J Wildl Dis, 1997 Oct, 33(4), 738 - 48 Evaluation of a multivalent Pasteurella haemolytica vaccine in bighorn sheep: safety and serologic responses; Miller MW et al.; We examined effects of a multivalent Pasteurella haemolytica vaccine (serotypes A1, A2, T10) on humoral immune responses and P . haemolytica isolation rates in bighorn sheep (Ovis canadensis) . Thirty captive bighorns, divided into groups of three on the basis of age, sex, and previous history of pneumonic pasteurellosis, received 0, 1, or 2 vaccine doses . Mild, transient lameness in most bighorns 1 day after initial vaccination was the only adverse effect observed . Oropharyngeal (> or = 75%) and nasal (< or = 50%) isolation rates for P . haemolytica did not differ among treatment groups . Ten of 36 distinguishable biogroup variants accounted for about 87% of the 464 P . haemolytica isolates from bighorns, but prevalences of specific biogroups were not affected by vaccination . Bighorns receiving 1 or 2 vaccine doses showed marked elevations in leukotoxin neutralizing antibody titers beginning 1 wk after vaccination . Agglutinating antibody titers to serotype A1 and A2 surface antigens were also elevated in vaccinated bighorns within 2 wk after vaccination; agglutinating antibody titers to serotype T10 surface antigens were relatively high in all three groups but appeared unaffected by vaccination . Vaccination 7 to 14 wk prior to parturition elevated leukotoxin neutralizing antibody titers in colostrum, but neither leukotoxin neutralizing nor serotype A1 surface antigen agglutinating antibody titers differed through 16 wk of age among lambs born to dams from different vaccine dose groups . Our data demonstrate that this multivalent P . haemolytica vaccine is safe and stimulates marked antibody responses in bighorn sheep . Further evaluation of this vaccine as a tool in preventing and managing pasteurellosis in bighorn sheep appears warranted. Vet Clin North Am Food Anim Pract, 1997 Nov, 13(3), 471 - 81 Bovine respiratory tract disease caused by bovine viral diarrhea virus; Potgieter LN; Although several viruses and bacteria are capable of inducing bovine respiratory tract disease, a pivotal organism in the cause of this complex disease may be bovine viral diarrhea virus (BVDV) . Circumstantial evidence has long supported this hypothesis . It is frequently present in diseased respiratory tract tissues often together with other viruses or bacteria . Field observations suggest marked synergism occurs . Researchers have confirmed that, in most instances, the virus itself elicits only a mild respiratory tract disease in susceptible calves, but some strains may be much more pneumo-pathogenic than others . Experimental evidence now supports the hypothesis, that BVDV markedly enhances respiratory tract disease caused by IBRV, BRSV, or Pasteurella haemolytica; that it impairs pulmonary immunity; and that it, by itself, may produce mild respiratory tract disease. Zentralbl Bakteriol, 1997 Oct, 286(3), 333 - 54 Genotypic relationships among strains classified under the (Pasteurella) haemolytica-complex as indicated by ribotyping and multilocus enzyme electrophoresis; Angen O et al.; Two-hundred and one strains classified under the (Pasteurella) haemolytica-complex isolated from cattle, sheep, deer, pigs, hares and rabbits were investigated by ribotyping . Fifty-nine of these strains were selected for further studies using multilocus enzyme electrophoresis (MEE) . A correlation between the clusters identified by ribotyping and MEE was demonstrated and the results furthermore indicated that a genetic basis exists for most clusters previously outlined by the use of quantitative evaluation of phenotypic data . The taxonomic relevance of ornithine decarboxylase and fermentation of L-arabinose, D-sorbitol and glucosides for taxonomic delineation within the (P.) haemolytica-complex was supported . A taxonomic importance was further indicated for ONPG, ONPX, ONPF, meso-inositol, D-xylose, maltose, dextrine and NPG in relation to some of the taxa . Within the porcine taxon 15, however, differences in ornithine decarboxylase did not correspond to genetic clusters . Six lineages were revealed by MEE . Lineage A contained electrophoretic types (ETs) representing biogroups 1, 3A-3H, 8A and 9, indicating a genetic relationship between these groups--an observation which was supported by ribotyping . Lineage B included biogroup 8D, 3 strains from biogroup 10 and a single strain from biogroup 1 and taxon 18/biovar 1 . Lineage C contained strains allocated to biogroup 6 from ruminants and the porcine taxon 15 . The similarity between these two groups was accentuated by ribotyping . Lineage D and the single isolate in lineage E contained strains allocated to biogroups 7, 10, 8B and 8C, in addition to single strains from biogroups 6 and 9 . The same strains were found in the heterogenous ribotype cluster 17 . Lineage F contained strains representing the leprine taxon 20 and the ruminant (P.) granulomatis . Ribotyping indicated that the ruminant biogroup 3J was affiliated with both taxon 20 and (P.) granulomatis. Zentralbl Bakteriol, 1997 Oct, 286(3), 317 - 32 Further studies of the relationships among strains classified as taxon 15, taxon 18, taxon 20, (Pasteurella) granulomatis or the (Pasteurella) haemolytica-complex in ruminants using quantitative evaluation of phenotypic data; Angen O et al.; Ninety-three trehalose-negative (P.) haemolytica-like strains of ruminant, porcine and leprine origin were investigated . A quantitative evaluation of phenotypic tests was used and the results obtained were compared with those from 246 previously investigated ruminant strains . Cluster analysis of the results obtained displayed most of the taxa as distinct groups which could be related to differences in key characters . Although only minor phenotypic differences were observed between the taxa investigated and the taxa were internally heterogeneous for many of the tests, it was possible to identify characters separating most groups . However, in three instances, taxa isolated from different species could not be separated by any of the tests used or by quantitative evaluation of all 79 tests--the only difference being the species of animals from which they had been isolated . Taxa which could not be separated by phenotypic tests included the ruminant biogroup 6 of (P.) haemolytica and the porcine taxon 15/biovar 1, the ruminant biogroup 7 of (P.) haemolytica and the porcine taxon 15/biovar 2, and ruminant biogroup 31 of (P.) haemolytica and the leprine taxon 20/biovar 1. Am J Vet Res, 1997 Nov, 58(11), 1227 - 31 Effects of Pasteurella haemolytica leukotoxin and lipopolysaccharide on histamine, prostanoid, and leukotriene release by bovine lung parenchyma in vitro; Saban R et al.; OBJECTIVE: To identify the effect of Pasteurella haemolytica lipopolysaccharide (LPS) and leukotoxin (LKT) on spontaneous and calcium ionophore-induced histamine and inflammatory mediator release from isolated bovine lung parenchyma . SAMPLE POPULATION: Lungs from 8 healthy cattle . PROCEDURE: Isolated bovine lung parenchyma was incubated in vitro for 2 hours with LKT or LPS, and spontaneous and induced release of inflammatory mediators was determined . RESULTS: LKT and LPS increased spontaneous release of histamine and leukotriene B4 . In addition, incubation with LPS increased spontaneous release of prostaglandin E2 . Moreover, a differential effect of the 2 toxins on calcium ionophore-induced inflammatory mediator release was observed . LKT specifically primed isolated lung parenchyma to release leukotriene B4 and thromboxane B2 in response to calcium ionophore, whereas LPS did not alter the profile of prostanoids released by bovine lung tissue exposed to calcium ionophore . CONCLUSIONS: Pasteurella haemolytica toxins have a direct effect on bovine lung parenchyma, causing release of inflammatory mediators, which contribute to response to infection . Furthermore, bacterial toxins (LKT in this study) may sensitize tissues to the effects of other irritant stimuli, amplifying the inflammatory response. J Wildl Dis, 1996 Oct, 32(4), 594 - 602 Experimental contact of bighorn sheep (Ovis canadensis) with horses and cattle, and comparison of neutrophil sensitivity to Pasteurella haemolytica cytotoxins; Foreyt WJ et al.; Peripheral blood neutrophils from horses, cattle, and Rocky Mountain bighorn sheep (Ovis canadensis canadensis) were evaluated for susceptibility to cytotoxin-dependent lysis of different biotypes and serotypes of Pasteurella haemolytica of domestic sheep, cattle, bighorn sheep, or mountain goat (Oreamnos americana) origin utilizing a cytotoxicity assay which measures the degree of bacteria cytotoxin-killing of neutrophils . All isolates of P . haemolytica (biotypes A and T) were noncytotoxic to horse neutrophils . Thirteen of 18 R haemolytica biotype A isolates were cytotoxic (> 50% neutrophil death in vitro) to bighorn sheep neutrophils, and four of 10 P . haemolytica biotype A isolates were cytotoxic to neutrophils of cattle; P . haemolytica biotype T (= Pasteurella trehelosi) isolates were noncytotoxic to neutrophils of bighorn sheep and cattle . When six bighorn sheep were pastured with three horses, only P . haemolytica biotype T isolates were recovered from the bighorn sheep throughout the study; Pasteurella spp . organisms were not isolated from the three horses . At initiation of a study where five bighorn sheep were pastured with three cattle, P . haemolytica biotype A, serotype 1, 2 was isolated from all three cattle, and only P . haemolytica biotype T isolates were recovered from the bighorn sheep . One bighorn sheep died in each of the horse and cattle copasturing experiments . Pasteurella haemolytica was not isolated from the bighorn sheep which died in the horse copasturing experiment . A noncytotoxic P . haemolytica biotype A, serotype 2 was isolated at necropsy from the bighorn which died in the cattle contact experiment . Based on these experiments, we believe bighorn sheep and horse association would not be detrimental to bighorns due to P . haemolytica induced pneumonia. J Wildl Dis, 1996 Oct, 32(4), 586 - 93 Susceptibility of Dall sheep (Ovis dalli dalli) to pneumonia caused by Pasteurella haemolytica; Foreyt WJ et al.; We evaluated susceptibility of Dall sheep (Ovis dalli dalli) to bacterial pneumonia induced by two strains of Pasteurella haemolytica of domestic sheep origin by evaluating the sensitivity of blood neutrophils of eight Dall sheep to lysis by cytotoxins of P . haemolytica, and by intratracheal inoculation of three Dall sheep, two bighorn sheep (Ovis canadensis), and two domestic sheep with 3.7 x 10(6) or 2.5 x 10(7) colony forming units of P . haemolytica . Neutrophils from the Dall sheep were more sensitive to lysis by cytotoxins from supernatants of a P . haemolytica, biotype A, serotype 2 (A2), of domestic sheep origin, than were neutrophils from six bighorn sheep . This cytotoxic bacterium was the same isolate that was used for intratracheal inoculation of two Dall sheep and two domestic sheep . Inoculation of this cytotoxic P . haemolytica A2 resulted in fatal fibrinopurulent pleuropneumonia in the first Dall sheep within 24 hr of inoculation, and pneumonic lesions in the second Dall sheep before it was euthanized 52 hr after inoculation . This strain of P . haemolytica A2 did not cause respiratory disease when inoculated into two domestic sheep . A noncytotoxic strain of P . haemolytica; biotype T, serotype 3,4,10 of domestic sheep origin did not result in pneumonia in the third Dall sheep or two bighorn sheep . Prior to inoculation, P . haemolytica, biotype T isolates were obtained from all three Dall sheep, but none of these isolates was cytotoxic . At necropsy, cytotoxic P . haemolytica A2 was isolated from lungs and other tissues of the two pneumonic Dall sheep . Based on these results, we conclude that Dall sheep appear to be at least as sensitive as bighorn sheep to pneumonia caused by P . haemolytica A2 of domestic sheep origin . Because in vitro and in vivo results appear closely correlated in this and other studies, we believe with additional evaluation and standardization, neutrophil cytotoxicity tests may serve as a substitute for live animal challenges in future studies of pathogenic P . haemolytica in wild sheep. Avian Dis, 1997 Jul-Sep, 41(3), 676 - 82 Detection of Pasteurella multocida-specific DNA in turkey flocks by use of the polymerase chain reaction; Kasten RW et al.; A polymerase chain reaction (PCR)-based assay using primers constructed to amplify the gene (psl) encoding the P6-like protein (Psl) of Pasteurella multocida was developed . After Southern blotting and hybridization with psl, the assay (PCR-H) was found to be specific (it did not detect a variety of other avian bacterial pathogens) and sensitive (detected > or = 10 P . multocida organisms or > or = 24 femtograms of extracted P . multocida DNA) . Samples were collected from the oropharynx of randomly selected birds housed on premises that had recently experienced an outbreak of avian cholera (outbreak farms) or from birds housed on premises that had not reported an outbreak of this disease during the preceding 12 mo (control farms) . The PCR-H assay detected 11 infected turkeys out of a total of 178 sampled on six outbreak farms as compared with isolation of P . multocida from 23 turkeys by using mouse inoculation . Neither method detected P . multocida in samples collected from 174 turkeys sampled on six control farms . Statistical analysis using the Kappa test demonstrated that the results of the two tests showed poor agreement from five outbreak flocks (K = 0, 0, 0, 0.35, 0.47) and strong agreement from one outbreak flock (K = 0.89) . Combined results from the outbreak flocks showed poor agreement (K = 0.49) between the two methods. Infect Immun, 1997 Nov, 65(11), 4502 - 8 Role of outer membrane protein H (OmpH)- and OmpA-specific monoclonal antibodies from hybridoma tumors in protection of mice against Pasteurella multocida; Vasfi Marandi M et al.; Two major outer membrane proteins of Pasteurella multocida, designated OmpH and OmpA, were characterized and shown to be related to the families of porin and heat-modifiable proteins, respectively . The backpack hybridoma tumor system in BALB/c mice was used to continuously deliver immunoglobulin G2b (IgG2b) monoclonal antibodies (MAbs) specific for OmpH (MAb MT1) and OmpA (MAb MT4.1) . MAbs were detected in serum and peritoneal lavage samples of mice bearing hybridoma tumors by an enzyme-linked immunosorbent assay and an immunoblot assay . Highly significant protection was observed in mice bearing MT1 hybridoma tumors against both intraperitoneal and intranasal challenge infections with homologous nontoxigenic P . multocida strains possessing MAb MT1-reacting epitopes, whereas the mice bearing MT4.1 hybridoma tumors were not protected . The numbers of P . multocida organisms in the lungs of mice bearing MT1 hybridoma tumors were significantly less than those in lungs of mice bearing MT4.1 hybridoma tumors at 48 h postchallenge . These results indicate that the OmpH-specific MAb inhibited proliferation of P . multocida in the lungs . MAb MT1 was unable to kill P . multocida in vitro in the presence of complement . However, an enhanced phagocytosis by polymorphonuclear cells (PMNs) was observed in mice bearing MT1 hybridoma tumors . P . multocida induced a more extensive and rapid influx of PMNs into the peritoneal cavity of mice bearing MT1 hybridoma tumors than of mice bearing MT4.1 hybridoma tumors . The results of this study demonstrate for the first time that IgG MAbs against OmpH of P . multocida are involved in the protection of mice against lethal challenge infection by means of opsonization and inhibition of proliferation of P . multocida as a result of increased influx of PMNs into the infection site. J Comp Pathol, 1997 Aug, 117(2), 137 - 45 Influence of immunization on the pulmonary inflammatory response of rabbits induced by Pasteurella haemolytica A1 lipopolysaccharide; Ramirez-Romero R et al.; Immune complex formation has long been thought to play a role in the pathogenesis of Pasteurella haemolytica pneumonia . This study in laboratory rabbits was designed to investigate immune-mediated damage in respiratory tissue caused by lipopolysaccharide (LPS) . Severe lesions were induced by the intratracheal (IT) injection of P . haemolytica A1 LPS (50 micrograms) into rabbits previously immunized with P . haemolytica killed whole cells emulsified with Freund's incomplete adjuvant (FIA); these lesions included perivascular oedema and polymorphonuclear leucocyte (PMN) infiltration of the subintima, with degeneration and necrosis of the media . Smaller vessels were occluded by PMNs in various stages of degranulation . PMN counts in bronchoalveolar lavage (BAL) fluid were significantly elevated (P < 0.05) . Lesions were also induced by the IT injection of LPS (50 micrograms) into rabbits pretreated with an emulsion consisting merely of FIA and formol-saline; these lesions included moderate to severe congestion, interstitial oedema, alveolar serofibrinous exudation and PMN infiltration . PMNs were also present in BAL fluid . Rabbits pretreated with FIA in formol-saline and given a later IT injection of saline, and rabbits pretreated with bovine serum albumin (BSA) in FIA and given a later IT injection of BSA, were included as negative and positive control groups . Cutaneous lesions were also induced by the intradermal injection of LPS into rabbits immunized against P . haemolytica and of BSA into rabbits immunized with BSA . Overall, the pulmonary and cutaneous lesions induced in vaccinated rabbits by antigen administration were more severe than those seen in non-vaccinated rabbits . The lesions in rabbits, which were similar to those seen in natural cases of P . haemolytica pneumonia in cattle, were characterized by a fibrinopurulent inflammatory process with extensive interstitial oedema, fibrinous exudate, and PMNs . This model may help to elucidate the pathogenesis of pneumonic pasteurellosis in immunized animals. FEMS Microbiol Lett, 1997 Oct 15, 155(2), 203 - 7 Protection against haemorrhagic septicaemia induced by vaccination of buffalo calves with an improved oil adjuvant vaccine; Shah NH et al.; An experimental oil adjuvant vaccine was developed against haemorrhagic septicaemia, a disease of cattle and buffalo caused by Pasteurella multocida serotype B and E . Mineral oil, Mercol 52, was used as adjuvant together with Span 85 and Tween 85 as emulsifiers . The vaccine was evaluated by single dose intramuscular immunisation of 1-2 year old buffalo calves . IgG and IgM class antibodies were determined by ELISA . The group of animals immunised with the experimental oil adjuvant vaccine showed a high titre of the IgG class of antibodies measured at 300 days post vaccination . To compare the protective efficacy of the vaccine with the commonly used broth bacterin, another group of buffalo calves was immunised by broth bacterin . This group showed a low level of IgG antibodies . Protection was assessed by challenge with 10(9) viable bacteria of P . multocida type B:2,5 administered subcutaneously, 250 days post vaccination . Animals vaccinated with the experimental oil adjuvant vaccine were fully protected . The other groups of animals, vaccinated with broth bacterin or used as control (non-vaccinated), developed symptoms of haemorrhagic septicaemia . A strong relationship between IgG but not IgM class antibody level and resistance to challenge was observed . The experiment demonstrated that the experimental oil adjuvant vaccine was superior to broth bacterin in providing protection against experimental haemorrhagic septicaemia in young buffalo calves beyond 250 days. Vet Pathol, 1997 Sep, 34(5), 421 - 30 Effects of Pasteurella multocida toxin on porcine bone marrow cell differentiation into osteoclasts and osteoblasts; Gwaltney SM et al.; The effect of Pasteurella multocida toxin (PMT) on porcine osteoclast and osteoblast differentiation was studied using in vitro cell culture systems . When grown in the presence of Vitamin D3, isolated porcine bone marrow cells formed multinucleated cells with features characteristic of osteoclasts . Exposure of bone marrow cells to Vitamin D3 and PMT during growth resulted in formation of increased numbers and earlier appearance of osteoclasts compared to controls . Ultrafiltered medium form PMT-treated cells likewise increased osteoclast numbers, suggesting that a soluble mediator may be involved in the action of PMT . When cell cultures were treated with fluorescein-labeled PMT, fluorescence was found within the cytoplasm of small, round cells that did not resemble either osteoclasts or osteoclastic precursor cells . Cultures of porcine bone marrow cells exposed to dexamethasone, ascorbic acid, and beta-glycerophosphate developed into osteoblastic cells that formed multilayered, mineralized nodules . Exposure of osteoblastic cultures to low concentration of PMT resulted in retarded cell growth, formation of decreased numbers of nodules and minimal to no mineralization in the nodules; higher concentration of PMT resulted in increased cellular debris and poor growth of cells, with no nodule formation . These findings suggest that PMT may induce turbinate atrophy in pigs by increasing osteoclast numbers and inhibiting osteoblastic bone formation . The effect of PMT on osteoclastic differentiation and growth may not be due to a direct effect on preosteoclastic cells, but rather due to alterations in the soluble mediator secretion by marrow stromal cells. Am J Pathol, 1997 Oct, 151(4), 1009 - 17 Respiratory infection in lipid-fed rabbits enhances sudanophilia and the expression of VCAM-1; Richardson M et al.; The pathogenesis of atherosclerosis has been related to infection of the arterial wall, but it is not clear whether this occurs before or after the development of lipid-containing lesions . Respiratory bacterial infection increases the expression of vascular cell adhesion molecule-1 (VCAM-1) . We therefore examined whether a similar infection would enhance atherosclerosis in New Zealand White rabbits fed chow supplemented by 15% (w/w) egg yolk for 50 days . Rabbits with naturally acquired respiratory infection by Pasteurella multocida, pathogen-free (SPF) animals infected by P . multocida in the laboratory, and age-matched SPF rabbits maintained in a disease-free environment were used . Endothelial cells expressing VCAM-1 in the aorta between intercostal arteries 3 and 5 were identified using anti-VCAM-1 (Rb1/9) and an alkaline-phosphatase-linked secondary antibody and quantified in Hautchen preparations . The remainder of the aorta was stained with Sudan IV to show lipid deposition . The expression of VCAM-1 (mean +/- SEM per 10,000 cells) was 22 +/- 8 (n = 5) in the lipid-fed SPF rabbits, significantly different from that in the lipid-fed rabbits with naturally occurring infection (190 +/- 51 (n = 5)) or from rabbits infected in the laboratory (106 +/- 25 (n = 5)) . The extent of Sudanophilia was significantly greater in the naturally infected rabbits (8.3 +/- 1.2%) or infected SPF rabbits (10.3 +/- 1.8%) than in the SPF rabbits (2.7 +/- 0.8%; P < 0.05) . Antibiotic treatment in naturally infected rabbits reduced the number of cells expressing VCAM-1 and the extent of the Sudanophilia to baseline levels . Thus, Sudanophilia is enhanced by bacterial infection in rabbits fed egg yolk and is associated with a significant increase in VCAM-1. Equine Vet J, 1997 Sep, 29(5), 394 - 9 Bacterial endocarditis in horses: ten cases (1984-1995); Maxson AD et al.; A retrospective study of 10 horses with bacterial endocarditis was performed in order to describe the echocardiographic findings in horses with bacterial endocarditis, in conjunction with clinical signs and post mortem findings, and to evaluate the usefulness and the formulation of a prognosis . Echocardiographic and post mortem examinations were performed in 7 horses . Post mortem examination alone was performed in 2 horses and echocardiographic examination alone performed in one horse . No breed or sex predilection was obvious . Mean age +/- s.d . was 2.12 +/- 3.32 years . Predominant clinical signs and abnormal clinical pathology data were fever, cardiac murmur, tachycardia, tachypnoea, hyperfibrinogenaemia, anaemia and leucocytosis . Pasteurella/Actinobacillus spp . and Streptococcus spp . were most commonly cultured . Vegetative lesions were found most frequently on the mitral valve and secondarily on the aortic valve . The location and number of lesions identified with echocardiography in the horses accurately described the lesions found on post mortem examination . Medical treatment was attempted in 50% of the horses . Serial echocardiography was used to assess the response to treatment in 2 horses . All horses with vegetative lesions of the mitral and/or aortic valve died or were subjected to euthanasia due to the severity of their cardiac disease . Both horses with tricuspid valve endocarditis were cured of the infection, one horse returned to racing after antimicrobial therapy and the other was subjected to euthanasia due to severe laminitis. Curr Microbiol, 1997 Nov, 35(5), 316 - 8 Isolation of Pasteurella haemolytica from grass, drinking water, and straw bedding used by sheep; Burriel AR; Pasteurella haemolytica was isolated from three of 18 grass samples and four of 18 water samples collected from two grazing fields occupied by sheep . This microorganism was also isolated from three of nine straw bedding samples collected from a pen housing ewes affected by mastitis caused by P . haemolytica . The same ewes developed scabbed papilloma-like lesions on the teat and udder skin . These lesions were colonized by P . haemolytica of various serotypes . Colder, wetter weather seems to prolong the survival of P . haemolytica in the environment of sheep . Survival of virulent strains of P . haemolytica in the environment could accumulatively increase the bacterial count, contributing to their transmission from animal to animal . The preference of P . haemolytica for colder, wetter conditions was confirmed in the laboratory where this microorganism survived longer in distilled water, phosphate-buffered saline, Todd-Hewitt broth, and ewe's milk kept at 4 degrees C. Dtsch Tierarztl Wochenschr, 1996 Dec, 103(12), 513 - 6 {Importance of blood serological examinations in the context of rhinitis atrophicans diagnosis}; Muller E et al.; In a period of three years in 95 breeding-herds, which were free from Rhinitis atrophicans (R . a.), 5001 blood-samples were examined . All samples were examined by the SNT/EBL-cell-culture-test-mostly however by the SNT/ELISA-system- and were free of antibodies against R . a . On the contrary in 6 herds, that had bought swines from latent infected populations, antibodies against the toxin could be found before clinical signs were to be seen and before toxin producing pasteurellas could be discovered, too . In other 4 herds antibodies against R . a . could be found . In the last mentioned herds R . a . was suspected by clinical, bacteriological and pathomorphological examinations . The serological determination of blood may replace the provement of the diameter of the nose by pathomorphological diagnosis . The serological investigation is a good test . After positive results of antibodies comprehensive bacteriological tests should follow . This can be integrated in the diagnostical system of R . a . very easily, because enough blood samples will be taken ordered by the official examinations for European Swinefever- and Aujeszky-disease. Cell Stress Chaperones, 1997 Sep, 2(3), 180 - 90 Refolding of recombinant Pasteurella haemolytica A1 glycoprotease expressed in an Escherichia coli thioredoxin gene fusion system; Watt MA et al.; Pasteurella haemolytica A1 secretes an O-sialoglycoprotein endopeptidase (EC . 3.4.24.57) (glycoprotease: Gcp) which is specific for O-linked sialoglycoproteins . When the cloned gene is expressed in Escherichia coli, the recombinant glycoprotease (rGcp) is secreted to the periplasm where it is present as a disulfide-linked aggregate which lacks enzymatic activity . In vitro refolding and activation of rGcp by mammalian protein disulfide isomerase (PDI) or by the E . coli chaperones (DnaK, DnaJ and GrpE) indicate that the redox environment of rGcp is critical in restoring biological activity . A fusion protein, rTrx-Gcp, was constructed to investigate the role of thioredoxin (E . coli TrxA) in the production of enzymatically active rGcp . This 47 kDa protein was expressed at a high level, in a soluble, monomeric form, in the cytoplasm of E . coli . Cleavage of the fusion protein by enterokinase released the rGcp fragment (35 kDa) with glycoprotease activity . A higher recombinant glycoprotease activity was recovered after anion exchange chromatography of lysates of E . coli expressing rTrx-Gcp . Thus when E . coli TrxA is combined in a recombinant fusion protein with P . haemolytica A1 Gcp, productive folding of the glycoprotease can occur as a result of the chaperone action of the protein disulfide reductase coupled with its ability to retain the fusion gene product in the E . coli cytoplasm. Zh Mikrobiol Epidemiol Immunobiol, 1997 May-Jun, (3), 15 - 9 {The cultural and morphological properties of different serovariants of Pasteurella multocida when cultured in nutrient broths of different compositions}; Popova TE; The results of the study of the cultural morphological properties and growth dynamics of P . multocida, serotypes A, B, D, in the course of cultivation in liquid media are presented . The bacteria were grown in synthetic nutrient medium 199 and Hottinger broth . The influence of glucose at different concentrations on the growth activity of P . multocida in the synthetic medium was established . The process of the accumulation of surface (capsular) antigens in culture fluid was studied . The levels of the titers of surface antigens were shown to depend on the growth phases of bacteria. Vaccine, 1997 Aug-Sep, 15(12-13), 1423 - 9 Serum antibody responses of cattle vaccinated with partially purified native Pasteurella haemolytica leukotoxin; Confer AW et al.; The objective of these experiments was to study the serum antibody responses of cattle to partially purified, native Pasteurella haemolytica A1 leukotoxin (LKT) formulated with a commercial aluminum hydroxide-DDA-bromide adjuvant . In two experiments, calves received two intramuscular injections 21 days apart and sera were obtained periodically . Serum antibody responses to P . haemolytica outer membrane proteins (OMPs), formalinized P . haemolytica, and LKT were determined . In Experiment A, Holstein calves (140 kg each) were vaccinated with either 10, 1.0 or 0.1 micrograms of LKT, 10(9) c.f.u . of live P . haemolytica, or adjuvanted diluent . In Experiment B, mixed-breed beef calves (200 kg each) were vaccinated with either 100, 50 or 10 micrograms of LKT, 10(9) c.f.u . live P . haemolytica, or adjuvanted diluent . Vaccination of dairy calves with 10 micrograms of partially purified LKT stimulated LKT neutralizing antibody responses similar to those stimulated by vaccination of one calf with live P . haemolytica . In Experiment B, which used larger and different breeds of cattle, two vaccinations 3 weeks apart with 50 micrograms LKT stimulated LKT neutralizing responses equivalent to or greater than those stimulated by vaccination with live P . haemolytica . In both experiments, LKT vaccines stimulated only low antibody responses to formalinized P . haemolytica or to OMPs. J S Afr Vet Assoc, 1997 Jun, 68(2), 55 - 8 Efficacy of doxycycline in a goat model of Pasteurella pneumonia; Ole-Mapenay IM et al.; The clinical efficacy of doxycycline (Doxycen, Cenavisa, Spain), a long-acting preparation, was evaluated for treatment of Pasteurella haemolytica infection in 6 goats . One goat was not infected and served as a control . The disease was induced by intratracheal inoculation of 10(7) to 10(9) cfu of P . haemolytica . Confirmation of respiratory disease was based on evidence of appropriate clinical signs . Before and after initiation of doxycycline treatment on day 10, each goat was examined daily . Three clinical responses to doxycycline treatment were noted . Mean rectal temperatures decreased from 40.1 degrees C to normal, while mean respiratory rate decreased from the pre-treatment value of 32 to 27/min after 4 days . Other clinical signs associated with pneumonia resolved within 3-5 days post treatment . In addition the minimum inhibitory concentration of DOTC for the P . haemolytica isolate was found to be < 0.5 microgram/ml . The present study indicates that DOTC may be a useful antimicrobial agent in the treatment of caprine pasteurellosis. Infect Immun, 1997 Sep, 65(9), 3970 - 5 A putative leucine zipper activator of Pasteurella haemolytica leukotoxin transcription and the potential for modulation of its synthesis by slipped-strand mispairing; Highlander SK et al.; A Pasteurella haemolytica cosmid clone that activates leukotoxin transcription in Escherichia coli has been isolated . The activator locus, alxA, is part of a continuous open reading frame that includes the type I hsdM methylase gene . AlxA and HsdM peptides are processed from a precursor, and translation of the polyprotein can be modulated by slipped-strand mispairing across a pentanucleotide repeat, ACAGC, within the 5' end of alxA-hsdM . Extracts containing AlxA can bind to a leukotoxin promoter fragment. Infect Immun, 1997 Sep, 65(9), 3719 - 24 Binding of Pasteurella haemolytica leukotoxin to bovine leukocytes; Brown JF et al.; Pasteurella haemolytica is the principal bacterial pathogen in the bovine respiratory disease complex . This organism produces an exotoxin (referred to as leukotoxin) during logarithmic-phase growth that is a potent leukocyte-modulating agent . At low concentrations, it activates neutrophils and mononuclear phagocytes to release inflammatory mediators, while at the same time making these cells destined to undergo apoptotic cell death . At higher concentrations, the toxin causes rapid swelling and loss of cell viability . In this study, we demonstrated that toxin binding can be directly evaluated by flow cytometry with biologically active biotinylated leukotoxin . Leukotoxin binding was blocked by the addition of a neutralizing anti-leukotoxin monoclonal antibody and was not detected when bovine leukocytes were incubated with culture filtrates from a mutant strain of P . haemolytica that does not produce biologically active leukotoxin . In addition, treatment of bovine leukocytes with protease K eliminated subsequent binding of leukotoxin, suggesting that there is a protein on the leukocyte surface that is either a leukotoxin binding site or is required for stabilization of leukotoxin binding . We did not detect binding of biotinylated leukotoxin to porcine or human leukocytes, which have been reported previously to be resistant to the lytic effects of the leukotoxin . These findings suggest that there may be a specific binding site for P . haemolytica leukotoxin on bovine but not on porcine or human leukocytes and that it might be involved in the activation and lytic activities of the leukotoxin. Infect Immun, 1997 Sep, 65(9), 3585 - 93 Evolutionary genetics of Pasteurella haemolytica isolates recovered from cattle and sheep; Davies RL et al.; Genetic diversity and relationships among 194 Pasteurella haemolytica isolates, which were recovered predominantly from cattle (39%) and sheep (58%) suffering from pneumonic pasteurellosis in the United Kingdom, Germany, and the United States, were estimated by examination of allelic variation at 18 enzyme-encoding loci detected by multilocus enzyme electrophoresis . The isolates formed two major divisions . One included 178 Pasteurella haemolytica sensu stricto strains representing serotypes A1, A2, A5 to A9, A12 to A14, and A16; the other was composed of 16 isolates belonging to the A11 taxon . P . haemolytica isolates were classified into 22 electrophoretic types (ETs) that formed three primary phylogenetic lineages . One lineage was represented by ovine serotype A2 isolates, a second lineage consisted of bovine serotype A2, together with serotype A7 and A13 isolates, and the third lineage included isolates representing all of the other serotypes, as well as a second group of serotype A7 strains . Electrophoretic types were nonrandomly associated with specific capsular serotypes, lipopolysaccharide (LPS) types, outer membrane protein (OMP) types, and host species . Bovine isolates were represented by only three serotypes (A1, A2, and A6) in 5 ETs, whereas ovine isolates were represented by all of the serotypes in 19 ETs . The majority (76%) of bovine isolates were of serotypes A1 or A6 and belonged to a single ET that marked a virulent, cattle-specific clonal group . Among the ovine isolates, 40% were of serotype A2 and belonged to two ETs that represented two virulent, sheep-specific clonal groups . Bovine A1 and A6 isolates and bovine A2 isolates were phylogenetically distinct from ovine isolates of the same serotypes, indicating that different subpopulations of these serotypes are associated with disease in cattle and sheep . Consistent differences in the OMP profiles of strains of the bovine and ovine lineages of these three serotypes suggest that certain OMPs are involved in host specificity and virulence . Evolutionary relationships among P . haemolytica isolates indicate that the ancestral host is the sheep and that several distinct clonal lineages have crossed the species barrier into cattle . The A11 taxon is a heterogeneous group of opportunistic pathogens of sheep that represents a separate species. Poult Sci, 1997 Sep, 76(9), 1248 - 55 Application of a nonlinear regression function to evaluate the kinetics of antibody response to vaccines in chicken lines divergently selected for multitrait immune response; Weigend S et al.; To evaluate the kinetics of immune response to vaccines in chickens, antibody response curves were approximated to the observed antibody ratios by using a nonlinear regression function . New parameters, the curve maximum (ymax) and the time of the maximum (tmax), were calculated . The method was applied to analyze the kinetics of the serum antibody response to Mycoplasma gallisepticum (MG) and Pasteurella multocida (PM) vaccines in White Leghorn lines selected, in replicate, for 10 generations for high (High) and low (Low) multitrait immune response . Chicks were immunized at 6 wk of age with both vaccines . Serum antibody levels were analyzed before (0) and 1, 2, 3, 4, 5, 12, and 21 wk postvaccination (wpv) . The High lines displayed a significantly higher response than Low to both MG and PM . The difference in ymax between High and Low lines was 3.25-fold for PM response and 1.5-fold for MG response . Low lines had a significantly (P < 0.05) later tmax than High lines to MG, but not to PM . There was a significant (P < 0.05) positive correlation between the antibody responses to MG and PM, in High lines for the antibody ratios 0, 3, and 21 wpv and in Low lines for 0, 12, and 21 wpv . The ymax and tmax of antibody responses to the two vaccines were not correlated . The results on the kinetic differences of the antibody responses to MG and PM suggest that the kinetics and persistence of antibody reaction have different genetic regulation in response to each vaccine. Proc Natl Acad Sci U S A, 1997 Aug 5, 94(16), 8686 - 90 Epithelial antibiotic induced in states of disease; Stolzenberg ED et al.; Epithelial defensins provide an active defense against the external microbial environment . We investigated the distribution and expression of this class of antimicrobial peptides in normal cattle and in animals in varying states of disease . beta-defensin mRNA was found to be widely expressed in numerous exposed epithelia but was found at higher levels in tissues that are constantly exposed to and colonized by microorganisms . We observed induction in ileal mucosa during chronic infection with Mycobacterium paratuberculosis and in bronchial epithelium after acute infection with Pasteurella haemolytica . It has been proposed that expression of antimicrobial peptides is an integral component of the inflammatory response . The results reported here support this hypothesis and suggest that epithelial defensins provide a rapidly mobilized local defense against infectious organisms. Vaccine, 1997 Aug, 15(11), 1254 - 60 Protection, humoral and cell-mediated immune responses in calves immunized with multiple emulsion haemorrhagic septicaemia vaccine; Verma R et al.; A multiple emulsion (ME), vaccine against Pasteurella multocida (P52) infection in cattle was prepared and the efficiency in terms of immunity to direct challenge, duration of this immunity for up to 1 year and the role of humoral and cell-mediated immune mechanisms were studied . ME vaccine was sterile, safe and was potent when tested in rabbits and calves . Nineteen calves were immunized with a single 4 ml dose of ME vaccine intramuscularly . Group of these calves were challenge infected with virulent P . multocida (P52) (10(-1) 18 h old broth culture) given by the subcutaneous route at 21 days, 3 months, 6 months, 9 months and 1 year . All the immunized calves withstood challenge infection and showed 100% protection . Humoral immune response was measured by indirect haemagglutination test (IHA) and enzyme-linked immunosorbant assay (ELISA) . Statistically ELISA values were found to be superior to IHA values because of small coefficient of variance . A fall in mean antibody titres during 24 h, 48 h, post-challenge infection was recorded whereas a steady increase in the titre after 72 h up to 10 days was noticed . The prechallenge mean titre in animals correlated with survival of animals . Humoral antibodies were detected as early as 7 day post-immunization and persisted to 1 year after immunization . Leucocyte migration inhibition test showed > 20% migration inhibition during all pre- and post-challenge periods in animals suggesting an involvement of cell-mediated immune mechanism in protection . Our findings suggested that both humoral and cell-mediated immune responses contribute to protection in vaccinated animals . The results of these studies of ME vaccine showed that it can be successfully used for the effective control of haemorrhagic septicaemia. Microbiology, 1997 Aug, 143 ( Pt 8), 2841 - 9 Genetic relationships among Pasteurella trehalosi isolates based on multilocus enzyme electrophoresis; Davies RL et al.; Genetic diversity among 60 British Pasteurella trehalosi isolates representing the four recognized capsular serotypes, T3, T4, T10 and T15, and recovered predominantly from sheep suffering from systemic pasteurellosis, was estimated by analysing electrophoretically demonstrable allelic variation at structural genes encoding 19 enzymes . Thirteen of the locl were polymorphic and 20 distinctive multilocus genotypes (electrophoretic types, ETs) were identified . The population structure of P . trehalosi is clonal and its genetic diversity is limited compared with most other pathogenic bacteria . ETs represent clones, and isolates of the same ET were generally associated with the same combination of serotype, LPS type and outer-membrane protein (OMP) type . The genetic diversity of isolates within each of the capsular serotypes varied . Serotype T10 was represented by 18 isolates in two related ETs and exhibited little diversity . By contrast, serotype T15 was represented by 18 isolates in nine ETs and was almost as diverse as the species as a whole Serotype T4 was represented by 18 isolates in five ETs and was less diverse than serotype T15 . Although serotype T3 was more diverse than serotype T15 it was represented by only three isolates . With the exception of the T10 isolates and those recovered from healthy sheep, 35 disease isolates belonged to 16 ETs, each of which was represented by only one to four isolates . The fact that a high proportion of disease is caused by a relatively large number of clones suggests that P . trehalosi is essentially an opportunistic pathogen . In addition to having the same capsular structure, isolates belonging to the two T10 clones were characterized by possession of similar, if not identical, O-antigens (LPS types 2 and 4) . The occurrence of 18 serotype T10 isolates in only two ETs suggests that the T10 capsule and type 2/4 O-antigen confer enhanced virulence on members of these two clones . Multilocus enzyme electrophoresis (MLEE) had greater resolving power than did capsule/LPS/OMP analysis, being able to distinguish 20 rather than 14 sub-divisions within P . trehalosi . The technique demonstrated genetic identity or non-identity among strains of the same or different serotypes from different geographic localities within the UK and was a useful epidemiological tool. Vet Res Commun, 1997 Aug, 21(6), 453 - 62 Aspects of the pharmacokinetics of doxycycline given to healthy and pneumonic East African dwarf goats by intramuscular injection; Ole-Mapenay IM et al.; The effect of experimentally induced Pasteurella haemolytica pneumonia on the pharmacokinetics of doxycycline (Doxycen Retard) administered intramuscularly was studied in seven East African dwarf goats . The study was conducted in two consecutive phases, separated by a washout period of four weeks . The experimental infection, induced by intratracheal administration of 5 ml of 10(7) to 10(9) cfu/ml of Pasteurella haemolytica, produced a temperature rise, depression and laboured breathing within 6-12 days after inoculation . The concentrations of doxycycline in the serum were determined by a quantitative microbiological assay using an agar-gel diffusion method employing Bacillus cereus var mycoides (ATCC 11778) as the test organism, with a level of detectability of approximately 0.05 micrograms/ml . The concentration-time curve of doxycycline in the serum after intramuscular injection of 20 mg/kg bodyweight of the long-acting formulation before and after experimental infection was adequately described by a one-compartment open model . The maximum serum concentrations (Cmax) of doxycycline were lower in pneumonic goats than in healthy goats (3.87 +/- 0.52 and 5.56 +/- 0.213 micrograms/ml, respectively), suggesting an increased distribution volume in the peripheral compartment . The mean +/- SEM absorption rate (ka) before infection (1.13 +/- 0.02 h-1) was smaller than the after infection (8.23 +/- 3.81 h-1), but the difference was not significant . The apparent elimination half-life (t 1/2 beta) (24.51 +/- 0.02 h) after infection was significantly increased (p < 0.05), while the corresponding rate constant (beta) was decreased (p < 0.01) . The absorption half-life (t 1/2(alpha)) (0.137 +/- 0.03 h) was significantly decreased (p < 0.01) after infection . The distribution volume (Vd(beta)) was significantly increased after infection (p < 0.05) . It is concluded that, although experimental infection had an effect on the disposition kinetics of doxycycline, this was not sufficiently pronounced to require alteration of the dosage during disease. Vet Res Commun, 1997 Aug, 21(6), 421 - 30 Bacterial identity and characteristics in healthy and unhealthy respiratory tracts of sheep and calves; Barbour EK et al.; The aim of this study was to compare different bacteriological aspects of the respiratory systems of healthy (H) versus unhealthy (UH) animals with respiratory signs . The prevalence of different bacterial species was determined in the upper and lower respiratory tract of H and UH Najdi sheep, Somali sheep and Holstein calves . The characteristics of Pasteurella spp . isolates, and the biotype of Pasteurella haemolytica were identified in H and UH animals, Eighteen out of 28 (64.3%) of the identified bacterial species in the upper respiratory tract were more prevalent in the nasal cavities of UH Najdi and Somali sheep and Holstein calves with respiratory signs than in apparently healthy animals; four of the most prevalent bacteria in the upper respiratory system of UH sheep were Moraxella spp., Pseudomonas pseudomallei, Erysipelothrix spp., Pasteurella multocida, while three of the most prevalent bacteria in UH calves were Pasteurella haemolytica, Actinomyces spp., and Pseudomonas aeruginosa . The prevalence of six different bacterial species was greater in the lungs of UH animals, namely Actinomyces pyogenes, Erysipelothrix spp., P . haemolytica, Pasteurella ureae, Staphylococcus aureus, and Staphylococcus epidermidis, which could be risk factors in the complexity of the prevalent respiratory diseases of the animals surveyed . Of the biochemical, cytological and colonial characteristics studied in the identified P . haemolytica and P . multocida, two characters were significantly different (p < 0.05) in organisms isolated from UH as compared to those from H animals . These were the higher loss of haemolytic power by the strains of P . haemolytica and the decreased fermentation of trehalose by all the strains of P . multocida recovered from healthy animals . The only biotype of P . haemolytica isolated from H animals was biotype A, while both biotypes A (88.0% of the isolates) and T (12.0% of the isolates) were recovered from UH animals. J Rheumatol, 1997 Aug, 24(8), 1649 - 52 Pasteurella multocida infectious arthritis with acute gout after a cat bite; Butt TS et al.; A 74-year-old man with chronic lymphocytic leukemia, immune purpura, and gout presented with a painful, swollen ankle after a cat bite to his leg . On aspiration of the ankle, gram negative pleomorphic rods and monosodium urate crystals were seen and Pasteurella multocida was cultured . He was treated with ampicillin/sulbactam, joint aspiration, and intraarticular steroids, with resolution of infection and return of joint function . The syndromes of Pasteurella arthritis and crystal arthropathy with septic arthritis are reviewed. Am J Vet Res, 1997 Aug, 58(8), 841 - 7 Efficacy of a subcutaneously administered, ultraviolet light-killed Pasteurella multocida A:3-containing bacterin against transthoracic challenge exposure in goats; Purdy CW et al.; OBJECTIVE: To determine the effectiveness of Pasteurella multocida biovar A, serovar 3 (Pm A:3) killed by exposure to UV light and incorporated with a polyacrylate bead carrier as a vaccine . ANIMALS: 18 weanling male Spanish goats . PROCEDURE: Prospective, randomized controlled study with 3 treatment groups: positive-control (PC), negative-control (NC), and principal Pm A:3 bacterin (PA) groups . Six PC goats each received live Pm A:3 and polyacrylate beads twice, 22 days apart, by transthoracic injection into the left lung . Six NC goats each received only PA beads twice, 22 days apart, by transthoracic injection . Six principal goats each received Pm A:3 vaccine SC twice, 22 days apart . Fourteen days after the second vaccination, all goats were challenge exposed with live Pm A:3 by transthoracic injection into the right lung, and 4 days later they were euthanatized and necropsied . RESULTS: Mean volume of consolidated lung tissue at the challenge site was 1.75 cm3 for the PC group, 15.18 cm3 for the NC group, and 3.9 cm3 for the PA vaccine group . The NC group had a significantly (P < or = 0.002) larger mean volume of consolidated lung tissue than did the PC and PA groups after challenge exposure . CONCLUSIONS: The PA bacterin and the PC groups developed protective immunity against live Pm A:3 challenge exposure . An SC administered, UV light-killed, Pm A:3 bacterin induced protective immunity similar to that induced by virulent live Pm A:3 injected into the target organ, the lung. J Clin Microbiol, 1997 Aug, 35(8), 1948 - 51 Evaluation of transport media for Pasteurella multocida isolates from rabbit nasal specimens; Kawamoto E et al.; A suitable medium for the transport of Pasteurella multocida in nasal specimens from rabbits was investigated by using pure cultures of the organism and nasal swabs from infected rabbits . First, the ability of eight transport media to preserve the viabilities of P . multocida strains isolated from rabbits was studied . Cary-Blair medium and Leibovitz medium no . 15 (L-15) were found to be superior to the other six media tested, enabling survival of the organism for more than 14 days at room temperature . Second, the survival of P . multocida in nasal specimens was evaluated on both Cary-Blair medium and L-15 . The recovery rate of the organism from these two media was more than 80 to 90% during 4 days of storage and decreased gradually with increasing preservation time . There were no significant differences (P > 0.05) in recovery rates of the organism between Cary-Blair medium and L-15 . On the basis of these results, we recommend the use of Cary-Blair medium for the transport of P . multocida in rabbit nasal specimens because of the ease of transport of nasal swabs by mail. Vet Rec, 1997 Jul 12, 141(2), 37 - 40 Comparative in vitro activity of doxycycline and oxytetracycline against porcine respiratory pathogens; Bousquet E et al.; Minimum inhibitory concentrations of doxycycline and oxytetracycline were determined against 55 Pasteurella multocida strains, 59 Actinobacillus pleuropneumoniae strains and 26 Mycoplasma hyopneumoniae strains isolated from the respiratory tract of pigs . An additional set of 76 P multocida strains isolated from pneumonic pigs was tested for their minimum inhibitory concentrations of doxycycline . The P multocida and A pleuropneumoniae strains were isolated in France and the minimum inhibitory concentrations were determined by an agar dilution method . The M hyopneumoniae strains were isolated in the United Kingdom and minimum inhibitory concentrations were determined by a serial broth dilution method . All the strains tested were susceptible to doxycycline whereas 15 per cent of the P multocida strains and 22 per cent of the A pleuropneumoniae strains were resistant to oxytetracycline . Doxycycline concentrations inhibiting 90 per cent of strains were 1 microgram/ml for P multocida and 2 micrograms/ml for A pleuropneumoniae . The ratio of the minimum inhibitory concentrations of doxycycline and oxytetracycline ranged between 1/1 and 1/4 for the oxytetracycline-susceptible strains and between 1/16 and 1/64 for the oxytetracycline-resistant strains . All the M hyopneumoniae strains were susceptible to doxycycline and oxytetracycline, the concentrations inhibiting 90 per cent of strains being 1 microgram/ml and 2 micrograms/ml, respectively . These data confirm that doxycycline has a higher in vitro activity against pig respiratory pathogens than oxytetracycline. J Comp Pathol, 1997 Jul, 117(1), 73 - 82 Effects of Trypanosoma evansi on the output of cells from a lymph node draining the site of Pasteurella haemolytica vaccine administration; Onah DN et al.; The prefemoral efferent lymphatics of sheep infected with Trypanosoma evansi and inoculated with P . haemolytica vaccine and of those given only the vaccine, were surgically cannulated to study the effects of the infection on the total cellular output and output of blast cells from the node in response to the vaccine . T . evansi delayed and depressed the increases in total cell and lymphoblast outputs . In uninfected sheep, the total cellular output increased and peaked at more than twice the prevaccination values on days 4 and 5 after primary vaccination, but the increases were smaller and peaked on days 6 and 8 after primary vaccination in the infected sheep . The output of lymphoblasts mirrored the total cell output, though it was suppressed to a greater degree by T . evansi . The output of blasts peaked at more than 8 and 14 times the prevaccination values in the uninfected animals after primary and secondary (booster) vaccinations, respectively; but in infected animals, it peaked at twice the prevaccination values after the primary vaccination and showed no increase after booster vaccination until 11 days later . It is concluded that the inhibition of total and blast cell outputs by T . evansi may limit the early systemic dissemination of antigen-specific cells, thus playing a role in the induction of immunosuppression by the parasite. Vet Immunol Immunopathol, 1997 Jul, 57(3-4), 279 - 87 Heterogeneity of porcine alveolar macrophages in experimental pneumonia; Berndt A et al.; The aim of the study was the morphological and the phenotypic characterization of the porcine non-lymphocytic bronchoalveolar lavage (BAL) cell population of unaffected- and intrabronchial with Pasteurella multocida- (P.m.) infected swine using flow cytometry . Three non-lymphocytic cell populations of the porcine bronchoalveolar lavage could be differentiated: (1) large, high autofluorescent cells, (LHC); (2) small, high autofluorescent cells, (SHC); (3) small, low autofluorescent cells, (SLC) . In comparison with the control animals, the percentage of the LHC and SHC within the whole non-lymphocytic cell population was decreased, whereas the SLC was significantly enhanced after infection . In order to investigate the phenotype of these cell populations, monoclonal antibodies against porcine antigens (SWC1, SWC3a, MHC class II, 2G6 (against macrophages)) were used . The results showed that the cells of the SLC seem to belong to the granulocytes, whereas the LHC and the SHC are lung macrophages . After the infection of the animals the percentage of the SWC1 positive cells of LHC and SHC were significantly increased, indicating an entrance of more immature macrophages . The percentage of the MHC class II antibody binding cells of all three non-lymphocytic populations was-decreased after infection, indicating a restricted MHC class II dependent antigen recognition in P.m . pneumonia. J Wildl Dis, 1997 Jul, 33(3), 558 - 66 Effect of simulated stress on susceptibility of bighorn sheep neutrophils to Pasteurella haemolytica leukotoxin; Kraabel BJ et al.; We examined the effects of simulated stress on susceptibility of Rocky Mountain bighorn sheep (Ovis canadensis canadensis) neutrophils to Pasteurella haemolytica leukotoxin in a blocked, crossover experiment . Ten captive-raised bighorn sheep were sampled 10 hr after separate administrations of long-acting adrenocorticotrophic hormone (ACTH) gel and normal saline (control) . We then compared in vitro leukotoxin-dependent neutrophil death rates after exposure to culture supernatants from four unique P . haemolytica isolates (one from domestic and three from bighorn sheep) . Simulated stress effects were evidenced by elevated (P = 0.002) mean plasma cortisol concentrations, more neutrophils (P = 0.037), and fewer lymphocytes and eosinophils (P < or = 0.043) in ACTH-treated bighorn sheep . Maximum leukotoxin-dependent neutrophil death rates were > or = 61% for three of four P . haemolytica isolates tested . For all three cytotoxic isolates, neutrophil death rates at 150 micrograms/50 microliters supernatant were about 1.13 times higher (P = 0.0001) after bighorns received ACTH; for two of these, overall neutrophil death rates were higher (P < or = 0.001) in ACTH-treated bighorn sheep . Although variable leukotoxin production among P . haemolytica strains appeared principally responsible for differences in leukotoxin-dependent neutrophil death rates, susceptibility of bighorn sheep neutrophils to leukotoxin was increased by prior exposure to elevated plasma cortisol concentrations . It follows that if similar processes occur in neutrophils and alveolar macrophages in vivo, they could contribute to greater susceptibility of stressed bighorn sheep to pneumonic pasteurellosis. J Wildl Dis, 1997 Jul, 33(3), 544 - 57 Pasteurella spp . in sympatric bighorn and domestic sheep; Ward AC et al.; Domestic sheep were sighted at different times from 1991 to 1993 on four Nevada (USA) ranges occupied by bighorn sheep . Nasal and pharyngeal swab samples were collected from both sheep species and cultured to determine if any strains of Pasteurella spp . were shared on range conditions after contact of the two species . Pasteurella spp . were isolated from all 38 bighorn sheep and 16 of 17 domestic sheep included in this study . The isolates were characterized on the bases of species, biotype, serotype, biogroup, and restriction enzyme analyses (REA) as well as ribotyping of bacterial DNA . A P . haemolytica biotype 3, biogroup 11 isolate from a domestic sheep had biochemical, REA, and ribotype profiles which were identical to those of isolates from three bighorn sheep on the same range . None of the other isolates were found to be common to the two sheep species . Disease was not detected in any of the bighorn populations . However, bighorn sheep populations were extirpated on two ranges while increasing on the other two, including the range on which P . haemolytica biotype 3, biogroup 11 strain was isolated . Declining sheep numbers were not correlated with the presence of any one strain of Pasteurella spp from the sheep. Toxicon, 1997 Jul, 35(7), 999 - 1010 Sir Charles James Martin MB FRS: Australian serpents and Indian plague, one-hundred years ago; Hawgood BJ; In 1891 as Demonstrator in Physiology at the University of Sydney, Charles Martin began the first systematic study of the chemical and physiological properties of the venoms of the Australian elapid species, Pseudechis porphyriacus and Notechis scutatus . Two major constituents were detected: a large coagulable protein which was associated with intravascular clotting, and a small proteinaceous molecule, an albumose, associated with neurotoxicity . Martin designed and constructed a high-pressure gelatin membrane ultrafilter for fractionation of venom . His studies indicated that certain physiological actions and clinical symptoms were related to the faster rate of diffusion within the tissue space of a neurotoxic constituent relative to a clotting constituent . Extending this work to toxin-antitoxin relationships, Martin provided evidence that antitoxin was a large molecule with slow diffusibility in tissue and advised the administration of curative serum (including diphtheria antitoxin) by intravenous injection . In 1903, Martin returned to London as Director of the Lister Institute of Preventive Medicine . He was soon involved in the planning of scientific work to be undertaken by the Commission for Investigation of Plague in India as the disease continued to ravage the subcontinent . Detailed epidemiological studies of possible factors involved in the spread of Pasteurella pestis showed, unequivocally, that infected rat fleas were the vector of transmission from rats to humans. Can J Vet Res, 1997 Jul, 61(3), 187 - 92 Treatment of experimentally induced pneumonic pasteurellosis of young calves with tilmicosin; Morck DW et al.; Twenty four (24) healthy male Holstein calves (< 70 kg) were each experimentally infected by intrabronchial inoculation of 4.0 x 10(9) viable cells of Pasteurella haemolytica-AI (B122) at Time = 0 h . At 1 h following inoculation animals received either: 1) Sham treatment with sterile 0.85% saline SC (n = 12); or 2) a single injection of 10 mg tilmicosin per kg body weight (n = 12) . Calves that were non-infected and tilmicosin-treated were also included for determining tilmicosin concentrations in serum and lung tissue at 1, 2, 4, 6, 8, 24, 48, and 72 h (n = 3-per time) . In the infected calves, response to therapy was monitored clinically . Serum samples were collected for determination of tilmicosin concentrations using HPLC . Any animal becoming seriously ill was humanely killed . Complete necropsy examinations were performed on all animals and included gross pathologic changes, bacteriologic analysis, histopathology, and determination of pulmonary concentrations of tilmicosin . Tilmicosin treated animals responded significantly better to therapy than saline-treated control calves . Clinical assessment of calves during the study indicated that tilmicosin-treated calves had significantly improved by T = 8 h compared to satine-treated animals (P < 0.05) . At necropsy tilmicosin-treated calves had significantly less severe gross and histological lesions (P < 0.05) of the pulmonary tissue . Of the 12 saline-treated calves, 92% (11/12) had Pasteurella haemolytica-A1 in lung tissue, while of the tilmicosin-treated calves 0% (0/12) cultured positive for P . haemolytica . Mean (+/- standard error) serum tilmicosin concentrations in infected calves peaked at 1 h post-injection (1.10 +/- 0.06 micrograms/mL) and rapidly decreased to 0.20 +/- 0.03 microgram/mL, well below the MIC of 0.50 microgram/mL for P . haemolytica-A1 (B122), by 12 h . These serum concentrations were very similar to serum concentrations of tilmicosin in non-infected tilmicosin-treated calves . Lung tissue concentrations of the antibiotic were comparatively high, even at 72 h post-infection (6.50 +/- 0.75 ppm) . Lung tissue concentrations at 72 h were significantly higher in experimentally infected calves than in non-infected tilmicosin-treated animals (P < 0.05) . These data demonstrate that tilmicosin was effective in treating experimentally-induced pneumonic pasteurellosis as determined by alleviation of clinical signs, pathological findings at post mortem, and presence of viable bacteria from the lung . Concentrations substantially above MIC for P . haemolytica were present in lung tissue even at 72 h following a single subcutaneous injection of 10 mg tilmicosin per kg body weight. J Med Microbiol, 1997 Jul, 46(7), 603 - 10 Identification by monoclonal antibodies of serotype D strains of Pasteurella multocida representing various geographic origins and host species; Vasfi Marandi M et al.; Two outer-membrane proteins (OMPs) of Pasteurella multocida serotype D, designated H and W, possess potentially important serotype D-specific antigens . Antigenicity as well as toxigenicity of 55 strains of P . multocida representing various serotypes, geographic origins and host species were studied by SDS-PAGE, enzyme-linked immunosorbent assay (ELISA), immunoblot and polymerase chain reaction (PCR) assays . Based on the electrophoretic mobility of protein H, different OMP patterns were observed within different capsular serotypes . Three monoclonal antibodies (MAbs) designated MT1, MT2 and MT3 were produced against H and W proteins of P . multocida in BALB/c mice . MAbs MT2 and MT3 reacted with two distinct epitopes on W protein of serotype D in competitive ELISA . MAb MT1 reacted with all serotype D-I strains but not with D-II strains, whereas MAb MT2 reacted with both serotype D-I and D-II strains in dot-ELISA and immunoblot assay . MAb MT3 reacted with all P . multocida strains belonging to different capsular serotypes in dot-ELISA . None of the MAbs reacted with other gram-negative bacteria tested, indicating that protein H has a serotype D-I specific epitope and protein W has both serotype and species-specific epitopes . PCR assay was used to identify toxigenic strains of P . multocida; 92% of P . multocida strains possess both toxA gene and MAb MT2 reacting epitope, suggesting a strong association between MAb MT2 reacting epitopes and toxA gene . Rapid dot-ELISA with MAb was found to be specific, sensitive and easy to perform and thus suitable for routine serotyping of P . multocida serotype D strains which might be potentially pathogenic. Arch Otolaryngol Head Neck Surg, 1997 Jul, 123(7), 759 - 61 Pasteurella multocida epiglottitis; Wine N et al.; Pasteurella multocida, a small gram-negative coccobacillus, colonizes the nasopharynx and gastrointestinal tract of many animals, including cats and dogs . Most human infections with P multocida are due to animal bites, but the respiratory tract is the second most common site of infection . We describe the third case report (to out knowledge) of acute P multocida epiglottitis . The mode of transmission in this case was inhalation of infectious nasopharyngeal secretions from cats . The patient responded well to treatment with penicillin, the drug of choice for P multocida infections . Therefore, infection with P multocida, though rare, should be considered in the differential diagnosis in any case involving acute epiglottitis and exposure to cats. Lab Anim, 1997 Jul, 31(3), 193 - 200 Microbiological monitoring of laboratory pigs; Hansen AK et al.; Purpose-bred minipigs, are often used as the non-rodent species in toxicology . Infections may interfere with animal experiments, and there are no scientific reasons why the non-rodent species should be of a lower microbiological quality than the rodent species . Therefore, a system for health monitoring of pigs was developed in order to raise the quality of laboratory pigs to the level of laboratory rodents . This system, which includes screening for several viruses, bacteria and ecto- and endoparasites, was used for monitoring minipigs from a barrier unit with the same standards applied to rodents units . In these pigs only rotaviruses are found, which was shown by both serological antibody detection and by detection of rotaviral antigen in faeces . In minipigs from another unit with far less hygienic protection rotaviruses were also found along with certain influenza- and coronaviruses, as well as Pasteurella spp . It is concluded, that it is possible to raise pigs of a microbiological quality comparable to the quality of rats and mice, and that advanced microbiological monitoring in pigs will reveal useful information. Am J Vet Res, 1997 Jul, 58(7), 755 - 9 Intrastrain variation of lipopolysaccharide of Pasteurella multocida in turkeys; Coy SL et al.; OBJECTIVE: To document intrastrain variation of lipopolysaccharide (LPS) in Pasteurella multocida and correlate these changes with changes in determinants associated with virulence . ANIMALS: 25 broad-breasted white turkeys . PROCEDURE: Phenotypic bacterial variants were identified by lectin affinity and were assayed for adherence to epithelial cells and complement resistance in vitro . The LPS purified from these variants was subjected to polyacrylamide gel electrophoresis and lectin affinity analysis . Turkeys were challenge exposed, then observed for 1 week . At first sign of disease, or at the end of the study, turkeys were euthanatized, necropsied, and inspected for gross lesions . RESULTS: The LPS variant designated as Ricinus communis agglutinin-positive had greater adherence to epithelial cells, complement resistance, and virulence in turkeys than did the variant designated as R communis agglutinin-negative . CONCLUSIONS: Intrastrain variation of LPS exists in P multocida, and changes in LPS are correlated with changes in virulence. Am J Vet Res, 1997 Jul, 58(7), 749 - 54 Growth of Pasteurella haemolytica and production of its leukotoxin in semi-defined media; Highlander SK; OBJECTIVE: To develop semi-defined media that support growth of the bovine pathogen, Pasteurella haemolytica, and use them to examine production of leukotoxin and an arginine-binding protein by this organism . SAMPLE POPULATION: 10 P haemolytica A1 strains and 1 P multocida strain . PROCEDURE: Bacterial strains were cultivated at 37 C in media containing various amino acids, carbon sources, vitamins, and cofactors, and absorbance (OD600) was measured . Leukotoxin and arginine-binding protein production were assessed by immunoblot analysis . RESULTS: Optimal growth required supplementation with 0.1% fetal bovine serum, gelatin, or purified bovine serum albumin . Calcium pantothenate and thiamine were essential for growth, and a variety of carbon sources could be utilized . In the complete medium, 15 amino acids were included; however, in the minimal medium, no amino acids were required . All strains (except P multocida) grew in the complete medium and 7 grew well in the minimal medium . Leukotoxin was not produced when amino acids were limiting, but could be enhanced by addition of 0.2% NH4SO4 . Production of the arginine-binding protein was not affected by nitrogen availability or by presence of L-arginine . CONCLUSIONS: Two media that support good growth of P haemolytica strains were developed . The minimal medium is simple to prepare and manipulate and its use revealed a potential role of nitrogen availability in the regulation of leukotoxin expression . CLINICAL RELEVANCE: Creation of these media will permit continued studies of the response of P haemolytica to environmental conditions that may mimic those encountered in the bovine respiratory tract during shipping. Mol Microbiol, 1997 Jun, 24(5), 1061 - 70 Molecular localization of the Escherichia coli cytotoxic necrotizing factor CNF1 cell-binding and catalytic domains; Lemichez E et al.; Cytotoxic necrotizing factor type 1 (CNF1) induces, in epithelial cells, the development of stress fibres via the GTPase Rho pathway . We showed that CNF1 is able to modify Rho both in vitro and in vivo . Recombinant N-terminal 33kDa (CNF1Nter) and C-terminal 14.8-31.5 kDa (CNF1Cter) regions of the CNF1 protein allowed us to demonstrate that the N-terminal region contains the cell-binding domain of the toxin and that the C-terminal region is responsible for its catalytic activity . CNF1Nter lowered the activity of CNF1 when provided to cells before the toxin whereas CNF1Cter had no effect on CNF1 cell toxicity . CNF1Cter was sufficient to induce a typical CNF1 phenotype when microinjected into African green monkey kidney cells (Vero cells), and was able to modify Rho as previously reported for CNF1 . The C-terminal domain lost its catalytic activity when deleted of various subdomains, suggesting a scattered distribution of catalytic-site amino acids . Elucidation of the CNF1 functional organization and analysis of amino acid homologies between CNFs (CNF1, CNF2), Pasteurella multocida toxin (PMT) and dermonecrotic toxin of Bordetella pertussis (DNT) allowed us to postulate that CNFs and DNT act on Rho via the same enzymatic activity located in their C-terminus, and that CNFs and PMT probably bind to analogous cell receptors. Pediatr Nephrol, 1997 Jun, 11(3), 353 - 4 Pasteurella multocida peritonitis in patients undergoing peritoneal dialysis; Loghman-Adham M; A 12-year-old girl on peritoneal dialysis developed sub-clinical peritonitis due to Pasteurella multocida, following puncture of her dialysis tubing by a domestic cat . Only four other similar cases of P . multocida peritonitis have been reported in adults . This unusual form of peritonitis could be easily prevented by not allowing domestic animals to come into contact with dialysis tubings in patients undergoing peritoneal dialysis. FEMS Microbiol Lett, 1997 May 15, 150(2), 197 - 202 Divergent activity and function of superoxide dismutases in Pasteurella haemolytica serotypes A1 and A2 and Pasteurella trehalosi serotype T10; Rowe HA et al.; Representative strains of Pasteurella haemolytica serotypes A1 and A2 and Pasteurella trehalosi serotype T10 were examined for the presence of superoxide dismutase . Visualisation of superoxide dismutase enzyme activity on polyacrylamide gels, and specific inhibition with potassium cyanide verified a copper/zinc (Cu/Zn) superoxide dismutase only in serotype A2 whereas serotypes A1 and T10 showed other superoxide dismutase activity . Using a simple freeze-thaw method the cellular location of superoxide dismutase enzyme activity was determined in all three serotypes . In serotypes A1 and A2 but not T10 superoxide dismutases were located in the periplasm . The viability of serotypes A2 and T10 cells in the presence of exogenous superoxide was unchanged over a 30 min period, whereas serotype A1 cells declined in viability between 15 and 30 min . Purified immunoglobulin from sheep convalescent serum did not reduce superoxide dismutase activity in the serotypes in an in vitro assay . The presence of this enzyme within the pasteurellae suggests a supportive role in the virulence of this major pathogen of ruminants. J Infect, 1997 May, 34(3), 263 - 4 Pasteurella multocida infection of a total hip arthroplasty following cat scratch; Takwale VJ et al.; Pasteurella multocida is a well recognized cause of sepsis following animal contact particularly bites and scratches . Spread to prosthetic joints may occur particularly in immunocompromised patients . Immunocompromised patients with prosthetic joints should be warned that animals are potential sources of serious infection and urgent medical advice should be sought if bitten or scratched. Zentralbl Veterinarmed A, 1997 May, 44(3), 179 - 87 Efficacy of the combination sodium ceftiofur-flumethasone in the treatment of experimental Pasteurella haemolytica bronchopneumonia in calves; Sustronck B et al.; Severe acute bronchopneumonia was induced in 18 conventional Friesian-Holstein calves by inoculating them intratracheally with Pasteurella haemolytica type A1 . Six of the calves received no treatment and served as controls . Six of the calves were treated with sodium ceftiofur and six were treated with sodium ceftiofur and flumethasone . The mortality rate in the group of calves treated with sodium ceftiofur and flumethasone was significantly lower and their clinical and haematological parameters returned to normal significantly faster than in the control calves and the calves treated with sodium ceftiofur alone. J Biochem (Tokyo), 1997 May, 121(5), 902 - 13 Two genes encoding serine protease homologues in Serratia marcescens and characterization of their products in Escherichia coli; Ohnishi Y et al.; A serine protease (SSP) of Serratia marcescens is one of the extracellular enzymes secreted from this Gram-negative bacterium . SSP is produced as a large precursor and converted to a mature protein by cleavages removing an NH2-terminal signal sequence and a COOH-terminal pro-region . This COOH-terminal pro-region is integrated into the outer membrane and has a functional role for the export of the mature protein across the outer membrane . Southern hybridization analysis with a DNA fragment encoding the COOH-terminal pro-region as the probe showed a wide distribution of nucleotide sequences encoding SSP exporter-like proteins among Serratia species . Moreover, S . marcescens IFO 3046, from which the ssp gene had been cloned, was found to contain two ssp homologues (ssp-h1 and ssp-h2) . They were cloned and their nucleotide sequences were determined . The two ssp homologues were found to exist in tandem on the genome and their amino acid sequences showed 81% identity to each other . Both of them showed 55% identity in amino acid sequence to preproSSP . In addition, both showed end-to-end similarity to the 100 kDa serotype-specific antigen (Ssa1) of Pasteurella haemolytica . Escherichia coli JM105 containing ssp-h1 gene produced a 53 kDa protein corresponding to the NH2-terminal portion and a 49 kDa protein corresponding to the COOH-terminal portion, both of which were rigidly integrated in the outer membrane . Consistent with the significant similarity of the COOH-terminal portions of the homologues to that of SSP, they showed the ability to translocate the mature SSP part across the outer membrane into the medium . Furthermore, the NH2-terminal portion of the homologue was not translocated into the outer membrane without its COOH-terminal part . All of these data show that the SSP homologues are outer membrane proteins that are translocated into the outer membrane with the aid of the translocator function of their COOH-terminal part. J Comp Pathol, 1997 May, 116(4), 415 - 8 Pulmonary venous thrombosis in caprine Pasteurella pneumonia; Scholes SF et al.; A 3-year-old Angora goat that developed acute fibrinous pleuropneumonia associated with Pasteurella haemolytica infection had thrombotic occlusion of a large pulmonary vein . Thrombosis of pulmonary capillaries occurs in pneumonic pasteurellosis, but large vessels are not commonly affected . This unusual lesion may reflect the procoagulant effect of pasteurella endotoxin on vascular endothelium . An incidental observation was the presence of myocardial-type muscle fibres in the tunica media of the pulmonary vein. Antimicrob Agents Chemother, 1997 May, 41(5), 1193 - 5 Comparative in vitro activities of DU-6859a, levofloxacin, ofloxacin, sparfloxacin, and ciprofloxacin against 387 aerobic and anaerobic bite wound isolates; Goldstein EJ et al.; The activities of DU-6859a, levofloxacin, ofloxacin, sparfloxacin, and ciprofloxacin against bite wound isolates were determined by the agar dilution method . DU-6859a was the most active compound (MICs, < or = 0.125 microg/ml) against all Pasteurella species, Staphylococcus aureus, and streptococci; anaerobes were susceptible to < or = 0.5 microg/ml, except fusobacteria, which were susceptible to < or = 2 microg/ml . Against aerobes, levofloxacin was more active than ofloxacin (MIC at which 90% of isolates are inhibited {MIC90}, < or = 1.0 microg/ml for both) and sparfloxacin and ciprofloxacin were also active (MIC90s, < or = 0.25 and < 1 microg/ml, respectively). Clin Infect Dis, 1997 May, 24(5), 1004 - 6 Use of random amplification of polymorphic DNA in a case of Pasteurella multocida meningitis that occurred following a cat scratch on the head; Schuur PM et al.; We cultured Pasteurella multocida from the cerebrospinal fluid (CSF) of a 4-month-old infant who presented with meningitis . The patient had been scratched on the head by a cat . Culture of the cat's claws also yielded P . multocida . The isolates had identical biochemical patterns . Analysis of both strains by random amplification of polymorphic DNA and comparison of these strains with P . multocida strains isolated from other cats showed that the two strains were identical and completely different from the unrelated isolates . Our patient's meningitis most likely resulted from direct inoculation of P . multocida into the CSF. J Med Chem, 1997 Apr 25, 40(9), 1340 - 6 Quantitative structure-activity relationships among macrolide antibacterial agents: in vitro and in vivo potency against Pasteurella multocida; McFarland JW et al.; Quantitative structure-activity relationships have been found among macrolide antibacterial agents in their potencies against the bacterial pathogen Pasteurella multocida both in vitro and in mouse infections . To obtain these relationships we measured, among other things, the pK(a)'s and log P's of 15 known macrolides of diverse structures . Among these compounds, in vitro potency {log(1/MIC)} is a function of log P, log D, and CMR (R = 0.86) . In vivo potency is a function of the higher pK(a), the HPLC chromatographic capacity factor log k', log(1/MIC) and pNF (R = 0.93) . pNF is defined as the negative logarithm of the fraction of neutral drug molecules present in aqueous solution at pH 7.4 . The same physical properties were determined for 14 macrolides not used in developing the original QSAR models . Using the in vivo model, we calculated the mouse protection potency ranges for these new compounds . Ten estimates agreed with those observed, three were lower by a half-order of magnitude, and one was calculated to be active in the range of 15-50 mg/kg, but in fact was not active at 50 mg/kg, the highest level tested . When these new compounds were combined with the original 15, and the QSAR's updated, the new equations for the in vitro and in vivo potencies were essentially the same as those originally found . Hence, the physical properties indicated above are major determinants of macrolide antibacterial potencies. Vet Microbiol, 1997 Apr, 55(1-4), 241 - 6 Effect of porcine reproductive and respiratory syndrome virus on subsequent Pasteurella multocida challenge in pigs; Carvalho LF et al.; This trial was conducted to evaluate the effect of Porcine reproductive and respiratory syndrome virus (PRRSv) on a subsequent challenge with Pasteurella multocida in pigs . Sixteen, 3-4 week-old piglets, from a PRRSv and Aujeszky disease virus (ADV) free herd were used . Animals were equally and randomly allocated in four groups which were treated according the following schedule: Group I: negative controls; Group II: inoculation with only PRRSV; Group III: inoculation with PRRSV and P . multocida; Group IV: inoculation with ADV and P . multocida (positive controls) . PRRSV and ADV were inoculated intranasally, at the doses of 10(4.6) and 10(4.5) TCID50/ml, respectively . Five days later, pigs from groups III and IV were inoculated intranasally, with two ml of a 10(9) CFU/ml suspension of equal parts of P . multocida, strains A52 and A24 . No lesions were observed in piglets of group I . Microscopically, interstitial pneumonia was identified in all piglets of groups II and III and 3/4 piglets from group IV . Bronchopneumonia was detected in 3/4 of the piglets from group III and in all animals of group IV which, additionally, showed meningo-encephalitis and purulent rhinitis . Macroscopically, only piglets of groups III and IV had lung consolidation . However, much lower pneumonic scores (2.3%) were observed in group III, where 3 of 4 piglets were affected . On the other hand, all piglets of group IV showed some degree of pulmonary consolidation, with a mean score of 13.7% . Based on these results, it appears that the role of PRRSV as a initiator of secondary diseases is still undefined, but is probably mild . There was no clear interaction between PRRSv and Pasteurella multocida under the conditions and strains tested here. Avian Dis, 1997 Apr-Jun, 41(2), 317 - 25 Comparison of live avirulent PM-1 and CU fowl cholera vaccines in turkeys; Hopkins BA et al.; The live avirulent PM-1 Pasteurella multocida vaccine, grown in brain-heart infusion broth, was evaluated and compared in two experiments with the Clemson University (CU) vaccine, which had been shown to be effective in preventing fowl cholera in turkeys . Experiment 1 was performed during warm environmental temperatures and Expt . 2 during cooler environmental temperatures . The PM-1 vaccine was comparable with the CU vaccine in protecting turkeys against challenge with virulent P . multocida but was considered no less virulent than the CU because turkeys died after vaccination with both the PM-1 and the CU vaccines . A significantly (P < 0.05) higher percentage of unvaccinated turkeys challenged during the cooler environmental temperatures died than did unvaccinated turkeys challenged during the warmer temperatures . A microtiter agglutination test demonstrated a significant (P < 0.01) correlation between the level of serum anti-P . multocida antibody found 1 wk after vaccination and survival after challenge with virulent P . multocida in Expt . 1 and a significant (P < 0.05) correlation between these parameters in Expt . 2 . However, there was a significant (P < 0.01) negative correlation between serum anti-P . multocida antibody titer 1 wk after vaccination and body weight gained 4 wk after vaccination, but before challenge, in Expt . 1, suggesting that vaccination with the live vaccines may have had a negative effect on body weight gain . At 4 wk after challenge or 8 wk after vaccination in Expt . 2, there was also a highly significant (P < 0.001) negative correlation between these parameters in the surviving turkeys. Berl Munch Tierarztl Wochenschr, 1997 Apr, 110(4), 139 - 42 {Detection of dermonecrotic toxin genes in Pasteurella multocida strains using the polymerase chain reaction (PCR)}; Hotzel H et al.; A PCR method was developed which allows to distinguish between Pasteurella multocida strains carrying or lacking the dermonecrotic toxin gene . Specific primers were used to amplify a 1501-bp DNA fragment from the genomic dermonecrotic toxin gene region . Isolated DNA, broth cultures and swabs were used as samples . Detection of the toxin gene directly from swab samples accelerates considerably the diagnosis since cultivation steps can be omitted . The results of PCR corresponded to findings obtained by ELISA. J Trace Elem Med Biol, 1997 Apr, 11(1), 28 - 31 Udder orf infection and its role in ovine clinical mastitis caused by Pasteurella haemolytica; Burriel AR; During an experimental study of ovine subclinical mastitis caused by coagulase-negative staphylococci, an outbreak of contagious ecthyma occurred among ewes unvaccinated against parapox virus . The same group of ewes developed a high rate (43.7%) of clinical mastitis caused by Pasteurella haemolytica . The rate of clinical mastitis among ewes vaccinated against parapox virus was very low (3.7%) suggesting that the presence of orf in the unvaccinated ewes was contributing to the high rate of clinical mastitis . An examination of the iron, sodium, potassium and albumin concentration of milk collected from 16 unvaccinated and nine randomly selected vaccinated ewes before experimental infection with coagulase-negative staphylococci or their uninfected control mammary glands indicated significant differences in the iron (p < 0.0001) and sodium (p = 0.01) concentration . Increased iron concentration in the milk may have assisted in the development of udder infection caused by P . haemolytica as iron is easily utilised by this bacterium. Zentralbl Veterinarmed B, 1997 Apr, 44(2), 99 - 104 Bacteria associated with enzootic pneumonia in goats; Oros J et al.; A histological and microbiological study of lung samples from 83 slaughtered goats (33 kids and 50 adults) drawn from a flock with a history of pleuropneumonia caused by mycoplasmas of the M . mycoides group was carried out . A total of 82% (27/33) of kids and 36% (18/50) of adult goats presented pulmonary lesions characteristic of enzootic pneumonia: lesions took the form of bronchointerstitial pneumonia with peribronchial and peribronchiolar proliferation of lymphocytes . Microbiological analysis confirmed a range of mycoplasma species, including Mycoplasma mycoides ssp . mycoides Large Colony (MmmlC) (3.70%; 1/27), Mycoplasma mycoides ssp . capri (Mmc) (7.40%; 2/27), Mycoplasma putrefaciens (22.2%; 6/27), Mycoplasma arginini (3.70%; 1/27) and Mycoplasma sp . (7.40%; 2/ 27), as well as Pasteurella multocida (14.8%; 4/27), associated with enzootic pneumonia lesions in younger animals, whereas Mycoplasma sp . was associated with enzootic pneumonia in adult goats (22.0%; 4/18) . Cilia-associated respiratory (CAR) bacillus found by histochemical examination was associated with enzootic pneumonia in kids (25.9%; 7/27) and goats (44.4%; 8/18), being the first description of this bacterium in adult goats. Zentralbl Bakteriol, 1997 Apr, 285(4), 459 - 79 Relationships among strains classified with the ruminant Pasteurella haemolytica-complex using quantitative evaluation of phenotypic data; Angen O et al.; The phenotypic relationships among 246 trehalose-negative strains classified under the {Pasteurella} haemolytica-complex in ruminants were investigated by clustering and multidimensional ordinations based upon 79 phenotypic characters . A quantitative evaluation of phenotypic data using a 5-level scoring system is presented permitting a comprehensive utilization of the recorded phenotypic variation among the strains in the analyses . Clustering and ordination analyses display complementary aspects of data which has been clearly demonstrated in this investigation . The main clusters revealed by the numerical techniques could be related to distinctive phenotypic differences and showed an extensive correlation with the recognized biogroups . This classification was based only upon 4 characters (fermentation of L-arabinose, D-sorbitol, glucosides and ornithine decarboxylase) . In contrast, there was no obvious interpretation of the clusters formed by using binary scores . Phenotypic subgroups within the recognized biogroups have been described as well as a new, related group of bacteria, tentatively named Bisgaard taxon 36 . Quantitative interpretation of phenotypic data seems to represent a promising method for finding relations among affiliated strains of bacteria and to assist in forming hypotheses for subsequent genotypic investigations. J Wildl Dis, 1997 Apr, 33(2), 332 - 5 Pasteurella multocida serotype 1 isolated from a lesser snow goose: evidence of a carrier state; Samuel MD et al.; Pharyngeal swabs were collected from 298 lesser snow geese (Chen caerulescens caerulescens) at Banks Island (Northwest Territories . Canada) in the summer of 1994 . Pasteurella multocida serotype 1 was isolated from an adult male bird and P . multocida serotype 3 was isolated from an adult female goose . Pathogenicity of the serotype 1 isolate was confirmed by inoculation in Pekin ducks (Anas platyrhynchos) . The serotype 3 isolate was non-pathogenic in Pekin ducks . This is the first documented isolation of pathogenic P . multocida serotype 1 from apparently healthy wild snow geese. J Med Microbiol, 1997 Apr, 46(4), 276 - 84 Characterisation of the leukotoxin produced by different strains of Pasteurella haemolytica; Saadati M et al.; Pasteurella haemolytica isolates from cattle and sheep, including representatives of different serotypes and untypable strains, were examined for leukotoxin (Lkt) production at the end of the log phase of growth in brain heart infusion broth . There were marked differences in leukotoxic activity in culture supernate samples, as measured by chemiluminescence-inhibition assays with bovine and ovine neutrophils, even between strains of the same serotype . There was also some variation in the amount and mol . wt of the Lkt protein produced by different strains, as judged by SDS-PAGE, immunoblotting and ELISA . Some strains produced normal amounts of Lkt protein which had only low leukotoxic activity . Most strains produced Lkt of 105 kDa whereas four strains produced a higher mol . wt form of c . 108 kDa, including two of the five serotype A2 strains examined . Thus, the P . haemolytica isolates showed considerable heterogeneity in terms of leukotoxin production, mol . wt and activity, even within a given serotype. J Anim Sci, 1997 Apr, 75(4), 1112 - 8 Effects of copper deficiency and copper deficiency coupled with high dietary iron or molybdenum on phagocytic cell function and response of calves to a respiratory disease challenge; Gengelbach GP et al.; A study was conducted to determine the effects of supplementing a diet marginally deficient in copper (Cu) with iron (Fe), molybdenum (Mo), or Cu on phagocytic cell function and disease resistance of calves . Thirty-one calves were born to heifers fed a corn silage-based diet containing 4.5 mg of Cu/kg . Treatments consisted of 1) control (CON; no supplemental Cu, Fe, or Mo), 2) 600 mg of Fe added/kg (FE), 3) 5 mg of Mo added/kg (MO), or 4) 10 mg of Cu added/kg of DM (CU) . Activity of superoxide dismutase was lower (P < .06) in neutrophils from MO vs CON or CU calves at 170 d of age . bactericidal activity of neutrophils from MO calves tended (P = .15) to be lower compared with those from CU calves at 70 d of age . Calves were inoculated intranasally with live infectious bovine rhinotracheitis virus (IBRV) 2 d after weaning, followed by intratracheal administration of Pasteurella hemolytica 5 d later . Iron- and Cu-supplemented calves exhibited higher (P < .01) body temperatures and lower (P < .06) feed intakes following IBRV inoculation compared with CON and MO calves . Copper-supplemented calves had higher levels of plasma tumor necrosis factor (TNF) than MO calves at weaning (P < .05) and tended to have higher plasma TNF (P = .11) than FE and MO calves 5 d after IBRV inoculation . These data indicate that dietary levels of Mo and Cu can affect body temperature and feed intake responses to disease by affecting TNF and perhaps other cytokines. Int J Syst Bacteriol, 1997 Apr, 47(2), 575 - 6 Characterization of leptospiral serovars by randomly amplified polymorphic DNA fingerprinting; Ramadass P et al.; Randomly amplified polymorphic DNA (RAPD) fingerprinting of 14 laboratory strains of leptospiral serovars (serovars australis, autumnalis, ballum, bataviae, canicola, grippotyphosa, hardjoprajitno, hebdomadis, icterohaemorrhagiae, javanica, pomona, pyrogenes, panama, and tarassovi) was carried out by using a pair of primers . Each serovar had a unique and distinct fingerprint pattern . DNAs of other bacterial species, including Escherichia coli, Pasteurella multocida, Salmonella spp., Pseudomonas spp., and Klebsiella spp., did not show any amplification . RAPD fingerprinting was found to be a rapid and sensitive method for serovar identification when it was compared to DNA restriction enzyme analysis, which produced a larger number of bands that made it more difficult to compare serovars. Curr Microbiol, 1997 Apr, 34(4), 244 - 9 Serotyping and enzyme characterization of Pasteurella haemolytica and Pasteurella multocida isolates recovered from pneumonic lungs of stressed feeder calves; Purdy CW et al.; Ninety-one isolates of Pasteurella multocida (Pm)and 124 of Pasteurella haemolytica (Ph) were recovered from the lungs of calves that died of bovine respiratory tract disease (BRTD) . Nine Pm enzyme profiles (A through I) and 9 Ph enzyme profiles (J through R) were determined for the Pasteurella isolates . The Pm isolates were relatively evenly divided among the enzyme profiles, with one exception, profile I . The Ph isolates were not evenly distributed among the profiles . Fifty of the 91 Pm isolates were serotyped . Forty-two Pm isolates were positive for capsule type A, and 8 were untypable . Five somatic type antigen profiles (3; 3,4; 3,7; 3,4,7; and 4) were identified among the 50 serotyped Pm isolates; one isolate was untypable . The Ph isolates were further divided through serotyping and grouped as follows: 74 (60%) Pasteurella haemolytica A1 (PhA1), 12 (10%) PhA2, 4 (3%) PhA5, and 34(27%) PhA6 . Eighty-one percent of the Ph serotypes were clustered in the M and N enzyme profile . The P enzyme profile was almost unique to PhA2 (8 of 12, 67% of PhA2 isolates) . Results of this study indicate a need to collect more data on Ph serotypes at the state veterinary diagnostic laboratories. J Med Chem, 1997 Mar 14, 40(6), 1041 - 5 Repromicin derivatives with potent antibacterial activity against Pasteurella multocida; McFarland JW et al.; Reductive amination of repromicin with polyfunctional amines has led to new macrolide antibacterial agents, some of which are highly potent against the Gram-negative pathogen Pasteurella multocida both in vitro and in a mouse infection model . A key element in this discovery was the recognition that among certain known macrolides increasing lipophilicity results in diminished in vivo activity . One repromicin derivative, 20-{N-{3-(dimethylamino)-propyl}-N-L-alanylamino}-20-deoxorepro micin (35), was selected for advanced evaluation . At 5 mg/kg, a single subcutaneous dose was found to control induced pasteurellosis in swine and induced respiratory disease in cattle. Vet Microbiol, 1997 Mar, 54(3-4), 369 - 74 RSF1010-based shuttle vectors for cloning and expression in Pasteurella multocida; Lee MD et al.; The broad host-range cloning vectors, pJRD215 and pMMB67EH, were evaluated for stability and cloning efficiency in Pasteurella multocida . Transformation of P . multocida by electroporation was unreliable and poorly efficient regardless of whether the transforming DNA was isolated from E . coli or P . multocida . Both vectors contain a mob site that enabled transfer by conjugation from E . coli to P . multocida with high efficiency . Kanamycin, streptomycin, and ampicillin resistance encoded by the vectors were expressed in P . multocida . LacZ was cloned in pMMB67EH, an expression vector, and was transferred to P . multocida by conjugation . The transconjugants expressed a functional beta-galactosidase as determined by o-nitrophenyl-beta-D-galactopyranoside (ONPG) test . We propose the use of these cosmid and expression vectors as a shuttle vectors for cloning in P . multocida. Vet Microbiol, 1997 Mar, 54(3-4), 343 - 55 Pasteurella multocida enters polarized epithelial cells by interacting with host F-actin; Rabier MJ et al.; We investigated the interaction of an avian strain of Pasteurella multocida with the cytoskeleton of MDCK cells, which formed a polarized epithelium when grown on type I collagen coated filters . Bacteria were incubated with MDCK cells for 30 min . 2, 4 and 6 hours and their location and association with the cell cytoskeleton determined by double-label immunofluorescence confocal microscopy . Cells were stained with a polyclonal antiserum to the outer-membrane proteins of P . multocida and with rhodamine phalloidin which specifically binds filamentous (F) actin . Confocal microscopy revealed that bacteria entered the cells by 30 min, and that by 6 hours there was a marked alteration in the actin cytoskeleton in which long filaments were reorganized to discrete foci of short actin filaments, within which were one or more bacteria . Electron microscopy demonstrated that by 2 hours, each bacterium was associated with many short 5-6 nm filaments . Treatment of MDCK cells with cytochalasin D for either 30 minutes or 24 hours prior to infection disrupted the actin cytoskeleton and inhibited entry of P . multocida. Semin Respir Infect, 1997 Mar, 12(1), 54 - 6 Pasteurella multocida pneumonia; Klein NC et al.; Pasteurella multocida, a gram-negative coccobacillus which colonizes the nasopharynx and gastrointestinal tract of many animals, is a well known cause of soft tissue infection after animal bites . Human infection can also occur after non-bite animal exposure, usually via inhalation of contaminated secretions . The respiratory tract is the second most common site of Pasteurella infection after soft tissue infection . Most patients with Pasteurella pulmonary infection are elderly with underlying lung disease, either COPD, bronchiectasis, or malignancy . The spectrum of disease includes pneumonia, tracheobronchitis, lung abscess, and empyema . Clinical features of Pasteurella respiratory tract infections are indistinguishable from other pathogens . A history of cat or dog exposure should alert the clinician to consider Pasteurella as a potential pulmonary pathogen in an elderly patient with chronic lung disease . The preferred drug for the treatment of Pasteurella infections is penicillin . Alternately, doxycycline is highly effective against P multocida. Appl Environ Microbiol, 1997 Mar, 63(3), 924 - 30 Nitrate-reducing bacteria on rat tongues; Li H et al.; Nitrite-producing bacteria (NPB) were isolated from tongues of laboratory rats . The most commonly found nitrite-producing organism was Staphylococcus sciuri, followed by Staphylococcus intermedius, Pasteurella spp., and finally Streptococcus spp . Both morphometric quantification of bacteria on tongue sections and enumeration of culturable bacteria (CFU) showed an increase in the density of bacteria towards the posterior tongue . Up to 65% of bacteria were located in the deep clefts on the posterior tongue . The proportion of culturable NPB in the total culturable microbial population increased from 6% (10(5) CFU cm-2) on the anterior tongue to 65% (10(7) CFU cm-2) on the posterior tongue . Different species compositions of NPB were found on different tongue sections with S . intermedius populations decreasing and S . sciuri and Pasteurella populations increasing towards the posterior tongue . Nitrite production was sensitive to oxygen, and significant nitrite production was only detected on the posterior tongue where the majority of bacteria are situated in deep clefts in the tongue surface . This study suggests the importance of bacteria in nitrite production, from nitrate, on the tongue . Nitrite produced on the tongue may subsequently form nitric oxide in the acidic environment of the stomach . Because of the antimicrobial properties of nitric oxide, a key role for nitrate-reducing tongue bacteria in host animal defense against food-borne pathogens in proposed. Infect Immun, 1997 Mar, 65(3), 957 - 63 Increase of glycocalyx and altered lectin agglutination profiles of Pasteurella haemolytica A1 after incubation in bovine subcutaneous tissue chambers in vivo or in ruminant serum in vitro; Brogden K et al.; Pasteurella haemolytica serotype A1 (bovine strain OK) was incubated for 2 and 6 h in bovine subcutaneous tissue chambers in vivo, and ovine strain 82-25 and bovine strain L011 were incubated in vitro for 2 h in heat-inactivated ovine or bovine serum from which gamma globulin had been depleted by protein G affinity chromatography to assess changes in morphology and lectin agglutination profiles (strains 82-25 and L101 only) . Cells, removed from chambers after 2 h, were covered with an extensive, dense glycocalyx extending approximately 0.5 microm from the cell surface . In many cells, the glycocalyx was separated from the cell surface by a clear, electron-transparent area . Cells, removed at 6 h, were covered with a sparse glycocalyx of fine fibers 0.2 to 0.3 microm from the cell surface . Strains 82-25 and L101, incubated for 2 h in heat-inactivated ovine or bovine serum or in heat-inactivated ovine or bovine serum depleted of gamma globulin by protein G affinity chromatography, were also covered with a glycocalyx . The glycocalyx did not bind protein A-colloidal gold and therefore did not contain aggregates of accumulated antibody . Strains 82-25 and L101 were incubated individually for 2 h in 10 mM sodium phosphate buffer (pH 7.2) containing 0.14 M NaCl, 0.5 mM CaCl2, and 0.15 mM MgCl2 or with this buffer and either 25% heat-inactivated, gamma globulin-depleted ovine serum or 25% heat-inactivated, gamma globulin-depleted bovine serum . Agglutination profiles were then determined with 17 lectins in 10 mM HEPES-buffered saline (pH 8.4) with 0.1 mM CaCl2 and 0.08% sodium azide . Profiles did not vary with 10 of 17 lectins . However, profiles did vary with peanut agglutinin, Phaseolus vulgaris leucoagglutinin, Sophora japonica agglutinin, Maackia amurensis lectin II, Narcissus pseudonarcissus (daffodil) lectin, Griffonia simplicifolia lectin I, and Pisum sativum agglutinin . Altered profiles indicate a change in the bacterial cell surface, possibly by adsorption or alteration of surface carbohydrate moieties by serum constituents. Gene, 1997 Feb 28, 186(2), 207 - 11 Plasmids for heterologous expression in Pasteurella haemolytica; Fedorova ND et al.; New cloning and expression vectors that replicate both in Pasteurella haemolytica and in Escherichia coli were constructed based on a native sulfonamide (SuR) and streptomycin (SmR) resistant plasmid of P . haemolytica called pYFC1 . Each shuttle vector includes an MCS and a selectable antibiotic resistance marker that is expressed in both organisms . Plasmid pNF2176 carries the P . haemolytica ROB-1 beta-lactamase gene (blaP, ApR) and pNF2214 carries the Tn903 aph3 kanamycin resistance (KmR) element . The expression vector, pNF2176, was created by placing the MCS downstream of the sulfonamide gene promoter (PsulII) on pYFC1; this was used to clone and express the promoterless Tn9 chloramphenicol resistance gene (cat, CmR) in P . haemolytica (pNF2200) . A promoter-probe vector (pNF2283) was constructed from pNF2200 by deleting PsulII. Zentralbl Bakteriol, 1997 Feb, 285(3), 440 - 4 Pasteurella caballi infection following a horse bite; Escande F et al.; The isolation of Pasteurella caballi from an horse-bite wound in a 56-year-old man is reported . Biochemical characteristics are described and compared with the other species representing the genus Pasteurella . This strain probably represents the first human isolate of P . caballi in France. Vaccine, 1997 Feb, 15(2), 203 - 8 Cross protective immunity conferred by a marker-free aroA mutant of Pasteurella multocida; Homchampa P et al.; The aroA gene from Pasteurella multocida serotype A:1 (X-73) was cloned by complementation of the Escherichia coli aroA mutant AB2829 with a DNA library constructed in pUC18 . The cloned aroA gene was inactivated by deletion of a 300 bp internal sequence and reintroduced by homologous recombination into the chromosome of X-73 and P-1059 (serotype A:3) using a Pasteurella-E . coli shuttle vector pPBA1100 . By subjecting the transformed cells to repeated subculturing in the presence of antibiotic selection coupled with auxotrophic enrichment, marker-free aroA mutants of X-73 and of P-1059 were isolated and designated PMP1 and PMP3, respectively . PMP1 and PMP3 were highly attenuated and capable of conferring complete protection against subsequent lethal challenge infection in a mouse model . Moreover, PMP3-immunized mice were protected against heterologous challenge infection with serotype A:1 or A:4. J Small Anim Pract, 1997 Feb, 38(2), 51 - 6 New combination for the therapy of canine otitis externa . I . Microbiology of otitis externa; Kiss G et al.; In order to compound a new drug combination against canine otitis externa (OE), 515 dogs affected with OE were subjected to physical examination and microbiological analysis of their ear exudates . OE was erythematous-ceruminous in 83 per cent and suppurative in 17 per cent of the patient material . Erythematous-ceruminous inflammations were characterised by severe pruritus and accumulation of brownish, greasy cerumen in the auditory canal . The yeast Malassezia pachydermatis was isolated from the ears of 76 per cent of the dogs, often in combination with Staphylococcus intermedius bacteria . M pachydermatis showed the most sensitivity, in decreasing order of efficacy, to ketoconazole, econazole, clotrimazole, miconazole and nystatin . S intermedius isolates were most sensitive to amoxycillin-clavulanic acid, enrofloxacin, cephalexin and gentamicin . The microorganism most frequently isolated from dogs with suppurative OE was Pseudomonas aeruginosa; in some cases Proteus, Streptococcus and Pasteurella were also isolated . The P aeruginosa isolates showed the highest sensitivity to gentamicin, polymyxin B and tobramycin. Vet Microbiol, 1997 Feb, 54(2), 167 - 83 Outer membrane proteins of bovine Pasteurella multocida serogroup A isolates; Dabo SM et al.; The outer membrane proteins (OMPs) of P . multocida serotypes A3 (7 isolates), A4 (2 isolates), A3,4 and A2 (one isolate each) obtained from pneumonic cattle (10 isolates) and from one pig isolate were investigated to identify potential immunogens . SDS-PAGE of P . multocida OM isolated by SDG centrifugation of spheroplasts revealed eight major OMPs . Outer membranes isolated by sarcosyl extraction or SDG had similar protein composition on Coomassie blue-stained SDS-PA gel and on immunoblots . Two major OMPs (M(r)s of 35 and 46 kDa at 100 degrees C) demonstrated heat modifiability with apparent M(r)s of 30 and 34 kDa at 37 degrees C, respectively . The N-terminal aa sequences of these heat modifiable proteins revealed homology with E . coli OmpA and Hib P1 proteins, respectively . Protease treatment of whole cells followed by western immunoblots using bovine convalescent sera identified several immunogenic, surface-exposed and conserved OMPs among the eleven P . multocida i