seen in field incidents of bovine pneumonic pasteurellosis involving recently housed, weaned, single-suckled calves.

Animals↗

Bovine nasal granuloma (atopic rhinitis) in Britain.

During three successive summer grazing periods, several young Friesian cross heifers developed an increasingly severe upper respiratory syndrome which improved soon after the animals were housed. The major clinical signs during the third summer episode were: ocular and nasal discharge; ulceration of the nasal mucosa, which was swollen, causing obstruction of the nasal passages; and the presence of large numbers of small, hard, white nodules in the nasal vestibules. From these clinical and epidemiological findings, a diagnosis of atopic rhinitis was made.

Animals↗