Pasteurella

Vet Immunol Immunopathol, 1995 Nov, 49(1-2), 61 - 74
Characterisation of bovine lipopolysaccharide binding protein and the in vivo acute phase response to Pasteurella haemolytica Type A; Horadagoda NU et al.; Lipopolysaccharide binding protein (LBP) is a liver-derived acute phase protein which is implicated in modulating the host responses to lipopolysaccharide (LPS) from Gram-negative bacteria . LBP interacts with circulatory LPS to form complexes which bind to the CD14 receptor or cells of the monocytic lineage and neutrophils resulting in their activation . This causes the release of mediators and cytokines which are responsible for initiating the acute phase response . LBP-like activity has now been identified in bovine serum and in this study LBP has been purified from acute phase bovine serum using ion exchange chromatography . On sodium dodecyl sulphate polyacrylamide electrophoresis, bovine LBP demonstrated a single band with a molecular mass of 58 kDa . Bovine LBP enhanced the binding of LPS to human monocytes while enzymatic removal of the CD14 receptor abrogated this interaction . Furthermore, bovine LBP increased the sensitivity of monocytes to LPS by at least 100-fold . Depletion of LBP by means of antibodies to bovine LBP inhibited the serum mediated LPS binding to monocytes . Antibodies to rabbit LBP or recombinant human LBP did not cross-react with bovine LBP . Studies on the kinetics of LBP activity in calves during the acute phase response demonstrated a four-fold increase in the serum concentration 36 h after a single intratracheal inoculation of Pasteurella haemolytica A1 . The findings of this study indicate that cattle possess a LPS detection mechanism comparable to that described in man and experimental animals in which LBP forms complexes in serum with circulatory LPS enhancing the signal to the immune system to mount a host response . The isolation of LBP will allow further investigations into LBP-mediated responses to LPS in cattle.

Vet Immunol Immunopathol, 1995 Nov, 49(1-2), 15 - 28
Increased tumor necrosis factor-alpha and interleukin-1 beta expression in the lungs of calves with experimental pneumonic pasteurellosis; Yoo HS et al.; We used a well characterized pneumonic pasteurellosis model in calves to determine whether increased proinflammatory cytokines, tumor necrosis factor-alpha (TNF-alpha) and interleukin-1 beta (IL-1 beta) expression and secretion were associated with pneumonic lesions . Bronchoalveolar lavage fluids, lavage cells consisting of alveolar macrophages and neutrophils with degenerative changes, and lung tissues were analyzed for the presence of TNF-alpha and IL-1 beta approximately 48 h following endobronchial inoculation of logarithmic phase Pasteurella haemolytica 12296 organisms . Levels of TNF-alpha and IL-1 beta mRNA were significantly increased in lavage cells of P . haemolytica-infected animals but not in cells from phosphate buffered saline (PBS) inoculated controls based on in situ hybridization analysis . Significantly increased levels of TNF-alpha, and IL-1 beta mRNA were also expressed within the pneumonic lesions from P . haemolytica-infected calves . In contrast, lung tissues from PBS-inoculated control calves had cytokine mRNAs expressed at extremely low levels . Increased levels of bioactive IL-1 and immunoreactive (not bioactive) TNF-alpha were found in lavage fluids from P . haemolytica-infected calves compared with lavage fluids from PBS-inoculated calves . These findings indicate that the proinflammatory cytokines TNF-alpha and IL-1, may be associated with pathogenesis of lung injury in bovine pneumonic pasteurellosis.

Vet Immunol Immunopathol, 1995 Nov, 49(1-2), 143 - 59
Occurrence of T lymphocytes in perivascular regions of the lung after intratracheal infection of swine with Pasteurella multocida; Berndt A et al.; The defence mechanisms of the lung acting against airborne antigens (bacteria) are not yet fully understood . Therefore, we investigated cell-derived immunological processes in porcine pulmonary tissue . The incidence and distribution of leukocyte subpopulations in lungs, lung lymph nodes and tonsils from eight animals intratracheally infected with Pasteurella multocida (P.m.) type A wild strain and from four non-infected control animals were established by immunohistochemistry, using a panel of monoclonal antibodies (mAbs) against specific porcine leukocyte antigens: MHC Class II (MHC II), SWC1, SWC3a, CD2a, CD4a, CD8b . In the test animals, a broad layer of SWC1+ cells and SWC3a+ cells emerged in the subsinusoidal region of lung lymph nodes and in the bronchoalveolar spaces of the lung as early as 4 and 24 h after infection, and in the subsinusoidal regions of the lung lymph nodes only very few cells could be stained with the mAb against the MHC II antigen . We could not detect any MHC Class II+ cells in the bronchoalveolar spaces at this time of the investigation . In the course of the disappearance of the SWC1+ cells and SWC3+ cells, CD2a+, CD4a+ and, in lower numbers, CD8b+ T lymphocytes seemed to concentrate in the perivascular and, partially, the peribronchial regions of the lung 72 h after infection . By means of the double immunostaining method, we could demonstrate an accumulation of CD4a+ cells and CD8b+ cells after infection which lacked the SWC1 antigen, indicating that these cells were activated T lymphocytes . The same cell types (SWC1-/CD4a+ and SWC1-/CD8b+ cells) as well as CD8b-/CD4a+ cells could be observed in the interstitium of the lung 72 h after infection.

Res Vet Sci, 1995 Nov, 59(3), 267 - 71
Evaluation of the nebulisation of sodium ceftiofur in the treatment of experimental Pasteurella haemolytica bronchopneumonia in calves; Sustronck B et al.; Severe acute bronchopneumonia was induced in 24 conventional Friesian Holstein calves by inoculating them intratracheally with Pasteurella haemolytica type A1 . Twelve of the calves were treated intramuscularly with sodium ceftiofur and 12 were treated with an aerosol of sodium ceftiofur . The mortality rate in the group of calves treated with the aerosol was significantly lower, and their clinical and haematological parameters returned to normal significantly faster than in the calves treated intramuscularly.

J Antibiot (Tokyo), 1995 Nov, 48(11), 1330 - 5
Derivatives of tetrodecamycin; Tsuchida T et al.; The derivatives of tetrodecamycin (1), being introduced acyl, carbamoyl and alkyl groups at 14-hydroxyl group and modified at exo-methylene group, were synthesized and evaluated on their antibacterial activities . Although 14-O-substituted tetrodecamycins (3 approximately 19) showed weak activity against Pasteurella piscicida, they were more active against Gram-positive bacteria than 1 . Among them, 15 showed approximately 10-fold higher activity than 1 . The derivatives (20 approximately 23) modified at 4 or 5 positions had moderate antibacterial activity . The absolute structure of 4(R),5-dibromotetrodecamycin (23) was determined by X-ray crystallographic analysis.

J Leukoc Biol, 1995 Nov, 58(5), 510 - 8
Evidence for a third component in neutrophil aggregation: potential roles of O-linked glycoproteins as L-selectin counter-structures; Bennett TA et al.; The homotypic aggregation of neutrophils requires the participation of L-selectin and the beta 2-integrins, but it has not been clear whether the two receptors recognize one another as counter-structures or whether other adhesion molecules are involved . We have examined aggregation of live neutrophils with target populations, manipulated to alter expression of adhesive epitopes, using flow cytometry . A target population depleted of both L-selectin and activatable beta 2-integrin displayed an ability to aggregate with live neutrophils, suggesting that these two molecules are not counter-structures . We also found that an O-sialoglycoprotease (GCP) from Pasteurella haemolytica is capable of inhibiting homotypic aggregation . Neutrophils treated with GCP lose O-glycosylated proteins but retain L-selectin and activatable beta 2-integrin . One or more of the GCP substrates appears to function in L-selectin-dependent binding but not in beta 2-integrin-dependent binding . Together the data suggest a mechanism of aggregation that is analogous to leukocyte-endothelial cell adhesion in which a low-affinity carbohydrate-dependent interaction precedes a high-affinity integrin-dependent adhesion.

Curr Microbiol, 1995 Nov, 31(5), 312 - 5
Extracellular neuraminidase production by Pasteurella species isolated from infected animals; Straus DC et al.; A total of 721 field isolates of various Pasteurella species (haemolytica, multocida, and testudinis) from various regions of the United States were examined for extracellular neuraminidase production . All strains were grown and tested in the same way . Included were 372 P . haemolytica serotype 1 isolates, 181 P . haemolytica serotype 2 isolates, 63 P . haemolytica serotype 6 isolates, 101 Pasteurella multocida isolates, and 4 Pasteurella testudinis isolates . All Pasteurella species examined produced the enzyme . The data revealed the following: (1) Several transfers of P . haemolytica strains on blood agar medium did not cause a decrease in enzyme activity . (2) P . haemolytica serotypes 2 and 6 produce more neuraminidase than P . haemolytica serotype 1, P . multocida, and P . testudinis isolates . (3) There was no apparent change in neuraminidase production by P . haemolytica serotypes 1 and 2 obtained from the same animal taken on different days in the feedyard . (4) There was no significant change in neuraminidase production by P . haemolytica serotype 2 isolates taken from the same animal at the auction market and later at the feedyard.

FEMS Microbiol Lett, 1995 Oct 15, 132(3), 247 - 51
Pili of Pasteurella multocida of porcine origin; Isaacson RE et al.; Using electron microscopy, pili with at least two distinct morphologies were observed on strains of Pasteurella multocida isolated from pigs with atrophic rhinitis . Rigid pili were found on 60-80% of all cells observed . These pili had a strong tendency to lie flat along the side of the outer cell membrane of P . multocida and as a result frequently were difficult to see . After growth in vitro, piliated P . multocida cells produced few pili (approx . 3-5 per cell) . Heavily piliated cells were occasionally observed . The second type of pili were curly and also were difficult to visualize . Cells from cultures containing piliated cells failed to attach to red blood cells and to immobilized mucus.

Am J Vet Res, 1995 Oct, 56(10), 1317 - 21
Passive protection of calves with Pasteurella haemolytica antiserum; Mosier DA et al.; Four colostrum-deprived calves each were immunized passively with antisera to whole Pasteurella haemolytica, leukotoxin-containing supernatants of P haemolytica, P haemolytica lipopolysaccharide, or newborn calf serum . Calves were challenge exposed intrabronchially with 5 x 10(9) P haemolytica, and 24 hours later, the resulting lesions were evaluated . The greatest protection against challenge exposure was provided by the antiserum to whole P haemolytica (lesion score = 6.3), whereas newborn calf serum provided the least protection (lesion score = 28.3) . Calves that received antiserum to P haemolytica supernatants were moderately protected (lesion score = 16.3), and the antiserum to lipopolysaccharide provided minimal protection (lesion score = 21.8) . Antibodies that were unique to whole P haemolytica antiserum and produced dense bands on immunoblots were detected to antigens at 66, 50, and 30 kd . Antibodies in the supernatant preparation that produced prominent bands reacted to antigens between 100 and 90 kd . Collectively, antibodies to these antigens may be responsible for enhancing resistance to experimentally induced pneumonic pasteurellosis . Antibodies to antigens in P haemolytica lipopolysaccharide provided little to no protection.

Rev Latinoam Microbiol, 1995 Oct-Dec, 37(4), 353 - 65
{Pathogenesis of lung damage caused by Pasteurella haemolytica}; Ramirez-Romero R et al.; Pneumonic pasteurellosis is the major economic problem of the cattle industry in North America . This disease is characterized by an acute, severe, fibrinonecrotic pleuropneumonia . Pasteurella haemolytica A1 is commonly isolated from these pneumonic lesions . It has been demonstrated that stress or viral infection compromises defense mechanisms of the upper respiratory tract and lung, predisposing to an initial multiplication of bacteria in the nasopharynx and, subsequently, lungs are deluged with large numbers of bacteria . Once multiplication in the alveoli has begun, virulence factors exert their influence to induce an excessive host inflammatory response that results in severe tissue damage . Despite a large number of studies conducted to explore the complex interaction between P . haemolytica and the host response, there still remains a lack of detailed understanding . This review discusses evidence of the role of the main virulence factors of P . haemolytica on the pathogenesis of pulmonary damage.

Zentralbl Veterinarmed A, 1995 Oct, 42(8), 531 - 44
Pulmonary ventilation, mechanics, gas exchange and haemodynamics in calves following intratracheal inoculation of Pasteurella haemolytica; Linden A et al.; A Pasteurella haemolytica A1 broth was injected intratracheally in eight calves and measurements of pulmonary function values (PFV) were made once before and hourly post inoculation (p.i.) . Changes in PFVs, included increased respiratory rate and minute ventilation (up to 158% of baseline 2 h p.i.) and decreased tidal volume and lung dynamic compliance (up to 33% of baseline 3 h p.i.) . Total pulmonary resistance was not affected . At and after 3 h p.i . there was a progressive impairement of gas exchange, as judged from arterial O2 tension which decreased up to 65% of baseline . In contrast, arterial CO2 tension was not affected . Pulmonary hypertension was observed during the 3 last h of the study and was attributable to an increased pulmonary vascular resistance . Severe neutropenia was observed at 3 h p.i . and post-mortem histological findings were consistent with an acute fibrinohemorragic bronchopneumonia . In conclusion, P . haemolytica airway challenge unequiovocally resulted in acute pneumonia, providing a reproducible pathophysiological model for investigations regarding new therapeutic strategies.

Lab Anim Sci, 1995 Oct, 45(5), 526 - 32
Protective immunity to Pasteurella multocida heat-labile toxin by intranasal immunization in rabbits; Suckow MA et al.; Heat-labile Pasteurella multocida toxin (PMT) is an important virulence factor of some isolates from rabbits . To determine whether protective immunity to PMT could be induced in rabbits by intranasal immunization with heat-inactivated PMT, we immunized groups of rabbits intranasally at days 0, 7, 14, and 21 with inactivated PMT, with or without cholera toxin, an adjuvant for the mucosal immune system . Significant increases in anti-PMT IgA in nasal lavage samples and anti-PMT serum IgG, as determined by enzyme-linked immunosorbent assay, developed within 2 weeks after initial immunization . Coadministration of cholera toxin with inactivated PMT enhanced anti-PMT activity in the samples . Rabbits similarly immunized on days 0, 7, and 14 were challenged with PMT, and tissues were graded histologically on a numeric scale of lesion severity . Immunization conferred partial protection against development of pneumonia, pleuritis, hepatic necrosis, and testicular atrophy in rabbits challenged 16 days after initial immunization . Thus, immunization with inactivated PMT stimulates a protective response to PMT challenge in rabbits that is enhanced by coadministration of cholera toxin.

Vet Microbiol, 1995 Oct, 46(4), 401 - 13
Stimulation of avian respiratory phagocytes by Pasteurella multocida: effects of the route of exposure, bacterial dosage and strain, and the age of chickens; Hassanin HH et al.; The effects of the route of exposure (intratracheal {IT}, drinking water {DW} and aerosolization {AS}), the age of chickens, and the dose of two vaccine strains of Pasteurella multocida (CU and M-9) on the number, phagocytic proportion and capacity of macrophages, granulocytes and lymphocytes (collectively avian respiratory phagocytes {ARPs}) were analyzed . Administration of P . multocida via the DW even at very high dose failed to stimulate ARPs . In contrast, administration of both strains of P . multocida either IT or by AS resulted in rapid and highly significant increases in the numbers of ARPs . 10 x 10(9) colony forming units (cfu) of aerosolized P . multocidaCU strain activated ARPs maximally in both young (3-6 weeks of age) and old (4-6 months of age) chickens . Old chickens responded in dose dependent manner to 20 x 10(9), 8 x 10(9), and 4 x 10(9) cfu of aerosolized P . multocida CU strain . Young chickens responded significantly only to 8 x 10(9) CU organisms . The M-9 and CU strains had limited differences in inducing migration of ARPs into the respiratory system of chickens or elevating phagocytic proportions and capacity of ARPs . The results indicate that the analyzed factors influence the response of ARPs to P . multocida to various degrees.

Vet Microbiol, 1995 Oct, 46(4), 393 - 400
Bronchopneumonia in mice caused by Pasteurella haemolytica A2 after predisposition by ovine Bordetella parapertussis; Porter JF et al.; Initial intranasal inoculation of four to eight-week-old Swiss White mice with 7.5 x 10(6) colony forming units (cfu) of ovine B . parapertussis followed 30 min, three or five days, by intranasal inoculation with 1.4 x 10(5) cfu of Pasteurella haemolytica A2 resulted in a more severe infection pattern than when either agent was administered alone . Histopathological examination showed that inoculation with B . parapertussis alone caused a bronchopneumonia the severity of which was dependant upon the infecting dose . Bacteria were recovered up to 10 days after inoculation . P . haemolytica alone had no apparant pathogenic effect and was cleared from the lungs within 24 h . When both agents were given in combination the lesions were most severe when P . haemolytica was administered three days after B . parapertussis infection . These findings suggest that B . parapertussis predisposes mice to subsequent infection with P . haemolytica and that the timing of the P . haemolytica administration influences the severity of the lung lesions.

Can J Vet Res, 1995 Oct, 59(4), 311 - 5
The influence of supplemental chromium and vaccines on the acute phase response of newly arrived feeder calves; Wright AJ et al.; The acute phase response as indicated by serum haptoglobin and total haemolytic complement activity (CH50) was measured in 72 cross-bred steer calves purchased at sales in Ontario . During the 28 day (d) trial, 18 steers were randomly assigned to each of the following groups: 1) control; 2) vaccinated (Infectious Bovine Rhinotracheitis, Parainfluenza-3, Bovine Viral Diarrhea, Bovine Respiratory Synctial Virus vaccine plus Pasteurella haemolytica vaccine); 3) supplemental chelated Cr (0.14 mg/kg); and 4) Cr plus vaccines . Haptoglobin concentrations were low at arrival, increased (P < 0.05) on day 7, and returned to near initial levels (P > 0.05) by day 14 . Supplemental Cr reduced (P < 0.05) haptoglobin on day 7 when morbidity was highest . Following antibiotic treatment for respiratory disease haptoglobin was lower (P < 0.05) than during morbidity; however, during morbidity, haptoglobin concentrations were not greater in sick calves (P > 0.05) than in healthy calves . Complement activity was lowest on day 7 (P < 0.05) and peaked on day 14 (P < 0.05) . Complement activity tended to be lower on day 7 for vaccine, Cr, and Cr+ vaccine groups; however, the difference from controls was not significant (P > 0.10) . Complement activity did not increase on day 14 (P > 0.05) with Cr supplementation as in other treatments . Morbid calves had lower (P < 0.05) CH50 activity than healthy calves on day 14 . Following antibiotic treatment, the Cr-supplemented group had higher (P < 0.05) CH50 than during morbidity . In general, chromium supplementation reduced the acute phase response in newly arrived feeder calves.

Cancer Res, 1995 Oct 1, 55(19), 4425 - 31
Differential colon cancer cell adhesion to E-, P-, and L-selectin: role of mucin-type glycoproteins; Mannori G et al.; E-, P-, and L-selectin support the adhesion of leukocytes to the vessel wall through the recognition of specific carbohydrate ligands, which often contain sialylated, fucosylated lactosamines such as sialyl Lewis x {sLex; Neu5Ac alpha 2-3Gal beta 1-4(Fuc alpha 1-3)GlcNAc-} . E-selectin expressed by activated endothelium has been shown to support the adhesion of sLex-bearing colon cancer cells . In the present study, we examine the interactions of multiple colon cancer cell lines with all three selectins . Three colon cancer cell lines (LS 180, T84, and COLO 205) bound to recombinant purified E-, P-, and L-selectin . The colon cancer line COLO 320 bound to P- and L-selectin but not E-selectin; conversely, HT-29 cells bound E-selectin but not P- and L-selectin . Caco-2 showed little or no interaction with any of the three selectins . Treatment of the cells with O-sialoglycoprotease from Pasteurella haemolytica, an enzyme that selectively cleaves mucin-type O-linked glycoproteins, reduced binding to purified P- and L-selectin in all cases . In addition, recombinant soluble P- and L-selectin bound to affinity-purified mucins from all adherent tumor cell lines . Of the four tumor cell lines that interacted with E-selectin, O-glycoprotease treatment substantially diminished adhesion of LS 180 and T84, had little effect on COLO 205, and failed to inhibit the binding of HT-29 . As predicted by these data, E-selectin showed substantial binding only to mucins purified from LS 180 and T84 . These findings suggest that L- and P-selectin interact primarily with mucin-type ligands on colon cancers, whereas E-selectin can recognize both mucin and nonmucin ligands . Binding of the colon cancer lines to purified selectins correlates with their adhesion to activated endothelial cells (E-selectin-dependent), platelets (P-selectin-dependent), and neutrophils (L-selectin-dependent) . These differential tumor cell-selectin interactions may influence metastatic spread and may also contribute to the observed variability in host response to tumor progression.

J Antibiot (Tokyo), 1995 Oct, 48(10), 1110 - 4
Tetrodecamycin and dihydrotetrodecamycin, new antimicrobial antibiotics against Pasteurella piscicida produced by Streptomyces nashvillensis MJ885-mF8 . II . Structure determination; Tsuchida T et al.; Novel antimicrobial antibiotics against Pasteurella piscicida, tetrodecamycin (1) and weakly active dihydrotetrodecamycin (2) were isolated from a culture broth of Streptomyces nashvillensis MJ885-mF8 . The planar structure of 1 was determined to be 2-acyl-4-ylidene tetronic acid alkyl ether containing decaline ring by various NMR spectral data of 1 and its acetyl derivative (3) . The structure of 2 was elucidated by comparison with the spectral data of 1 and confirmed by catalytic reduction of 1 into 2 . The X-ray crystallography of 2 showed the relative stereochemistry . Their absolute configurations were determined by using modified Mosher's method.

J Antibiot (Tokyo), 1995 Oct, 48(10), 1104 - 9
Tetrodecamycin and dihydrotetrodecamycin, new antimicrobial antibiotics against Pasteurella piscicida produced by Streptomyces nashvillensis MJ885-mF8 . I . Taxonomy, fermentation, isolation, characterization and biological activities; Tsuchida T et al.; The novel antimicrobial antibiotic against Pasteurella piscicida, tetrodecamycin (1) and weakly active dihydrotetrodecamycin (2) were isolated from the fermentation broth of Streptomyces nashvillensis MJ885-mF8 . They were purified by adsorption on Diaion HP-20, silica gel column chromatography and crystallization . The MICs of 1 were 6.25 approximately 12.5 micrograms/liter and 1.56 approximately 6.25 micrograms/ml against Gram-positive bacteria including methicillin-resistant Staphylococcus aureus (MRSA) and 12 strains of P . piscicida, respectively.

J Infect, 1995 Sep, 31(2), 161 - 2
Meningitis in infancy caused by Pasteurella multocida; Boocock GR et al.; A high proportion of household pets are colonised by Pasteurella multocida . The organism can be transmitted to humans by contact with animal saliva and is a recognised, although rare, cause of meningitis in infancy . Intimate contact between infants and family pets should be discouraged.

Vet Microbiol, 1995 Sep, 46(1-3), 335 - 42
Molecular studies on avian strains of Pasteurella multocida in Australia; Diallo IS et al.; A collection of 45 strains of Pasteurella multocida was assembled . The strains had been isolated from cases of fowl cholera in eastern Australia over 8 years, and included mainly type A strains . All the strains were examined for plasmids and resistance to 10 antimicrobial agents and most of the strains were examined for restriction fragment length polymorphism . Nine strains were assayed for pathogenicity for mice . Twenty strains yielded no plasmid . Seven contained a single plasmid of 1.3 kbp and 18 contained 2 plasmids, of 2.4 and 7.5 kbp . All the strains were resistant to streptomycin, trimethoprim and lincomycin while one strain was resistant to tetracycline . There was no correlation between plasmid content and resistance to antimicrobial agents . Three strains that lacked plasmids were highly virulent for mice, 6 strains containing plasmids were not . Restriction fragment length polymorphism generated by HpaII allowed the 39 strains that were tested to be divided into 10 groups.

Thorax, 1995 Sep, 50(9), 1017 - 8
Chronic lung abscess due to Pasteurella multocida; Machiels P et al.; A case of chronic lung abscess due to Pasteurella multocida presenting as a solitary pulmonary mass with a computed tomographic appearance suggestive of malignancy is described.

J Emerg Med, 1995 Sep-Oct, 13(5), 623 - 7
Actinobacillus ureae meningitis: case report and review of the literature; Kingsland RC et al.; Actinobacillus urea, formerly known as Pasteurella ureae, is an uncommon commensal of the upper respiratory tract in humans . It has been identified as the primary pathogen in 10 cases of meningitis and several cases of pneumonia, sepsis, and peritonitis . A case is presented that represents another documented case of meningitis due to this rare organism . Risk factors associated with serious infection due to Actinobacillus ureae and basic management approaches to posttraumatic meningitis in general are discussed.

Infect Immun, 1995 Sep, 63(9), 3595 - 9
Pasteurella haemolytica lipopolysaccharide-associated protein induces pulmonary inflammation after bronchoscopic deposition in calves and sheep; Brogden KA et al.; The lipopolysaccharide (LPS)-associated protein (LAP) was extracted from Pasteurella haemolytica serotype A1 strains L101 (bovine origin) and 82-25 (ovine origin) . Extracts contained 0.017% total LPS and appeared as only two bands at 14 and 16.6 kDa after sodium dodecyl sulfate-polyacrylamide gel electrophoresis . To determine the extent of pulmonary inflammation induced by LAP and its possible role in the pathogenesis of pneumonic pasteurellosis, LAP (500 micrograms in pyrogen-free saline {PFS}) was deposited by fiber-optic bronchoscopy into the dorsum of the caudal portion of the cranial lobe of the right lung of calves (strain L101 LAP) and sheep (strain 82-25 LAP) . LPS (500 micrograms in PFS), 3-h P . haemolytica cultures (1.6 x 10(8) to 1.9 x 10(8) CFU in PFS), and PFS alone were deposited similarly as controls . At necropsy, 24 h after deposition, gross and histologic pulmonary lesions of calves and sheep given LAP, LPS, and P . haemolytica were similar and consisted of various degrees of acute bronchopneumonia (relative severities of lesions induced: LAP < LPS < live organisms) . By subjective histologic interpretation and semiquantitative morphometry, animals given LAP had the highest percentage of macrophages per alveolar lumen and the lowest percentage of neutrophils . The lesions from animals given LPS were more severe than those given LAP, but the morphometric cell counts were similar . In contrast, animals inoculated with P . haemolytica had lesions typical of this agent, consisting of many neutrophils, proteinaceous exudate, and a few macrophages . Morphometrically, these lesions had the highest numbers of neutrophils and the lowest numbers of macrophages . These studies show that LAP can induce an inflammatory response in the alveolar lumens and may play a role in the pathogenesis of pneumonic pasteurellosis.

Parasitology, 1995 Sep, 111 ( Pt 3), 247 - 55
Parasites of wild brown rats (Rattus norvegicus) on UK farms; Webster JP et al.; Wild brown rats (Rattus norvegicus) from 11 rural UK farmsteads were found to carry 13 zoonotic and 10 non-zoonotic parasitic species, many of which (e.g . Cryptosporidium, Pasteurella, Listeria, Yersinia, Coxiella and Hantavirus) have rarely or never been previously investigated for wild rats . The study suggests that wild brown rats, serving as vectors of disease, represent a serious risk to the health of humans and domestic animals in the UK.

Microbiology, 1995 Sep, 141 ( Pt 9), 2211 - 8
Functional analysis of the Bacillus subtilis purT gene encoding formate-dependent glycinamide ribonucleotide transformylase; Saxild HH et al.; The purT gene from Bacillus subtilis encoding the formate-dependent glycinamide ribonucleotide transformylase T was cloned by functional complementation of an Escherichia coli purN purT double mutant . The nucleotide sequence revealed an open reading frame of 384 amino acids . The purT amino acid sequence showed similarity to the enzyme phosphoribosylaminoimidazole carboxylase encoded by the purK gene but not to the N10-formyltetrahydrofolate-dependent glycinamide ribonucleotide transformylase N enzyme encoded by the purN gene . The glycinamide ribonucleotide transformylase T level was repressed in cells grown in rich medium compared to minimal-medium-grown cells . However, when the culture entered the stationary-growth phase the enzyme level increased in rich medium and decreased in minimal medium . By comparing the deduced amino acid sequence of the B . subtilis purT gene product with translated nucleotide sequences in various databanks, evidence for the existence of putative purT genes in the Gram-negative bacteria Pasteurella haemolytica and Pseudomonas aeruginosa was obtained.

J Clin Microbiol, 1995 Sep, 33(9), 2435 - 44
Comparison of MICs of ceftiofur and other antimicrobial agents against bacterial pathogens of swine from the United States, Canada, and Denmark; Salmon SA et al.; The MICs of ceftiofur and other antimicrobial agents, tested for comparison, for 515 bacterial isolates of pigs from the United States, Canada, and Denmark with various diseases were compared . The organisms tested included Actinobacillus pleuropneumoniae, Escherichia coli, Pasteurella multocida, Salmonella choleraesuis, Salmonella typhimurium, Streptococcus suis, Streptococcus dysgalactiae subsp . equisimilis, Streptococcus equi subsp . equi, and Streptococcus equi subsp . zooepidemicus . In addition to ceftiofur, the following antimicrobial agents or combinations were tested: enrofloxacin, ampicillin, sulfamethazine, trimethoprim-sulfadiazine (1:19), erythromycin, lincomycin, spectinomycin, lincomycin-spectinomycin (1:8), tilmicosin, and tetracycline . Tilmicosin was only tested against the U.S . isolates . Overall, ceftiofur and enrofloxacin were the most active antimicrobial agents tested against all isolates, with MICs inhibiting 90% of isolates tested (MIC90s) of < or = 2.0 and < or = 1.0 microgram/ml, respectively . Erythromycin, sulfamethazine, spectinomycin, and lincomycin demonstrated limited activity against all of the organisms tested, with MIC90s of > or = 8.0, > or = 256.0, > or = 32.0, and > or = 16.0 micrograms/ml, respectively . Trimethoprim-sulfadiazine was active against isolates of A . pleuropneumoniae, S . choleraesuis, S . typhimurium, P . multocida, S . equi, and S . suis (MIC90s, < or = 0.5 microgram/ml) but was less active against the E . coli strains tested (MIC90, > 16.0 micrograms/ml) . Ampicillin was active against the P . multocida, S . suis, and S . equi isolates tested (MIC90s, 0.5, 0.06, and 0.06 micrograms/ml, respectively) and was moderately active against S . typhimurium (MIC90s, 2.0 micrograms/ml) . However, this antimicrobial agent was much less active when it was tested against A . pleuropneumoniae, S . cholerae-suis, and E . coli (MIC90s, 16.0, > 32.0, and 32.0 micrograms/ml, respectively) . Against the U.S . isolates of A . pleuropneumoniae and P . multocida, tilmicosin was moderately active (MIC90s, 4.0 and 8.0 micrograms/ml, respectively) . However, this compound was not active against the remaining U.S . isolates (MIC90s, > 64.0 micrograms/ml) . Differences in the MICs from one country to another were not detected with enrofloxacin, ceftiofur, or lincomycin for the strains tested, but variations in the MICs of the remaining antimicrobial agents were observed.

Blood, 1995 Aug 15, 86(4), 1301 - 9
Granulocyte colony-stimulating factor versus placebo in addition to penicillin G in a randomized blinded study of gram-negative pneumonia sepsis: analysis of survival and multisystem organ failure; Smith WS et al.; Sepsis is a common cause of morbidity and mortality . Neutrophils are the major defense against bacterial invasion, and granulocyte colony-stimulating factor (G-CSF) augments both neutrophil number and function . In our study, 160 rabbits were inoculated transtracheally with 0.5 mL of a solution containing 10(4) colony forming units per milliliter of Pasteurella multocida . Twenty-four hours later, chest x-rays and quantitative blood cultures demonstrated pneumonia and bacteremia . Therapy was then begun with penicillin G and either recombinant human G-CSF (rG-CSF; 5 to 8 micrograms/kg subcutaneously) or placebo every day for 5 days . Arterial blood gases and 23 other parameters of organ function were performed before inoculation and serially thereafter . All rabbits underwent histologic examination of organs at the time of septic death or when sacrificed on day 6 . A total of 149 rabbits survived long enough to initiate therapy . A significant increase in leukocytes by day 4 was found in the rG-CSF-treated group . There was a trend towards improved survival in the rG-CSF group (77% v 67%; P = .13, n = 149) . Analysis of pretreatment variables revealed sepsis-induced leukopenia (< or = 2,800/microL) as the only predictor of significantly improved survival with rG-CSF treatment (57% v 39%; P = .04, n = 73) . The majority of the survival benefit occurred within the first 24 hours of treatment . This was before the time that a significant difference in mean white blood cell (WBC) count was observed between the study groups, making intravascular leukocytosis an unlikely explanation for the survival advantage in the rG-CSF group . No significant difference in laboratory variables reflecting organ function was demonstrated between the groups . Histologic grading of inflammation (0, normal, to 6, necrosis) in seven organs revealed that the surviving rabbits had mild but statistically significant increased inflammation in the liver, spleen, and noninoculated lung in the rG-CSF versus placebo groups (liver: 2.6 v 1.5, P < or = .0001; spleen: 3.2 v 2.3, P < or = .0001; and noninoculated lung: 2.9 v 2.5, P = .04) . Administration of rG-CSF, in addition to penicillin G, in immune competent rabbits with gram-negative sepsis complicated by leukopenia significantly improved survival over antibiotics alone . The administration of rG-CSF in early sepsis for a short therapeutic duration was not associated with any clinically evident toxicity . Clinical trials using rG-CSF in septic patients with leukopenia are indicated.

Gene, 1995 Aug 8, 161(1), 39 - 43
Characterization of the Pasteurella multocida skp and firA genes; Delamarche C et al.; A 2.9-kb fragment of the Pasteurella multocida (Pm) genome encoding proteins p25 (25 kDa) and p28 (28 kDa) has previously been cloned and expressed in Escherichia coli (Ec) . In the present paper, the nucleotide (nt) sequence of a 1.8-kb subfragment encoding the two proteins is described . The cloned fragment contains three open reading frames (ORFs) . ORF1 is incomplete . ORF2 is homologous to the skp gene of Ec . ORF3 overlaps ORF2 and is highly homologous to the firA gene of Ec . The skp and firA genes are part of an operon governing the first steps of lipid A synthesis . Comparing the nt sequence with the N-terminal sequences of p25 and p28 revealed that the two proteins are encoded by ORF2 (skp) . The preprotein p28 is converted into p25 by cleavage of a 23-amino-acid leader peptide . Though it serologically cross-reacts with porin H of Pm, p25 is not related to known bacterial porins.

J Vet Pharmacol Ther, 1995 Aug, 18(4), 284 - 9
Antibacterial activity of marbofloxacin . A new fluoroquinolone for veterinary use against canine and feline isolates; Spreng M et al.; Marbofloxacin is a new fluoroquinolone developed exclusively for veterinary use . Minimum inhibitory concentrations of marbofloxacin were assessed for 816 recent isolates associated with canine or feline diseases . Marbofloxacin showed a broad spectrum of activity against gram-negative and gram-positive bacteria . In vitro rates of killing of marbofloxacin and enrofloxacin were compared against strains of Staphylococcus intermedius and Pasteurella multocida, and the results showed no marked difference between the two antibiotics . The duration of bactericidal activity was evaluated ex vivo in the urine of dogs and cats treated with marbofloxacin and lasted from 2 to 5 days after a single administration according to the dosages . Post-antibiotic effect durations were determined with Escherichia coli, Pasteurella multocida, Staphylococcus aureus and Staphylococcus intermedius and were found almost equal to those of enrofloxacin or ciprofloxacin . These results predict a great potential for marbofloxacin in the treatment of a wide range of diseases in dogs and cats.

Zentralbl Veterinarmed B, 1995 Aug, 42(6), 377 - 83
In vitro efficacy of cefquinome (INN) and other anti-infective drugs against bovine bacterial isolates from Belgium, France, Germany, The Netherlands, and the United Kingdom; Bottner A et al.; The in vitro antibacterial activity of cefquinome (INN), an aminothiazolyl-cephalosporin of the fourth generation of cephalosporins, was investigated by determining the minimal inhibitory concentration (MIC, microgram/ml) for 714 bacterial isolates of bovine origin and comparing it with those of amoxicillin, amoxicillin and clavulanic acid, ceftiofur, cephapirin, enrofloxacin, gentamicin, kanamycin and oxytetracycline . Drug resistance was determined by using break-points, which consider the dosage regimen and pharmacokinetics of the veterinary antimicrobials investigated . Cefquinome demonstrated a very high in vitro activity against bacterial isolates of Pasteurella spp., Escherichia coli and Salmonella spp . Overall, the level of resistance against the different anti-infectives tested was lowest for cefquinome . For the remaining substances examined, in vitro activity and the level of resistance showed considerable differences . The chemical and pharmaceutical features of cefquinome are discussed.

Tierarztl Prax, 1995 Aug, 23(4), 398 - 401
{Treatment of orbital abscesses and phlegmon in dogs and cats}; Ruhli MB et al.; A diagnosis of orbital cellulitis or abscess was made in 13 dogs and four cats over the past five years . A foreign body was found in three of these cases . In five cases pasteurella spp . was isolated . In 15 of these cases the abscess was drained surgically . One dog was permanently blind due to inadequate surgical drainage of the abscess . In the remaining cases healing was uneventful . The surgical and medical therapy of orbital abscesses is illustrated by an exemplary case.

Am J Vet Res, 1995 Aug, 56(8), 1055 - 61
Enzyme release by bovine neutrophils; Watson GL et al.; Release of enzymes from cytoplasmic granules has been postulated to have a major role in neutrophil-mediated tissue injury . Secretion or release of primary granules, specific granules, and cytosolic enzymes by bovine neutrophils was examined by quantifying the release of beta-glucuronidase, B12-binding protein, and lactate dehydrogenase, respectively, in response to predetermined amounts of phorbol myristate acetate, calcium ionophore, and opsonized zymosan . These responses were compared with the enzyme release induced by exposure to live or dead, unopsonized or opsonized Pasteurella haemolytica . The greatest release of beta-glucuronidase, B12-binding protein, and lactate dehydrogenase was observed in neutrophils exposed to live organisms partially because of neutrophil lysis . Bovine neutrophils respond markedly to particulate agonists, live or dead, pathogenic or nonpathogenic, by a selective release of specific granules, an effect enhanced by opsonization . Particulate agonists induce minimal primary granule release other than that induced by cell death . Because bovine neutrophils contain quantitatively high numbers of specific granules, the high rate of secretion/release in response to P haemolytica organisms could have a major role in the tissue responses that characterize the lesions of pneumonic pasteurellosis.

Am J Vet Res, 1995 Aug, 56(8), 1045 - 54
Definition of chemiluminescence and superoxide production responses of bovine neutrophils to selected soluble and particulate stimulants, and comparisons with the responses to Pasteurella haemolytica; Watson GL et al.; We defined methods for use of luminol-dependent chemiluminescence (LDCL) and superoxide anion (O2-) production as parameters of the oxidative metabolism of neutrophils isolated from 1.5- to 5-week-old neonatal calves . We determined how variations in blood sample handling, agonist preparation, individual variability, and age of calves influenced the LDCL and O2- responses to certain agonists, and defined concentrations of soluble and particulate agonists that maximally stimulated the oxidative metabolism of bovine neutrophils . Oxidative responses, particularly LDCL, were characterized by marked day-to-day variability, differed greatly within and between calves, were partially age-dependent, and were partially dependent on the individual agonist . Superoxide anion production had substantially less variability . We compared the in vitro oxidative (LDCL and O2-) responses of neutrophils isolated from neonatal calves stimulated by defined concentrations of the agonists--latex, phorbol myristate acetate, calcium ionophore, and opsonized zymosan--with responses to formylated oligopeptides and zymosan-activated serum, and to live, dead, live opsonized, and dead opsonized Pasteurella haemolytica organisms . Opsonization of particulates, pathogenic or nonpathogenic, enhanced the LDCL and O2- responses of stimulated neutrophils although P haemolytica was a less potent stimulant of oxidative functions than were nonbiological agonists . We conclude that the generation of reactive oxygen species by bovine neutrophils in response to P haemolytica is highly dependent on the presence of opsonins and is greatly enhanced in live vs killed bacteria . Furthermore, the in vitro generation of reactive oxygen species, including O2- by stimulated neutrophils, may be of biologic importance if similar events occur in vivo, and could have a major role in the pathogenesis of the acute lung injury associated with pneumonic pasteurellosis.

Am Fam Physician, 1995 Aug, 52(2), 479 - 85, 489-90
Management of cat and dog bites; Lewis KT et al.; An estimated 1 to 2 million Americans are bitten by cats and dogs each year . Most victims are children who are bitten by dogs . Dog and cat bite wounds may appear trivial, but if they are not managed appropriately, they can become infected and may result in functional impairment . Cat bite wounds on the hand have the greatest risk of infection . Pasteurella multocida, isolated in over half of all cat bite wounds and in 20 to 30 percent of dog bite wounds, can cause serious infection with severe complications . Amoxicillin-clavulanate is recommended for the outpatient treatment of dog and cat bite wounds . Tetanus immunization status and the risk of rabies infection should be routinely addressed in bite wound management.

Infect Immun, 1995 Aug, 63(8), 3209 - 12
Hemolytic activity of the Pasteurella haemolytica leukotoxin; Murphy GL et al.; A Pasteurella haemolytica mutant incapable of producing leukotoxin was created by allelic replacement . Concentrated culture supernatants from wild-type P . haemolytica, but not from the mutant, contained the 102-kDa leukotoxin protein and lysed bovine lymphoma cells and sheep erythrocytes . Wild-type P . haemolytica demonstrated the typical beta-hemolytic phenotype on sheep and rabbit blood agar, whereas the mutant did not.

J Clin Pathol, 1995 Aug, 48(8), 775 - 7
Waterhouse-Friderichsen syndrome complicating primary biliary sepsis due to Pasteurella multocida in a patient with cirrhosis; Ip M et al.; Pasteurella multocida is an opportunistic pathogen causing bacteraemia in patients with liver dysfunction . A fulminant case of acute cholecystitis and septicaemia caused by P multocida, complicated by Waterhouse-Friderichsen syndrome without skin haemorrhage, is reported in a previously healthy 64 year old Chinese woman . The patient presented with a six hour history of sudden onset epigastric pain, vomiting, chills, and rigors . A presumptive diagnosis of cholangitis with septicaemic shock was made . Disease progression was rapid and the patient died within eight hours of symptom onset . This case is further proof that skin and mucosal haemorrhages are not an essential feature of Waterhouse-Friderichsen syndrome and this condition should be suspected in all patients presenting with sudden illness and fulminant septicaemia.

Vet Microbiol, 1995 Aug, 45(4), 319 - 29
Partial characterization of the leukotoxin of Pasteurella haemolytica-like bacteria isolated from swine enteritis; DeSilva RT et al.; Pasteurella haemolytica-like (PHL) strains isolated from diarrheic pigs are known to produce a leukotoxin that is lethal to ruminant leukocytes . In the present study, 12 PHL strains were screened for leukotoxin production using a tetrazolium dye-reduction assay . Sterile culture supernatant from strain 6213A, the maximum leukotoxin producer, was used as the crude leukotoxin for characterization studies . The leukotoxin was inactivated by heat at 60 degrees C and by trypsin, protease, and amylase . Toxicity was retained over a pH range of 3.0-11.0 . The leukotoxin was lethal to polymorphoneutrophils (PMNs) of cattle, sheep, goat, and swine . Chromosomal DNA of all 12 PHL strains hybridized with a 3.9 kb Pasteurella haemolytica A1 leukotoxin probe, indicating similarities between the leukotoxin genes of P . haemolytica and PHL strains.

J Reprod Med, 1995 Aug, 40(8), 603 - 5
Pasteurella multocida isolation from a tuboovarian abscess . A case report; Lukban JC et al.; BACKGROUND: Human infection with Pasteurella multocida occurs most commonly following a nonhuman animal bite wound or cat scratch, with local skin and soft tissue infections the most frequent posttraumatic manifestations . Genitourinary tract pathology attributable to this organism occurs infrequently, with only one previous reported case of P multocida infection presenting in the form of a tuboovarian abscess . CASE: A 44-year-old woman with an acute abdomen underwent exploratory laparotomy, revealing a right-sided tuboovarian abscess . Following a total abdominal hysterectomy and bilateral salpingo-oophorectomy with a seven-day postoperative course of intravenous metronidazole, ampicillin and gentamicin, the patient was sent home on a regimen of cephradine, only to return with sepsis three days later . A course of intravenous aqueous penicillin sodium, metronidazole and cefotaxime was administered for the treatment of P multocida sepsis since this organism was identified in the intraoperative pelvic fluid culture 24 hours after the patient's initial discharge . After 13 days of the above regimen, the patient achieved full defervescence and was discharged on hospital day 15 . CONCLUSION: In the setting of a tuboovarian abscess, the clinician should consider P multocida as a potential etiologic agent, especially in a patient with extensive exposure to nonhuman animals . In the treatment of an acute adnexal infection secondary to this organism, one should employ perioperative therapy with the appropriate antibiotics for a duration of at least 14 days.

Biochem Biophys Res Commun, 1995 Jul 26, 212(3), 981 - 7
Fetuin is a substrate for Pasteurella haemolytica O-sialoglycoprotease; Tabatabai LB; A soluble bovine glycoprotein, fetuin, was used as an alternative substrate to identify O-sialoglycoprotease activity in culture supernatant protein fractions of Pasteurella haemolytica . An aliquot of a 24-hour incubation mixture containing fetuin and O-sialoglycoprotease was denatured and examined after gradient sodium dodecyl polyacrylamide gel electrophoresis . The Coomassie-Brilliant-Blue-stained gel was examined for the disappearance of the fetuin band at an apparent molecular mass of 64 KDa . Four major hydrolysis products were identified: an N-terminal fragment of 45 kDa, a 20 kDa fragment at Val50, and two C-terminal fragments at Val273 and His287.

Berl Munch Tierarztl Wochenschr, 1995 Jul, 108(7), 249 - 52
{Proteins from the outer membrane of Pasteurella--1 . Polyacrylamide gel electrophoresis of outer membrane proteins--a possibility for differentiation of Pasteurella haemolytica strains}; Feist H et al.; Using the sodium laurylsarcosinate method, outer membrane proteins (OMPs) of 63 Pasteurella haemolytica strains are isolated and their protein patterns obtained by SDS polyacrylamide gel electrophoresis (SDS-PAGE) are compared . A high degree of similarity became evident both within A and T biotypes in the molecular weight range of 25 to 50 KDa . Strain-to-strain variations are mainly limited to quantitative differences in individual protein bands . It is concluded that within both the A and T biotype groups equal sets of major proteins are present with differing amounts of individual constituents . Biotypes A and T can be clearly distinguished on the basis of marked variations in OMP profiles . OMPs of those AT strains included in comparative studies are exhibiting no general homogeneity, but a relative heterogeneity due to different protein compositions, which are, however, clearly distinct from the patterns of biotypes A and T . It is established unambiguously that isolates belonging to biotypes A or T, as well as AT strains, can be clearly distinguished from each other on the basis of SDS-PAGE analysis of OMP extracts . Thus, polyacrylamide gel electrophoresis of outer membrane proteins can be used for differentiation.

J Wildl Dis, 1995 Jul, 31(3), 358 - 63
Recovery of Pasteurella multocida from experimentally-exposed freshwater snails; Miller SL et al.; We determined how long Pasteurella multocida could survive in experimentally-exposed freshwater snails . Physa virginea were collected from the Sacramento National Wildlife Refuge, Glenn County, California (USA), an enzootic site for avian cholera . Exposure to water containing up to 10(7) P . multocida per ml did not produce observable changes or mortality in snails . A minimum of 84 P . multocida per snail was necessary for detection among the normal snail bacterial flora . When snails were exposed to P . multocida in vials containing 10(7) bacteria per ml, P . multocida was detected for up to 72 hours in snails . When uninoculated snails were placed in aquaria containing 10(6) P . multocida per ml, P . multocida was not detected within the snails; further, P . multocida was detected in the water for only 24 hours at this level . Based on these results, we propose that P . virginea is not an effective reservoir for P . multocida.

Avian Dis, 1995 Jul-Sep, 39(3), 587 - 93
Pasteurella multocida isolated from wild birds of North America: a serotype and DNA fingerprint study of isolates from 1978 to 1993; Wilson MA et al.; Serotype and DNA fingerprint methods were used to study Pasteurella multocida isolated from 320 wild birds of North America . Isolates were collected during 1978-93 . The HhaI profiles of 314 isolates matched the HhaI profile of somatic reference type 1, stain X-73; somatic type 1 antigen was expressed by 310 isolates, and the serotype of four isolates was undetected . Differentiation of the 314 isolates was observed by digestion of DNA with HpaI . None of the HpaII profiles matched the HpaII profile of X-73 (designated HhaI 001/HpaII 001) . Three HpaII profiles were recognized among the somatic type 1 isolates: HpaII 002 (n = 18), HpaII 003 (n = 122), and HpaII 004 (n = 174) . Profile HpaII 002 was found among isolates collected during 1979-83 . Profile HpaII 003 was identified from isolates collected during 1979-89, with the exception of two isolates in 1992 . The HpaII 004 profile was identified from isolates collected during 1983-93 . Of the six remaining isolates, four expressed somatic type 4 and had HhaI profiles identical to the somatic type 4 reference strain P-1662 profile (designated HhaI 004); these isolates were differentiated by digestion of DNA with HpaII . One isolate was identified as serotype F:11, and another was serotype A:3,4 . In the present study, 314 of 316 (99.4%) isolates from wild birds in the Central, Mississippi, and Pacific flyways during 1978-93, were P . multocida somatic type 1.

Avian Dis, 1995 Jul-Sep, 39(3), 489 - 98
Experimental reproduction of endocarditis with Pasteurella gallinarum in mature leghorn chickens; Tjahjowati G et al.; The pathogenicity of Pasteurella gallinarum for mature leghorn chickens was investigated by inoculating thirty 52-week-old chickens intravenously with live P . gallinarum . Each chicken was inoculated once daily for 5 days at one of three different dosage levels with either the type strain ATCC 13361 or a field isolate from a chicken with endocarditis . Chickens were necropsied after death or euthanasia . Valvular endocarditis was present in seven chickens given the field isolate and five chickens given the type strain . Other lesions detected were myocarditis, hepatic and splenic infarcts, nephritis, pneumonia, and encephalitis . At necropsy, P . gallinarum was reisolated from hearts, livers, spleens, lungs, kidneys, and blood . Controls injected with sterile broth had no lesions of endocarditis, nor was P . gallinarum isolated from them . The results confirm the pathogenicity of P . gallinarum for the heart valves of mature chickens.

Avian Dis, 1995 Jul-Sep, 39(3), 451 - 7
The effect of plasmid acquisition on potential virulence attributes of Pasteurella multocida; Lee MD et al.; Several plasmids that were isolated from complement-resistant Pasteurella multocida or Escherichia coli were evaluated for phenotypic markers . Plasmid p2267, isolated from a tetracycline-resistant, complement-resistant fowl cholera field isolate of P . multocida (PM2267), was used to transform a K-12 E . coli (C600); this resulted in increased complement resistance, which was eliminated by curing . Either of two plasmids (p1870 or p70-1, isolated from P . multocida and E . coli, respectively) conferred an increase in complement resistance and invasiveness to turkey epithelial cells when expressed in the Clemson University (CU) vaccine strain of P . multocida . Additionally, the CU strain containing p1870 was more virulent in turkey challenge, and the plasmid appeared amplified in vivo . No detectable differences in major outer-membrane proteins, capsule, or carbohydrate fermentation were found to be associated with the acquisition of these plasmids.

Vet Immunol Immunopathol, 1995 Jul, 47(1-2), 101 - 10
Serum antibody responses of cattle to iron-regulated outer membrane proteins of Pasteurella haemolytica A1; Confer AW et al.; Serum antibody responses to the 70, 77, and 100 kDa iron-regulated outer membrane proteins (IROMPs) of Pasteurella haemolytica A1 were studied in cattle vaccinated with outer membrane protein (OMP) enriched outer membrane fraction, IROMP-enriched outer membrane fraction or live P . haemolytica . Vaccination with an IROMP-enriched outer membrane fraction stimulated antibodies to the 70 kDa IROMP, whereas vaccination with live P . haemolytica stimulated antibodies to the 70 and 77 kDa IROMPs . In a second experiment, sera were used from cattle vaccinated with live or killed P . haemolytica and subsequently challenged . Significant antibody responses to OMP- and IROMP-enriched outer membrane fractions were detected by an enzyme-linked immunosorbent assay (ELISA) for cattle vaccinated with bacterins or live P . haemolytica . Regression analysis indicated significant correlations between high antibody responses to the OMP- or IROMP-enriched fraction and resistance to challenge . Antibody responses to the 70 and 77 kDa IROMPs were significantly greater for the live P . haemolytica vaccinates than for PBS control vaccinates . There was no significant correlation between antibody responses to individual IROMPs and resistance or susceptibility to challenge . These data suggest that antibodies to IROMPs alone are probably not responsible for protective immunity against pneumonic pasteurellosis . Antibodies to IROMPs, however, in conjunction with antibodies to other surface antigens probably enhance immunity to P . haemolytica challenge.

J Infect, 1995 Jul, 31(1), 51 - 3
Pasteurella multocida--an uncommon cause of obstetric and gynaecological sepsis; Riley UB et al.; Pasteurella multocida is a common cause of wound infection following animal-inflicted wounds, but is a rare cause of female genito-urinary sepsis . We present a case of vulval sepsis and a case of intrapartum septicaemia with this bacterium . These two cases indicate that Pasteurella multocida can occasionally colonise the female lower genital tract and this bacterium should be considered in the differential diagnosis of serious infection related to this site.

Can J Vet Res, 1995 Jul, 59(3), 179 - 82
Comparison of protection of experimentally challenged cattle vaccinated once or twice with a Pasteurella haemolytica bacterial extract vaccine; Conlon JA et al.; Three groups of calves (15-18 per group) were injected twice at a 3-week interval with 2 doses of phosphate buffered saline (PBS, CONTROL group), 2 doses of PRESPONSE, a Pasteurella haemolytica A1 bacterial extract vaccine (PRESPONSE-2 group) or 1 dose of PBS followed by a 2nd vaccination with 1 dose of PRESPONSE (PRESPONSE-1 group) . Three weeks after the 2nd vaccination, the calves were challenged intratracheally with P . haemolytica A1 . Calves were evaluated clinically for 3 days prior to challenge and for 5 days after challenge . Six days postchallenge, calves were either euthanized or sent to slaughter and the lungs were evaluated for percent pneumonic tissue . There was a significant effect of single or double application of vaccine on clinical scores (P = 0.0409) . Percent pneumonic tissue at necropsy was significantly affected by vaccine group (P = 0.014) . Calves in the CONTROL group had significantly higher percent pneumonic tissue after arcsine transformation (45.30%) than calves in any group receiving PRESPONSE, regardless of vaccination frequency (25.18% and 25.78%, for calves receiving 2 doses or 1 dose of PRESPONSE, respectively) . Both serum toxin neutralizing and direct agglutinating titers were negatively correlated with percent pneumonic tissue . Most importantly, 1 dose of PRESPONSE was as efficient as 2 doses at eliciting a protective immune response . It is concluded that the presence of P . haemolytica as a natural commensal in the upper respiratory tract of the calf can effectively prime the animal, and allow the animal to respond in an anamnestic nature to only 1 dose of this vaccine.

J Antibiot (Tokyo), 1995 Jul, 48(7), 671 - 5
Synthesis, antimicrobial activity and in vivo fluorine NMR of a hexafluorinated derivative of tilmicosin; Creemer LC et al.; A new fluorinated analog of tilmicosin was synthesized by the reductive amination of desmycosin with 3,5-bis(trifluoromethyl)piperidine . Despite an apparently small change in structure, the fluorinated analog had much less in vitro antimicrobial activity than tilmicosin and it failed to protect 3-day old chicks against a Pasteurella multocida challenge at 64 mg/kg sc . In a preliminary in vivo fluorine NMR experiment in a female Sprague-Dawley rat, a 19F NMR signal was detected in the liver one hour after ip administration of the fluorinated compound . Therefore, although this fluorinated derivative had less antimicrobial activity than tilmicosin, it may nevertheless provide a suitable model of tilmicosin for pharmacokinetic studies using in vivo fluorine NMR.

Bone, 1995 Jul, 17(1), 5 - 9
Cytokine production by calvariae of osteopetrotic mice; Felix R et al.; In the osteopetrotic op/op mouse, the absence of macrophage colony-stimulating factor (M-CSF) prevents the growth of macrophages and osteoclasts and, consequently, bone resorption . In the present study, we investigated whether this deficiency in M-CSF production alters the production of cytokines in op/op bones . Calvariae of phenotypically normal (+/?) and op/op mice were stimulated in vitro with lipopolysaccharide or Pasteurella multocida toxin to produce cytokines . Interleukin-3 (IL-3) and granulocyte-macrophage colony-stimulating factor (GM-CSF) synthesis was the same both in calvaria from osteopetrotic and phenotypically normal animals . However, the production of granulocyte colony-stimulating factor (G-CSF), interleukin-6 (IL-6), and tumor necrosis factor-alpha (TNF alpha) was lower in calvaria from op/op animals than was the case in +/? calvaria . Thus, the lack of biologically active M-CSF causes defects which are not compensated by cells independent of M-CSF.

Am J Vet Res, 1995 Jul, 56(7), 875 - 9
Vaccination of cattle with outer membrane protein-enriched fractions of Pasteurella haemolytica and resistance against experimental challenge exposure; Morton RJ et al.; Administration of an N-lauroylsarcosine-derived outer membrane protein fraction of Pasteurella haemolytica A1 (SCI-1) induced a protective response in calves against intrathoracic challenge exposure with the homologous serovar . Outer membrane proteins from heterologous serovars, A6 and A9, induced partial protection that was associated with their respective similarities to serovar A1 in outer membrane protein profiles derived by use of sodium dodecyl sulfate-polyacrylamide gel electrophoresis . Calves vaccinated with SCI preparations did not have detectable neutralizing antibody to P haemolytica A1 leukotoxin . Antibodies to whole-cell antigens, carbohydrate-protein subunit antigen, and SCI-1 were associated with resistance, which indicates that protein antigens shared among cell surface, carbohydrate-protein subunit, and SCI preparations are immunogenic and enhance resistance to experimental challenge exposure.

Am J Vet Res, 1995 Jul, 56(7), 866 - 9
Colonization of the tonsils and nasopharynx of calves by a rifampicin-resistant Pasteurella haemolytica and its inhibition by vaccination; Frank GH et al.; A rifampicin-resistant Pasteurella haemolytica serotype 1 with 2 added plasmids was used as a colonization-challenge strain in calves to test the resistance to colonization elicited by vaccination . Nine calves were vaccinated with a tissue culture-derived P haemolytica serotype-1 vaccine which, in a prior study, had elicited a serotype-specific inhibition of nasal and tonsillar colonization by the homologous serotype under field conditions . The vaccinates and 9 nonvaccinated control calves were exposed by tonsillar instillation with the challenge strain . The P haemolytica were enumerated in nasal secretion and tonsil wash specimens collected biweekly for 3 weeks . Rifampicin-supplemented agar medium inhibited growth of other bacterial species in the specimens and, thus, increased the sensitivity of detection of the challenge P haemolytica by 100-fold . The challenge strain retained its plasmids during the period of colonization . Inhibition of colonization was evidenced by lower frequency of isolations and fewer isolations of the challenge strain from nasal secretion and tonsil wash specimens of the vaccinates than from those of the nonvaccinates.

Vet Microbiol, 1995 Jul, 45(2-3), 201 - 9
Chaotropic agents cause disaggregation and enhanced activity of Pasteurella haemolytica leukotoxin; Clinkenbeard KD et al.; Pasteurella haemolytica biotype A, serotype 1 grown to late logarithmic growth phase in cell culture medium (RPMI 1640) produced highly aggregated leukotoxin . The multimer mass of the highly aggregated leukotoxin in 0.1 M sodium phosphate buffer pH 7.0 as determined by gel filtration chromatography on Sephacryl S400HR was approximately 8000 kDa . Resuspension of leukotoxin in phosphate buffer containing various chaotropic agents resulted in partial disaggregation of leukotoxin and enhanced leukotoxic activity . 3M guanidine disaggregated leukotoxin to a multimer mass of approximately 800 kDa and enhanced leukotoxic activity 3 to 20-fold . In 6 M urea or 1 M sodium thiocyanate, leukotoxin multimers were observed ranging in mass from 8000 kDa to 400 kDa, and activity enhancement was less than that for leukotoxin in 3 M guanidine . Several detergents were tested for enhancement of leukotoxic activity, but only 1% Tween 20 enhanced leukotoxic activity (4-fold), whereas 1.25% octylglucoside, 10 mM CHAPS, and 5 mM deoxycholate diminished and 1% Triton X-100 abolished leukotoxic activity.

Vet Microbiol, 1995 Jul, 45(2-3), 191 - 200
Colorimetric assay using XTT for assessing virulence of avian Pasteurella multocida strains; Choi KH et al.; A colorimetric assay using sodium 3,3'-{1{(phenylamino)carbonyl}3,4- tetrazolium}-bis(4-methoxy-6-nitro) benzene sulfonic acid hydrate (XTT) was adapted to quantitate bactericidal activity of chicken macrophage HD 11 cell line against five Pasteurella multocida strains and an avirulent transposon insertion mutant . The strains used were virulent P1059, and D92, and four avirulent strains including a streptomycin resistant mutant of P1059 (P1059 SmR), two live vaccine strains namely, the Clemson University (CU) and M9, and a transposon insertion mutant PmTn-294 . Percentage of bacteria killed by chicken macrophage (HD 11) cells was determined by extrapolation from a standard formazan curve derived by incubating XTT with known bacterial cell numbers of each strain . The amount of formazan as measured by absorption at 450 nm was directly related to the number of viable bacterial cells . The percentages of P1059 SmR, CU, M9 and PmTn-294 killed by HD 11 cells were approximately 50%, 61%, 25% and 34%, respectively . By contrast, the virulent P1059 and D92 strains were resistant to killing, and were able to replicate inside the HD 11 cells . Association of virulence with resistance to phagocytic killing by HD 11 cells as assessed by the colorimetric bactericidal assay, was validated with resistance to complement (C')-mediated killing and a turkey mortality test . Strains P1059 and D92 were resistant to C'-mediated killing, whereas strains P1059 SmR, CU, M9 and PmTn-294 strains were susceptible . All turkeys challenged with P1059 or D92 were dead within 18 hrs . Mortality did not occur in turkeys challenged with strains of P1059 SmR, M9 and PmTn-294 . The mortality among CU challenged turkeys ranged from 0 to 40% . The results suggest that the colorimetric bactericidal assay using XTT can be used to quantitate chicken macrophage phagocytic killing of P . multocida strains, and may be a valuable assay to differentiate virulent from avirulent strains of avian P . multocida.

Lab Anim, 1995 Jul, 29(3), 314 - 9
Reclassification of 30 Pasteurellaceae strains isolated from rodents; Boot R et al.; Thirty Pasteurellaceae strains isolated from gerbil, guineapig, hamster, mouse, muskrat and rat were reinvestigated and reclassified after comparison with reference strains . Strains originally described as Pasteurella pneumotropica were reclassified as {Pasteurella} pneumotropica Heyl biotype (7), {P.} pneumotropica Jawetz biotype (1), Pasteurella dagmatis (1) or Taxon 22 (2) . Strains previously reported as Actinobacillus sp . were reclassified as {P.} pneumotropica biotype Jawetz (3), P . dagmatis (3) or Taxon 6 (7) . Strains earlier described as Pasteurella gallinarum were renamed as SP group pasteurella (4) or Taxon 25 (2) . Some of these reclassified Pasteurellaceae have not been reported previously in rodents . The present findings underline the importance of extended characterization of isolates and comparison with references strains to avoid misclassification within the family Pasteurellaceae Pohl 1981.

Pediatrics, 1995 Jun, 95(6), 944 - 8
First case of human infection caused by Pasteurella gallinarum causing infective endocarditis in an adolescent 10 years after surgical correction for truncus arteriosus; al Fadel Saleh M et al.; OBJECTIVE . To report the first case of human infection (infective endocarditis {IE}) caused by Pasteurella gallinarum and to review the literature regarding IE caused by the genus Pasteurella . SETTING . University hospital based . PATIENT . An adolescent boy who underwent successful correction for truncus arteriosus 10 years before the present illness . RESULTS . Persistent fever, pallor, and a palpable spleen suggested IE clinically . Echocardiography documented vegetation in the conduit that was used for surgical correction . Blood cultures grew P . gallinarum and confirmed its role as the causative organism for IE in the patient . CONCLUSION . This case illustrates that IE may develop in a child with congenital heart disease several years after surgical intervention using material that is foreign to the body (conduit), and that such a complication may involve unusual pathogens . These observations emphasize the need for careful long-term follow-up of children with congenital heart disease even after successful surgical correction.

J Anim Sci, 1995 Jun, 73(6), 1658 - 65
Effects of atrophic rhinitis induced by Pasteurella multocida toxin on heat production and activity of pigs kept under different climatic conditions; van Diemen PM et al.; Effects of moderate, artificially induced atrophic rhinitis symptoms on level and changes in heat production and activity were determined in pigs kept under different climatic conditions . Eight groups of 30 pigs each, housed in one of two climatically controlled respiration chambers, were exposed to a 2 x 2 factorial arrangement of treatments: challenged with 0 or 13 micrograms of Pasteurella multocida toxin (Pm-T)/mL, and two climatic environments (good: 25 degrees C, or adverse: 15 degrees C with draught periods) . The Pm-T challenge reduced (P < .05) day averages of total (HP) and activity-related heat production (Har) . The response to Pm-T treatment was similar in both climatic environments . Differences in the heat production and activity caused by the climatic treatment declined (P < .001) with time and acclimation to the environment . Analyses of HP, Har, and activity-free heat production in 12 2-h periods showed a biphasic activity rhythm . Both treatments affected (P < .05) level of HP and Har in several of the 2-h periods, but the biphasic rhythm was not altered . Day averages of Har showed a disposition to be differently affected (P < .068) by Pm-T challenge in the climatic treatments dependent on duration of exposure . This interaction effect (P < .001) seemed to originate from the periods between 1500 and 2100 . Therefore, it might be wise to distinguish between overall effects (day means) on total, activity-related, and activity-free heat production and effects within a day (e.g., 2-h means) . Treatment with Pm-T seemed to suppress the general well-being of pigs, reducing pigs' activity and food intake.(ABSTRACT TRUNCATED AT 250 WORDS)

Infect Immun, 1995 May, 63(5), 1703 - 9
Extracellular neuraminidase production by a Pasteurella multocida A:3 strain associated with bovine pneumonia; White DJ et al.; The properties of an extracellular neuraminidase produced by a Pasteurella multocida A:3 strain that was isolated in a case of bovine pneumonia were examined during growth in a defined medium . This enzyme (isolated from concentrated culture supernatants of P . multocida A:3) was active against N-acetylneuramin lactose, human alpha-1-acid glycoprotein, fetuin, colominic acid, and bovine submaxillary mucin . Enzyme elaboration was correlated with the growth of the organism in a defined medium, with maximum quantities produced in the stationary phase . The enzyme was purified by a combination of ammonium sulfate fractionation, ion exchange on DEAE-Sephacel, and gel filtration on Sephadex G-200 . The purified neuraminidase possessed a specific activity of 9.36 mumol of sialic acid released per min per mg of protein against fetuin . The enzyme possessed a pH optimum of 6.0 and a Km of 0.03 mg/ml . The P . multocida A:3 neuraminidase had a molecular weight of approximately 500,000 as estimated by gel filtration . The enzyme was stable at 4 and 37 degrees C for 3 h . Approximately 75% of the neuraminidase activity was lost within 30 min at 50 degrees C . Greater than 90% of the enzyme activity was destroyed within 10 min at temperatures of > or = 65 degrees C . The P . multocida neuraminidase does not appear to be serologically related to the Pasteurella haemolytica A1 neuraminidase since antiserum prepared against the purified P . haemolytica enzyme did not neutralize the P . multocida enzyme.

J Interferon Cytokine Res, 1995 May, 15(5), 431 - 9
Effect of recombinant bovine interleukin-1 beta on viral/bacterial pneumonia in cattle; Baca-Estrada ME et al.; Bovine herpesvirus-1 (BHV-1) is an important pathogen of respiratory infections in cattle . Its continuing importance lies in its ability to predispose infected hosts to bacterial infections (e.g., Pasteurella haemolytica) . In this study we determined whether the immunoregulatory effects induced by recombinant bovine interleukin-1 (rbIL-1) could stimulate appropriate host defense mechanisms to influence the course of BHV-1 and P . haemolytica infection in cattle . We first evaluated the effect of multiple doses (5 doses of 300 ng/kg) of rbIL-1 in normal cattle . An increase in polymorphonuclear (PMN) cells, as well as monocytes, in peripheral blood was observed during the course of IL-1 administration . In addition, the phagocytic activity of monocytes was increased . Although the phagocytic and oxidative burst activities in PMN decreased during the course of rbIL-1 treatment, no changes were observed in the bactericidal capacity of these cells . Lymphocyte numbers in peripheral blood remained unchanged; however, the functional activity of these cells, as measured by IFN-gamma production upon in vitro stimulation, was decreased . In the bovine respiratory disease model, multiple administration of IL-1 did not influence significantly the progression of BHV-1/P . haemolytica infection in cattle . Thus, our results demonstrated that IL-1, although not therapeutically effective, could be administered safely as an adjuvant, even during the course of BHV-1/P . haemolytica infection.

Br Vet J, 1995 May-Jun, 151(3), 233 - 62
Equine pleuropneumonia; Raidal SL; Pleuropneumonia is a clinically important equine disease, predisposed by a number of identifiable factors . Successful management is largely dependent on early identification and prompt initiation of appropriate treatment strategies . Rapid resolution of the disease process is associated with appropriate treatment commenced within 48 h of the causative insult . Lower airway contamination by oropharyngeal organisms and subsequent extension into the pulmonary parenchyma results in respiratory dysfunction and systemic toxaemia . Acute disease is associated with the isolation of facultatively anaerobic organisms, especially beta-haemolytic Streptococcus spp . and Pasteurellaceae . Delayed or inappropriate treatment is likely to result in chronic disease characterized by the involvement of anaerobic bacteria and a poor response to therapy . The primary mode of treatment for anaerobic infection of the human thorax is surgical drainage and resection of necrotic tissue but whilst such techniques have been described for the management of equine pleuropneumonia, the size of the equine thoracic cavity hinders accurate diagnostic evaluation and successful completion of such intervention . The chronic nature and cost of ongoing treatment and limitations on choice of antimicrobial agents warrant a poor prognosis for survival and a poorer prognosis for return to athletic endeavour.

Antimicrob Agents Chemother, 1995 May, 39(5), 1097 - 100
Comparative in vitro activities of azithromycin, Bay y 3118, levofloxacin, sparfloxacin, and 11 other oral antimicrobial agents against 194 aerobic and anaerobic bite wound isolates; Goldstein EJ et al.; The activities of sparfloxacin, levofloxacin, Bay y 3118, azithromycin, cefprozil, loracarbef, and nine other oral antimicrobial agents against 194 aerobic and anaerobic clinical bite wound isolates were determined by the agar dilution method . Sparfloxacin, levofloxacin, and Bay y 3118 were active against all aerobic isolates (MICs at which 90% of the isolates are inhibited {MIC90}, < or = 1.0 microgram/ml for sparfloxacin and levofloxacin and 0.1 microgram/ml for Bay y 3118) and many anaerobic isolates, with the exception of the fusobacteria . Azithromycin was more active than erythromycin by 1 to 2 dilutions against many aerobes, including Pasteurella multocida and Eikenella corrodens, and by 2 to 4 dilutions against anaerobic isolates . Cefprozil was more active (MIC90, < or = 1 microgram/ml) than loracarbef (MIC90, < or = 4 micrograms/ml) against aerobic gram-positive isolates, but both had poor activity (MIC90, > or = 16 micrograms/ml) against peptostreptococci . Both cefprozil and loracarbef had MIC90s of < or = 0.5 micrograms/ml against P . multocida.

Vet Pathol, 1995 May, 32(3), 274 - 9
Effects of the Pasteurella multocida toxin on osteoblastic cells in vitro; Sterner-Kock A et al.; Pasteurella multocida toxin induces localized osteolysis in the turbinate bones of swine . Osteolysis appears to be due to an increased level of osteoclastic bone resorption, although osteoblast activity may also be impaired . We studied the effects of purified toxin on the osteoblastic phenotype of the ROS 17/2.8 rat osteoblastic osteosarcoma cell line . Treatment of both embryonic bovine lung cells and a nonosteoblastic rat osteosarcoma cell line (ROS 25/1) with nanomolar doses of toxin produced marked cytotoxic actions . In the osteoblastic ROS 17/2.8 cells, this level of toxin reduced expression of an osteoblastic marker (alkaline phosphatase), was associated with matrix mineralization, but had no cytopathologic action . The osteoblastic cell population may be resistant to a direct cytotoxic effect but is nevertheless a target for toxin action.

J Comp Pathol, 1995 May, 112(4), 381 - 9
Predisposition of specific pathogen-free lambs to Pasteurella haemolytica pneumonia by Bordetella parapertussis infection; Porter JF et al.; Three groups of specific pathogen-free (SPF) lambs were inoculated intratracheally with an ovine isolate of Bordetella parapertussis (5.5 x 10(9) colony-forming units) or with B . parapertussis followed 2 or 5 days later with Pasteurella haemolytica serotype A2 (120-180 million colony-forming units) . When P . haemolytica A2 was administered 2 days after infection with B . parapertussis all lambs became febrile for at least 72 h . At necropsy their lungs were discoloured, congested and showed large areas of collapse and consolidation which, in one case, covered the entire lung . Histopathological examination confirmed that the combined infection produced a severe acute bronchopneumonia in four of seven lambs . B . parapertussis and P . haemolytica were recovered from all of the lambs in this group . Seven lambs challenged with P . haemolytica 5 days after B . parapertussis and six lambs infected with B . parapertussis alone showed no clinical signs of disease other than mild pyrexia and only mild histopathological changes . B . parapertussis, but not P . haemolytica, was recovered from these lambs . The findings indicated that B . parapertussis predisposed the SPF lambs to P . haemolytica pneumonia . This effect appeared to be dependent upon the time interval between the administration of the two agents.

FEMS Microbiol Lett, 1995 Apr 15, 128(1), 75 - 80
Genetic transformation of Vibrio anguillarum and Pasteurella piscicida by electroporation; Cutrin JM et al.; Vibrio anguillarum and Pasteurella piscicida are Gram-negative bacteria which are pathogenic for marine fish and we report here the first successful transformation of these two bacteria by electroporation . The optimal conditions for electroporation included a field strength of 12.5 kV cm-1 and a time constant of 5 ms using 0.2-cm cuvettes . With these parameters, three plasmids (pSU2718, pCML, pEV3) with molecular sizes of 2.6, 5 and 13.7 kb, respectively were successfully transformed into both pathogens . V . anguillarum isolates belonging to serotypes O1 and O2 were transformed with greatest efficiency, 2.5 x 10(3) transformants per micrograms DNA, being achieved in the serotype O2 strains using plasmid pCML . Strains of serotype O3 were not transformed . In the case of P . piscicida the maximum efficiency achieved was 9.8 x 10(2) transformants per micrograms pCML plasmid DNA . This optimized system will allow development of procedures for the genetic manipulation of these pathogens.

Biochemistry, 1995 Apr 4, 34(13), 4193 - 201
Secretion and circular dichroism analysis of the C-terminal signal peptides of HlyA and LktA; Zhang F et al.; The secretion of the 107 kDa hemolysin A (HlyA) from Escherichia coli is mediated by membrane proteins hemolysin B (HlyB) and hemolysin D (HlyD) . The signal for transport has been mapped to the C-terminal 60 amino acids of the HylA molecule . We have shown previously that the C-terminal 70 amino acids of leukotoxin (LktA) from Pasteurella hemolytica can substitute functionally for the HlyA signal sequence . This 70 amino acid peptide contains little primary sequence similarity to the HlyA signal sequence, and we have hypothesized that these signal sequences assume a similar higher-order structure which is recognized by the HlyB/D transporter . In the present study, we have expressed and purified small peptides containing the C-terminal 61 amino acids of HlyA and the C-terminal 70 amino acids of LktA . We show that these signal peptides are sufficient for secretion from E . coli in a HlyB/D dependent manner . Circular dichroism analyses show that both molecules exhibit common biophysical properties . In aqueous solution, they appear to be mainly unstructured, but in a membrane mimetic environment they assume a helical secondary structure . The conformational change observed for both peptides going from an aqueous to a membrane mimetic environment may be an important feature of these signal sequences necessary for their recognition and transport.

Rev Latinoam Microbiol, 1995 Apr-Jun, 37(2), 121 - 6
Serotypes of Pasteurella multocida and Pasteurella haemolytica isolated from pneumonic lesions in cattle and sheep from México; Blanco-Viera FJ et al.; A total of 13,000 pairs of lungs were examined at Mexico's City abbatoir, where 8,000 corresponded to sheep and 5,000 to cattle . From those, 224 pneumonic lesions were observed, obtaining 97 positive isolates, which yielded 112 strains of Pasteurella sp . Forty isolates were identified as P . haemolytica and 72 as P . multocida . One-hundred percent of P . haemolytica belonged to biotype A . Serotypes were determined by indirect haemagglutination . P . multocida isolates were classified according to the acriflavine and hyaluronidase techniques, 61% belonged to type A, 25% to type D and 14% were untypified . Somatic serotypes were determined by gel immunodiffusion; serotype 3 was more frequent, in sheep 72% and in cattle 77%.

Infect Immun, 1995 Apr, 63(4), 1340 - 8
Pasteurella haemolytica serotype 2 contains the gene for a noncapsular serotype 1-specific antigen; Gonzalez CT et al.; An ssa1-homologous genomic fragment cloned from Pasteurella haemolytica serotype 2 (ST2) enabled transformation of Escherichia coli DH5 alpha to a serotype 1 (ST1) phenotype through expression of the ST1-specific antigen (Ssa1) . The Ssa1 protein expressed by ssa1-transformed E . coli was susceptible to heat and protease treatment and was distinct from P . haemolytica ST1-specific capsular polysaccharide . Electrophoretic analysis of in vitro-translated proteins, as well as the predicted amino acid sequence, demonstrated that Ssa1 proteins encoded from either ST1- or ST2-derived ssa1 genes were essentially identical . A comparison of the nucleotide sequences of ssa1 genes derived from P . haemolytica ST1 and ST2 revealed greater than 99% homology . Amino acid sequence homology of the predicted products of ST1- and ST2-derived ssa1 genes was greater than 98% . Northern (RNA) blot studies revealed that the presence of an increased level of ssa1 transcript in P . haemolytica ST1 grown as surface-adherent cultures on solid medium was correlated with a serologically detectable Ssa1 protein . Expression of the ssa1 transcript in ST1 was similarly upregulated by a high iron concentration in the growth medium.

Clin Infect Dis, 1995 Apr, 20(4), 1055 - 7
Pasteurella multocida tonsillitis: case report and review; Ramdeen GD et al.; Pasteurella multocida is frequently part of the normal flora of the nasopharynx and digestive tract of several wild and domestic animals . Although P . multocida can produce a variety of upper and lower respiratory tract infections, only four previous cases of tonsillitis caused by this organism have been reported . We present a case of pasteurella tonsillitis in a 30-year-old female who was exposed through her cat, which manifested upper respiratory symptoms.

J Clin Microbiol, 1995 Apr, 33(4), 952 - 7
Identification and characterization of outer membrane proteins of Pasteurella multocida serotype D by using monoclonal antibodies; Marandi MV et al.; Monoclonal antibodies (MAbs) against Pasteurella multocida serotype D were obtained by fusion of spleen cells from BALB/c mice immunized with outer membrane proteins (OMPs) with SP2/0-Ag 14 murine myeloma cells . Desirable MAbs were selected by enzyme-linked immunosorbent assay (ELISA) with OMP as the antigen . MAbs MT1 and MT2 identified two different proteins (H {heavy} and W {weak}), each with a molecular mass of 32 kDa, in Western blots (immunoblots) . Treatment of the OMPs with proteolytic enzymes and sodium periodate indicated that the binding sites of MAbs MT1 and MT2 are of protein and glycoprotein natures, respectively . The epitopes reactive with MAbs were surface exposed, as visualized by immunoelectron microscopy . Among field isolates of P . multocida serotype D, two distinct OMP patterns were recognized by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and these patterns were designated types I and II . In both the ELISA and the Western blot, MAb MT1 recognized only type I isolates, whereas MAb MT2 recognized both type I and II isolates . Neither MAb MT1 nor MAb MT2 reacted with either reference strains of capsular serotypes A, B, E, and F or field isolates of capsular serotype A of P . multocida . This is the first report of MAbs identifying the serotype D-specific OMP of P . multocida.

Can J Vet Res, 1995 Apr, 59(2), 154 - 6
Pasteurella multocida toxin induces IL-6, but not IL-1 alpha or TNF alpha in fibroblasts; Rosendal S et al.; Pasteurella multocida toxin (PMT) is the major virulence factor in Progressive Atrophic Rhinitis of swine . Other workers' previous findings that PMT was mitogenic for 3T3 fibroblasts, were confirmed in the present study . In addition, PMT stimulated 3T3 cells to release IL-6, but IL-1 alpha or TNF alpha were not detected in fibroblast supernatants sampled 24, 48, or 72 h after stimulation . In view of the role of IL-6 in osteoclastic bone resorption, these findings provide a new working hypothesis for investigations into the molecular pathogenesis of this important disease.

Can J Vet Res, 1995 Apr, 59(2), 110 - 7
Dissociation of cytolysis and monokine release by bovine mononuclear phagocytes incubated with Pasteurella haemolytica partially purified leukotoxin and lipopolysaccharide; Stevens P et al.; The bovine respiratory pathogen Pasteurella haemolytica secretes an exotoxin that is specific for ruminant leukocytes (leukotoxin) . Previous studies have shown that subcytolytic concentrations of the leukotoxin stimulate bovine neutrophils to undergo a respiratory burst and degranulate . Relatively little is known about the stimulatory effects of the leukotoxin on bovine mononuclear phagocytes . In this study, we compared the relative cytolytic effects of partially purified leukotoxin on bovine peripheral blood monocytes and alveolar macrophages . We found monocytes to be approximately 8- to 10-fold more sensitive than alveolar macrophages to the cytolytic effect of leukotoxin . In addition, incubation of monocytes and alveolar macrophages with sublethal doses of leukotoxin stimulated release of IL-1 and TNF activities in a dose-dependent manner . Addition of an antileukotoxin MAb neutralized the cytolytic effects of leukotoxin, but potentiated TNF release . Heat inactivation also blocked the cytolytic activity of LKT, but only slightly reduced its ability to stimulate TNF release . Although the leukotoxin preparations were estimated to have only small amounts of lipopolysaccharide (LPS) contamination, as determined by a standard Limulus amebocyte lysate coagulation assay, a chromogenic Limulus assay indicated much greater amounts of LPS were present . Adding equivalent doses of P . haemolytica LPS largely duplicated the monokine release stimulated by leukotoxin . These results suggest that the stimulatory effects of the P . haemolytica leukotoxin on bovine mononuclear phagocytes may principally involve LPS, perhaps complexed with leukotoxin.

J Vet Pharmacol Ther, 1995 Apr, 18(2), 94 - 100
Clinical pharmacology of cefixime in unweaned calves; Ziv G et al.; Cefixime is a unique third-generation oral cephalosporin . Its in vitro activity and pharmacokinetic properties have been studied to assess its potential for use in the therapy of newborn calf infections due to gram-negative bacteria . The minimum inhibitory concentrations of cefixime for 90% (MIC90) of field isolates of Escherichia coli, Salmonella and Pasteurella were 0.10-0.40 micrograms/mL . The serum disposition kinetics of cefixime following intravenous and oral administration was evaluated . The elimination half-life of cefixime after intravenous and oral administration was 3.5-4.0 h, the steady-state volume of distribution was 0.34 L/kg and approximately 90% of the drug was bound to serum proteins . Oral absorption was comparatively slow and bioavailability values for single 5 mg/kg doses were 20.2% after the administration of 200 mg of cefixime in capsules, 28.3% after dosing an aqueous solution of cefixime and 35.7% after fasted calves received the solution of cefixime . Mean serum drug concentrations 12 h after the cefixime solution was administered orally (5 mg/kg) were 1.05 micrograms/mL for the milk-fed calves and 1.76 micrograms/mL for the fasted calves . Computations showed that mean free drug concentrations equal to the MIC50 of the drug for gram-negative pathogens associated with newborn calf infections can be maintained in tissues by multiple treatments at 5 mg/kg every 12 h or 10 mg/kg every 24 h.

Lab Anim, 1995 Apr, 29(2), 192 - 9
Inefficacy of enrofloxacin in the elimination of Pasteurella multocida in rabbits; Mahler M et al.; The administration of enrofloxacin (5 mg/kg subcutaneously every 12 h for 10 days) failed to eliminate Pasteurella multocida from all naturally and experimentally infected rabbits . Although the enrofloxacin concentrations in serum and in turbinate bones were greater than the determined minimal inhibitory concentrations, P . multocida could be detected in nasal cavities, turbinates, trachea, middle ear and outer ear of experimentally infected rabbits after treatment . It is to be supposed that P . multocida colonizes organs or tissues in which an effective enrofloxacin concentration cannot be achieved . Such sites could be the paranasal sinuses, the auditory tube and the middle ear . This finding underlines the indispensibility of in vivo testing of antibiotic effectiveness.

Berl Munch Tierarztl Wochenschr, 1995 Apr, 108(4), 127 - 32
{The resistance behavior of significant veterinary bacterial agents in the year 1992 (ranking in sequence)}; Trolldenier H; On the basis of results of sensitivity tests carried out in 1992 by means of a standardized agar diffusion test according to DIN 58,940 by 28 laboratories performing routine diagnosis, frequency of resistance was evaluated in the form of rank orders . For "calculated" chemotherapy, the choice of the substance to be applied is determined by the sensitivity of the presumptive pathogen, if laboratory results relating to the agent are (still) lacking . Evaluation of the pathogens tested (clostridia, E . coli, pasteurellas, salmonellas, staphylococci, streptococci) has made it evident that, due to a wide distribution of resistance factors against benzylpenicillins, tetracyclines, chloramphenicol, streptomycin, macrolides and sulfonamides, the range of antibiotics for the (uncontrolled) first application has become even narrower . A testing of the pathogen in the antibiogram remains an urgent necessity.

Microb Pathog, 1995 Apr, 18(4), 237 - 52
Purified Pasteurella haemolytica leukotoxin induces expression of inflammatory cytokines from bovine alveolar macrophages; Yoo HS et al.; We obtained biologically active purified leukotoxin (Lkt) from Pasteurella haemolytica serotypel, strain 12296 using preparative sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) . Three species of Lkt of molecular masses 95, 100, and 104 kDa were obtained . Purity of all three species of Lkt was confirmed by analytical SDS-PAGE and Western blot (immunoblot) analysis . Results from the chromogenic Limulus amebocyte lysate assay and silver staining of SDS-PAGE patterns indicated that the preparations were free of contaminating lipopolysaccharide . We then studied the kinetics of TNF alpha and IL-1 beta mRNA expression in bovine alveolar macrophages stimulated with the purified 104 kDa Lkt . Subcytolytic concentrations of Lkt induced TNF alpha and IL-1 beta gene expression and peak induction was observed at a concentration of 1 leukotoxin unit/ml . Both TNF alpha and IL-1 beta mRNA expression were detectable at 1 h after stimulation with 1 leukotoxin unit/ml . The expression peaked at 2 h, steadily declining up to 6 h, and was undetectable by 10 h . Secreted TNF alpha measured by bioassay peaked at 4-6 h and accumulated at a lesser concentration after 6 h . By contrast, secreted IL-1 peaked at 6 h and decreased significantly by 10 h . The ability of purified Lkt to induce TNF alpha and IL-1 beta gene expression and secretion of bioactive proteins was suppressed by Ca2+ chelating agents, 5 mM EDTA and 5 mM EGTA, but not polymyxin B . Heat-inactivation of the purified Lkt that had lost its cytocidal property completely abrogated induction of TNF alpha and IL-1 beta gene expression and secretion in bovine AMs.(ABSTRACT TRUNCATED AT 250 WORDS)

Presse Med, 1995 Mar 18, 24(11), 519 - 22
{Bactericidal activity of ciprofloxacin and sparfloxacin . Comparison with others active antibiotics against Pasteurella multocida}; Gastine C et al.; Bactericidal activities of ciprofloxacin and sparfloxacin were compared to other antibiotics active against human isolates of Pasteurella multocida . Three human isolates of Pasteurella multocida were used for killing-curve studies with ciprofloxacin and sparfloxacin comparatively to others antibiotics . At 2x the MIC, ciprofloxacin and sparfloxacin exhibited a killing of more than 99.9% of the initial viable cells that was achieved within 6 h of incubation . These activities were faster than those of amoxycillin and cefpodoxime . No regrowth was observed after 24 h of incubation . Doxycycline and clarithromycin used at MICx2 had no bactericidal activities . It was concluded that fluoroquinolones, namely ciprofloxacin and sparfloxacin, can be considered having good bactericidal activity against P . multocida.

Infect Immun, 1995 Mar, 63(3), 1033 - 9
Antigenic and virulence properties of Pasteurella haemolytica leukotoxin mutants; Petras SF et al.; Antigenic properties of two mutants of Pasteurella haemolytica, strains 59B0071 and 59B0072, that do not produce detectable leukotoxin were investigated . Western blot (immunoblot) analysis with a number of polyclonal sera from animals recovering from pasteurellosis revealed that both mutants secreted a variety of antigens that were also present in cultures of several wild-type strains . These antigens ranged from about 100 to 15 kDa . Mutant strain 59B0071 was found to be totally deficient in leukotoxin, as judged not only by Western blotting but also by cytotoxicity assays with bovine lymphoma (BL-3) cells or bovine polymorphonuclear cells as targets . The mutant strain 59B0071 had normal levels of a secreted sialylglycoprotease, however . When strains were tested for virulence in goat and cattle challenge experiments, a reduction in mortality and lung lesions was observed with the mutant 59B0071 in comparison with results obtained with wild-type strains . These results are consistent with an important role for leukotoxin in P . haemolytica virulence and suggest that leukotoxin-negative mutants may be useful tools in the investigation of other virulence properties involved in P . haemolytica infections.

Infect Immun, 1995 Mar, 63(3), 1027 - 32
Isolation of Pasteurella haemolytica leukotoxin mutants; Chidambaram M et al.; Two mutants of Pasteurella haemolytica A1 that do not produce leukotoxin were isolated . Following mutagenesis, colonies were screened with antiserum by a filter assay for absence of the secreted leukotoxin . The two mutants both appeared to produce normal amounts of other antigens, as judged by reactivity with polyclonal serum from an animal with pasteurellosis, and were not altered in beta-hemolytic activity as seen on blood agar plates . There was no evidence of either cell-associated or secreted leukotoxin protein when Western blots (immunoblots) were carried out with the polyclonal serum or with a monoclonal antibody directed against the leukotoxin . Southern blots revealed that both mutants show the wild-type restriction pattern at the leukotoxin locus, although the strain with the lktA2 mutation showed differences in other regions of the chromosome on analysis by pulsed-field gel electrophoresis . The strain with the lktA2 mutation grew more slowly than did the wild-type strain, while the strain with the lktA1 mutation was indistinguishable from the wild-type strain in its growth properties . The strain with the lktA1 mutation should be valuable in determining the role of the leukotoxin in virulence as well as in identifying other virulence factors of P . haemolytica.

Arch Microbiol, 1995 Mar, 163(3), 211 - 6
Fatty acid profiles of "Pasteurella" piscicida: comparison with other fish pathogenic gram-negative bacteria; Romalde JL et al.; The fatty acid methyl ester (FAME) profiles of "Pasteurella" piscicida were determined by gas chromatography and subjected to numerical analysis in comparison with those obtained for Vibrio anguillarum, Aeromonas salmonicida and Pasteurella species of clinical origin . The bacterial species studied shared important characteristics with respect to their FAME content: in all of them the saturated and unsaturated fatty acids of 16 carbon atoms were the predominant fatty acids . However, distinguishing features could be detected for each pathogen . Using either single linkage or complete linkage algorithms, strains were divided into four phena that corresponded to the different species, but showed a high degree of correlation among them . Although single linkage discriminated strains better within each phenum, complete linkage was more useful to establish the relationships among clusters . The results obtained support the idea that "Pasteurella" piscicida is related to members of the genera Vibrio and Aeromonas and indicate the need for exhaustive genetic studies to clarify the taxonomic position of this fish pathogen.

Vet Pathol, 1995 Mar, 32(2), 173 - 83
Pasteurella haemolytica lipopolysaccharide-induced cytotoxicity in bovine pulmonary artery endothelial monolayers: inhibition by indomethacin; Paulsen DB et al.; Exposure of bovine pulmonary artery endothelial cells to Pasteurella haemolytica lipopolysaccharide caused severe morphologic changes . Initially, there was dilatation of the rough endoplasmic reticulum and mitochondrial swelling followed by cell retraction, membrane bleb formation, and cell detachment . The affected endothelial cells had severe membrane damage resulting in the leakage of lactate dehydrogenase . Indomethacin in concentrations of 0.5 mM or greater caused marked decreases in the lipopolysaccharide-induced leakage of lactate dehydrogenase . Indomethacin at 5 mM also caused a marked reduction of the lipopolysaccharide-induced morphologic changes resulting in apparent maintenance of the monolayer integrity for 8 hours versus 1 hour in the lipopolysaccharide-treated control . A marked decrease in the cell and nuclear membrane effects resulted, but the rough endoplasmic reticulum dilatation and mitochondrial changes proceeded . These results indicate that indomethacin does not prevent lipopolysaccharide binding but interferes with later events in lipopolysaccharide-induced cytotoxicity in the bovine pulmonary endothelial cell . The concentration of indomethacin required to produce this inhibition suggests that the primary mechanism is not cyclooxygenase inhibition.

Vet Pathol, 1995 Mar, 32(2), 140 - 6
Lectin histochemistry of normal and herpesvirus-infected bovine nasal mucosa; Mosier DA et al.; Proliferation of Pasteurella haemolytica serotype 1 in the nasal cavity following stress or viral infection is an important event in the pathogenesis of bovine pneumonic pasteurellosis . Enhanced adhesion of P . haemolytica to nasal mucosa could be one factor that predisposes animals to this proliferation . Nasal mucosa from normal and bovine herpesvirus-1 (BHV1)-infected cattle were examined histochemically for their glycoconjugate composition . Twenty lectins were screened, six of which were chosen for subsequent study . Three of these were specific for N-acetylgalactosamine (NAGal) (Dolichos biflorus, Glycine max, and Vicia villosa), and one each was specific for N-acetylgalactosamine/galactose (Griffonia simplicifolia-I), mannose/glucose (Canavalia ensiformis), and N-acetylglucosamine (Triticum vulgaris) . For the surface mucosa and submucosal glands, there was greater reactivity in samples from BHV1-infected than from normal cattle for all six lectins . Reactivity was most prominent for the NAGal-specific lectins . Neuraminidase treatment of samples from normal and BHV1-infected cattle tended to result in greater lectin reactivity . Lectin reactivity was generally more intense in focally inflamed areas, but diffuse reactivity was not substantially affected by inflammation . BHV1-induced alteration of nasal mucosal glycoconjugates could enhance adhesion and colonization of P . haemolytica to nasal surfaces and may be one factor responsible for the increased number of P . haemolytica serotype 1 in the nasal cavity following viral infection.

Res Vet Sci, 1995 Mar, 58(2), 163 - 8
A native plasmid of Pasteurella haemolytica serotype A1: DNA sequence analysis and investigation of its potential as a vector; Wood AR et al.; The complete nucleotide sequence of a 4.3 kilobase pair plasmid, pAB2, isolated from a bovine strain of Pasteurella haemolytica serotype A1, was determined . It encodes a Rob-1 type beta-lactamase and a region with homology to the mobilisation (mob) region of the Escherichia coli plasmid, ColE1 . An insertion mutant of pAB2 (pTC2/81) carrying a copy of Tn5 was transferred to E coli K12 by conjugation . Subsequently pTC2/81 could be transferred by transformation to E coli HB101, but not to P haemolytica serotypes A1 or A2 . However, a derivative of this construct containing only a fragment of the Tn5 insertion sequence was able to transform P haemolytica . A further construct containing a fragment of the P haemolytica A1 leucotoxin A gene, was similarly restricted to transforming E coli . These results demonstrate that the pAB2 plasmid is capable of acting as an E coli/P haemolytica shuttle vector . However, the nature of the cloned DNA sequences are important to transformation.

Z Orthop Ihre Grenzgeb, 1995 Mar-Apr, 133(2), 154 - 8
{Bone and joint infections due to unusual pathogens}; Schleicher U et al.; Between 1989 and 1993, 7 patients--2 men and 5 women from 19 to 70 years of age--with osteomyelitis due to an unusual organism were observed . 3 cases with Salmonella typhimurium and one at a time with Escherichia coli, Peptostreptococcus, Propionibacterium acnes and Pasteurella multocida . 5 patients were treated operative accompanied with antibiotics and 1 without operation . 1 patient declined the indicated surgical therapy . The outcome were total healing in 3 cases, healing with ankylosis of the concerning joints in 2 cases and healing with defect in 1 case.

Zentralbl Veterinarmed B, 1995 Mar, 42(1), 28 - 34
Effect of storage on the prevalent alum-precipitated hemorrhagic septicaemia vaccine in Pakistan and preparation of a more efficient oil adjuvant vaccine using dense culture of Pasteurella multocida Roberts type 1 on an improved culture medium; Sheikh MA et al.; Significantly drastic effects of storage on the potency of the alum-precipitated haemorrhagic septicaemia (APHS) vaccine are reported . The APHS vaccine, studied through challenge infection of vaccinated rabbits (post-60 days of vaccination), showed 100% potency when stored at 4 degrees C for 30 days . The potency dropped to 20% when storage period was extended to 60 or more days . At 30 degrees C, the potency reduced by 40, 40 and 60%, respectively, after 30, 60 and 90 days of storage, while, at 37 degrees C, the decrease was 60, 60 and 100% after 30, 60 and 90 days of storage, respectively . In view of this, the oil-adjuvant (OA) HS vaccine was developed by culturing Pasteurella multocida on a medium comprising yeast extract, sucrose, trypticase and sodium bicarbonate, under continuous aeration at 37 degrees C . This gave a far better bacterial count (maximum count 15 x 10(8)/ml) than the conventional APHS vaccine (maximum count 6 x 10(8)/ml) . The OAHS vaccine-carrying water-in-oil emulsion remained stable at room temperature for 1 year . The log protection values of the two batches of the OAHS vaccine, studied in mice, were 5.2 and 5.3, as against 1.9 of the APHS vaccine.

FEBS Lett, 1995 Feb 20, 360(1), 62 - 6
The potent mitogen Pasteurella multocida toxin is highly resistant to proteolysis but becomes susceptible at lysosomal pH; Smyth MG et al.; The susceptibility of the potent mitogen Pasteurella multocida toxin (PMT) to various proteases was investigated . PMT at a toxin to protease molar ratio of 1:1 was resistant to 8 of the 11 proteases tested after one hour . With longer incubation, PMT remained resistant to 7 proteases, and this correlated with a retention of biological activity, indicating that PMT might not require proteolytic cleavage at least until it bound to a cell receptor . Previous evidence had suggested that PMT is processed in the cell via an endosome or lysosome . We have shown that PMT became susceptible to proteolysis when the pH was lowered to 5 or below . This supports the previous suggestion that PMT is processed via a low pH compartment in the cell.

Infect Immun, 1995 Feb, 63(2), 381 - 8
Induction of inflammatory cytokines in bovine alveolar macrophages following stimulation with Pasteurella haemolytica lipopolysaccharide; Yoo HS et al.; Bovine tumor necrosis factor alpha (TNF-alpha) and interleukin-1 beta (IL-1 beta) cDNAs were generated by reverse transcription and then by PCR amplification from lipopolysaccharide (LPS)-stimulated alveolar macrophage RNA . The amplified cDNAs were cloned into pPow and expressed in Escherichia coli DH5 alpha . The expressed proteins were confirmed as TNF-alpha and IL-1 beta by Western blot (immunoblot) analysis and bioassays . We then used the cloned genes as probes in Northern (RNA) blots and investigated the kinetics of TNF-alpha and IL-1 beta mRNA expression in bovine alveolar macrophages stimulated with purified LPS from Pasteurella haemolytica 12296 . The effect of LPS on TNF-alpha and IL-1 beta gene expression was dose dependent, and induction was observed at a concentration of 0.01 microgram/ml . Both TNF-alpha and IL-1 beta mRNA expression were detectable within 0.5 h after stimulation with 1 microgram of LPS per ml, peaked at 1 to 2 h, steadily declined up to 16 h, and were undetectable by 24 h . Secreted TNF-alpha measured by bioassay peaked at 4 h and accumulated at a lesser concentration in conditioned medium throughout the 24 h . By contrast, secreted IL-1 beta was induced at 8 h and reached a maximal concentration at 24 h after stimulation . The ability of LPS to induce TNF-alpha and IL-1 beta gene expression and secretion of bioactive proteins were suppressed by polymyxin B . Our findings support a role for LPS from P . haemolytica in the induction of inflammatory cytokines in bovine pneumonic pasteurellosis.

Mol Microbiol, 1995 Feb, 15(4), 689 - 702
A gene required for nutritional repression of the Bacillus subtilis dipeptide permease operon; Slack FJ et al.; An insertion mutation was isolated that resulted in derepressed expression of the Bacillus subtillis dipeptide transport operon (dpp) during the exponential growth phase in rich medium . DNA flanking the site of insertion was found to encode an operon (codVWXY) of four potential open reading frames (ORFs) . The deduced product of the codV ORF is similar to members of the lambda Int family; CodW and CodX are homologous to HsIV and HsIU, two putative heat-shock proteins from Escherichia coli, and to LapC and LapA, two gene products of unknown function from Pasteurella haemolytica . CodX also shares homology with a family of ATPases, including ClpX, a regulatory subunit of the E . coli ClpP protease . CodY does not have any homologues in the data-bases . The insertion mutation and all previously isolated spontaneous cod mutations were found to map in codY . In-frame deletion mutations in each of the other cod genes revealed that only codY is required for repression of dpp in nutrient-rich medium . The codY mutations partially relieved amino acid repression of the histidine utilization (hut) operon but had no effect on regulation of certain other early stationary phase-induced genes, such as spoVG and gsiA.

Aust Vet J, 1995 Feb, 72(2), 45 - 50
Inflammation and increased numbers of bacteria in the lower respiratory tract of horses within 6 to 12 hours of confinement with the head elevated; Raidal SL et al.; Confinement of horses with their heads elevated for periods up to 24 hours was used to evaluate the extent and the effects of bacterial contamination of the equine lower respiratory tract . Significant (P < 0.05) increases in bacterial numbers (up to 10(9) colony forming units/mL in transtracheal aspirate derived samples) occurred within 6 or 12 hours in most horses . Pasteurella/Actinobacillus spp and Streptococcus spp were most commonly isolated . Lowering of the head for 30 minutes every 6 hours to facilitate postural drainage did not prevent multiplication of organisms to levels equivalent to those achieved by horses where the head was elevated for 24 hours . When horses were released from confinement and heads were no longer maintained in an elevated position, clearance of accumulated secretions and bacteria occurred within 8 to 12 hours . Thus, confinement with the head elevated resulted in significant bacterial contamination and multiplication within the lower respiratory tract during a period often encountered in routine management procedures, such as transportation . The clearance of accumulated secretions occurred over a prolonged period after release from such confinement.

J Clin Microbiol, 1995 Feb, 33(2), 331 - 5
Proposed quality control guidelines for antimicrobial susceptibility tests using tilmicosin; Shryock TR et al.; Quality control guidelines for tilmicosin, a novel veterinary-use-only macrolide, were developed in a multi-laboratory study according to established National Committee for Clinical Laboratory Standards (NCCLS) procedures (M23-T2) . Tilmicosin was incorporated into Sensititre plates for broth microdilution endpoint testing and into two lots of 15-micrograms disks for Kirby-Bauer agar disk diffusion testing . One common lot and five unique lots of Mueller-Hinton media were used . (Broth was cation adjusted, and agar was supplemented with 5% defibrinated sheep blood.) Bacteria used for reference strains included Pasteurella haemolytica 128K, Pasteurella multocida ATCC 43137, and Staphylococcus aureus ATCC 29213 (microdilution) and ATCC 25923 (disk) . Replicate tests were conducted . Disk diffusion and broth microdilution quality control ranges are proposed.

Microbios, 1995, 83(336), 161 - 6
Detection and partial characterization of Pasteurella multocida found in feline skin lesions; Hoshuyama S et al.; Pasteurella multocida was isolated from 21 of 105 purulent skin lesions in household cats . The bacterium was in pure culture in nine specimens and predominant in six specimens . Its viable counts were 10(2) to 10(7) colony forming units/ml . Of 21 isolates of P . multocida, seventeen were considered to be capsular type A . The predominant capsular and somatic type was the serotype A:3,4 . Inoculation of the filter-sterilized supernatants of the isolates induced an erythematous response in guinea pig skins . These findings suggest that P . multocida is a candidate as a pathogen of feline skin lesions and the erythema-inducing activity of the bacterium may participate in the formation of skin lesions in household cats.

Drugs, 1995, 49 Suppl 2, 152 - 8
Veterinary use of new quinolones in Japan; Nakamura S; Five new quinolones are used as veterinary medicines in Japan . Benofloxacin and ofloxacin are orally given in feed or in drinking water for respiratory diseases due to Mycoplasma spp . and Escherichia coli in poultry . Enrofloxacin is subcutaneously, intramuscularly or orally administered to cattle and swine for pneumonia due to Pasteurella spp., Mycoplasma spp . and Actinobacillus pleuropneumoniae and diarrhoea due to E . coli, and is used orally for respiratory diseases in poultry . Danofloxacin is similar to enrofloxacin except that the former is not approved for diarrhoea in cattle and swine . Orbifloxacin is given intramuscularly for pneumonia and diarrhoea in cattle and swine but not for poultry diseases . They are effective and safe in animals, and disappear from edible tissues after appropriate drug withdrawal times . The quinolones are indicated only when first-choice drugs are ineffective, only under the direction of veterinarians and only for periods of less than 5 days . Under these conditions, it appears unlikely that quinolone use in veterinary practice will be detrimental to human chemotherapy.

J Antibiot (Tokyo), 1995 Jan, 48(1), 67 - 72
Effect of polymyxin B nonapeptide on daptomycin permeability and cell surface properties in Pseudomonas aeruginosa, Escherichia coli, and Pasteurella multocida; Morris CM et al.; The present study was carried out to determine if sensitization of Gram-negative bacteria to the polyanionic antibiotic daptomycin by cationic molecules can be explained on the basis of decreased cell surface charge in order to better understand intrinsic resistance . Turbidimetric assessments of batch cultural growth kinetics revealed the outer membrane permeabilizer polymyxin B nonapeptide sensitized Pseudomonas aeruginosa and Escherichia coli to the hydrophobic probe novobiocin, whereas little or no sensitization was observed for two surface hydrophobicity variants of Pasteurella multocida . Polymyxin B nonapeptide and daptomycin synergistically inhibited growth of P . aeruginosa only . A hydrocarbon adherence assay revealed permeabilizing concentrations of polymyxin B nonapeptide increased cell surface hydrophobicity of P . aeruginosa and the hydrophobic P . multocida variant, while E . coli and the hydrophilic P . multocida variant remained unaffected . Measurement of cellular electrophoretic mobility showed polymyxin B nonapeptide permeabilization of P . aeruginosa to daptomycin occurred concomitantly with a significant decrease in cell surface charge, while no such sensitization occurred in organisms which failed to undergo polymyxin B nonapeptide-induced surface charge decreases . These data suggest that sensitization of Gram-negative bacteria to polyanionic lipopeptides by growth in the presence of polycationic outer membrane permeabilizers such as polymyxin B nonapeptide is dependent on diminution of overall cell surface charge and polarity, thereby allowing outer cell envelope permeation.

Avian Dis, 1995 Jan-Mar, 39(1), 94 - 9
Serotypes and DNA fingerprint profiles of Pasteurella multocida isolated from raptors; Wilson MA et al.; Pasteurella multocida isolates from 21 raptors were examined by DNA fingerprint profile and serotyping methods . Isolates were obtained from noncaptive birds of prey found in 11 states from November 28, 1979, through February 10, 1993 . Nine isolates were from bald eagles, and the remaining isolates were from hawks, falcons, and owls . Seven isolates were members of capsule group A, and 14 were nonencapsulated . One isolate was identified as somatic type 3, and another was type 3,4,7; both had unique HhaI DNA fingerprint profiles . Nineteen isolates expressed somatic type 1 antigen; HhaI profiles of all type 1 isolates were identical to each other and to the HhaI profile of the reference somatic type 1, strain X-73 . The 19 type 1 isolates were differentiated by sequential digestion of DNA with HpaII; four HpaII fingerprint profiles were obtained . The HpaII profile of one isolate was identical to the HpaII profile of strain X-73 . Incidence of P . multocida somatic type 1 in raptors suggests that this type may be prevalent in other wildlife or wildlife environments.

Avian Dis, 1995 Jan-Mar, 39(1), 145 - 6
Incidence of Pasteurella multocida in poultry house cats used for rodent control programs; Van Sambeek F et al.; Cats used as mousers in commercial poultry breeder farms were tested for the presence of Pasteurella multocida . Of the cats tested, 72.7% were positive for P . multocida serotypes common to both farm cats and commercial broiler breeders . Thirty-one percent were type 1, 19% were type 4, and 6% were type 3.4.

Avian Dis, 1995 Jan-Mar, 39(1), 141 - 4
Consumptive coagulopathy in turkeys exposed to Pasteurella multocida; Friedlander RC et al.; When turkeys were inoculated intramuscularly with live Pasteurella multocida, three of the four inoculated turkeys developed an increase in modified Russell's viper venom time (mRVVT) 24 hours after inoculation . This increase was followed by irregular decreases and increases in mRVVT at subsequent bleedings . When turkeys were inoculated intravenously with P . multocida, the mRVVT increased markedly after inoculation in all eight inoculated turkeys: 9 hours later, the average mRVVT was significantly (P < 0.05) higher than that of the uninoculated turkeys . No microthrombi were observed in the blood vessels of the liver, spleen, kidneys, or lungs . An increase in mRVVT was interpreted as an excessive consumption of one or more of clotting factors X, V, II, and I . These results indicate that consumptive coagulopathy could be a factor in the pathogenesis of fowl cholera in turkeys.

Protein Sci, 1995 Jan, 4(1), 126 - 9
Identification of a eukaryotic-like protein kinase gene in Archaebacteria; Smith RF et al.; Primary sequence patterns based on known conserved sites in eukaryotic protein kinases were used to search for eukaryotic-like protein kinase sequences in a six-frame translation of the bacterial subsection of GenBank . This search identified a previously unrecognized eukaryotic-like protein kinase gene in three related methanogenic archaebacteria, Methanococcus vannielii, M . voltae, and M . thermolithotrophicus . The proposed coding sequences are located in orthologous open reading frames (ORFs): ORF547, ORF294, and ORF114, respectively . The C-terminus of the ORFs contains 9 of the 11 subdomains characteristically conserved within the eukaryotic protein kinase catalytic domain . The N-terminus of the ORFs is similar to a putative glycoprotease in Pasteurella haemolytica and its homologue in Escherichia coli, the orfX gene . This is the first report of a eukaryotic-like protein kinase sequence observed in Archaebacteria.

Dermatology, 1995, 190(2), 145 - 9
Necrotizing fasciitis due to Pasteurella multocida infection; Hamamoto Y et al.; Necrotizing fasciitis is a potentially fatal clinical disease caused by infection with various bacteria in addition to streptococci, which are common causative agents . We report on a rare case of this disease in association with Pasteurella multocida infection . A 58-year-old man had systemic features of shock after a 15-hour history of a painful swelling on the right lower leg . The swelling led to skin blistering and necrosis from which P . multocida was isolated . Those lesions progressed rapidly . The patient also had a history of chronic liver injury as described in previous reports.

Res Vet Sci . 1995 Jan;58(1):98.
Characterisation of a new Pasteurella haemolytica serotype (A17); Younan M et al.; Four strains of Pasteurella haemolytica representing a new serotype (A17) were isolated from sheep in Syria . The identity of the new serotype was established by comparison with P haemolytica type strains A1 to A16 in the indirect haemagglutination test.

Lab Anim, 1995 Jan, 29(1), 59 - 65
An enzyme-linked immunosorbent assay (ELISA) for monitoring antibodies to SP group Pasteurellaceae in guineapigs; Boot R et al.; Cross-reactivity studies with Pasteurellaceae from guineapigs revealed 5 serologically distinct groups, comprising Pasteurella multocida, Sp group bacteria, SP-like bacteria, Pasteurella pneumotropica and an actinobacillus-like bacterium . Guineapig Pasteurellaceae differed serologically from mouse-derived P . pneumotropica NCTC 8284 . An enzyme-linked immunosorbent assay (ELISA) using SP group antigen, developed to monitor 'natural' infections by SP group Pasteurellaceae in guineapigs, detected significantly more infection than did cultivation, and was found superior to an ELISA performed with P . pneumotropica NCTC 8284.

Can J Vet Res, 1995 Jan, 59(1), 46 - 50
Expression of iron-regulated outer membrane proteins by porcine strains of Pasteurella multocida; Zhao G et al.; The outer membrane protein (OMP) profiles of two strains of capsular type A Pasteurella multocida isolated from the lungs of pigs with enzootic pneumonia were studied . Sarkosyl extracted OMPs from P . multocida grown under iron-restricted and iron-replete conditions were subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blot analysis . Results showed that the iron-regulated outer membrane proteins (IROMPs) with molecular masses of 74 kDa, 94 kDa, 99 kDa and 109 kDa were expressed by strain A52, while 74 kDa, 82 kDa, 94 kDa and 99 kDa IROMPs were expressed by strain B80 . Swine immune sera, obtained from pigs which were first immunized with a polyvalent P . multocida type A and type D bacterin and subsequently challenged with type A strain of P . multocida, contained antibodies against the IROMPs . These antibodies cross-reacted with the IROMPs expressed by avian strain P1059 of P . multocida . Convalescent-phase serum obtained from turkeys which survived fowl cholera, also cross-reacted with the IROMPs from porcine strains of P . multocida . These results suggested that IROMPs from porcine and avian strains of P . multocida may share common epitopes that were recognized by swine immune serum as well as turkey convalescent-phase serum.

J Clin Microbiol, 1995 Jan, 33(1), 202 - 4
Pasteurella multocida septicemia and subsequent Pasteurella dagmatis septicemia in a diabetic patient; Fajfar-Whetstone CJ et al.; Pasteurella species may cause zoonotic infections of humans . Serious systemic infections with these organisms are unusual, but they may occur in individuals with predisposing underlying illnesses . Occurrences of bacteremia due to P . multocida are infrequent, and P . dagmatis bacteremia is even rarer . We report independent occurrences of P . multocida and P . dagmatis septicemia in the same diabetic patient after contact with two pet dogs . We review the history of Pasteurella species and discuss the biochemical and clinical features of its association with zoonosis.

Mycoses, 1995 Jan-Feb, 38(1-2), 85 - 91
Studies of the influence of ochratoxin A on immune and defense reactions in the mouse model; Muller G et al.; In the mouse model, the mycotoxin ochratoxin A has a non-selective suppressive effect on various immune and defence reactions . Apart from weight depression, lymphopenia, neutrophilia and eosinophilia, antibody-producing cells, antibody titres in blood serum and phagocytosis of Escherichia coli by blood phagocytes become suppressed . Moreover, immunized animals show a lower survival rate after experimental infection with Pasteurella multocida as well as an increase in oxygen radicals in blood cells.

DNA Seq, 1995, 5(5), 291 - 7
Sequence analysis of leukotoxin secretion determinants from a Pasteurella haemolytica-like organism; Chang YF et al.; The pHLBD genes encoding the secretion functions for the 105 kDa RTX leukotoxin of Pasteurella haemolytica-like (PHL) organism has been cloned and sequenced . Like analogous genes from other RTX determinants, the pHLBD genes lie immediately downstream from the leukotoxin structural gene, pHLA . Although isolated from a diverse group of gram-negative organisms, the pHLBD genes and the characterized RTX BD genes from other organisms exhibit a high degree of homology at both the DNA and predicted amino acid sequence levels . We have previously reported the cloning of the leukotoxin gene (pHLCA) (Chang et al., Infect . Immun . 61:2089-2095), which encodes a 105-kda polypeptide with cytotoxic activity . DNA sequence analysis of the pHLBD genes shows 83.93% and 86.05% homologous to that of P . haemolytica IktBD genes, respectively.

J Wildl Dis, 1995 Jan, 31(1), 22 - 9
Molecular characterization of Pasteurella testudinis isolated from desert tortoises (Gopherus agassizii) with and without upper respiratory tract disease; Snipes KP et al.; Isolates of Pasteurella testudinis recovered from clinically healthy desert tortoises (Gopherus agassizii) and tortoises with upper respiratory tract disease (URTD) were characterized in an attempt to identify strains associated with disease . Eighty-nine isolates, 52 from ill and 37 from healthy tortoises collected from Nevada (USA), June 1990 to September 1991, were genomically fingerprinted and grouped based on ribotype similarity . Twelve isolates (six from ill and six from healthy tortoises) were further characterized with regard to whole-cell protein (WCP) and outer membrane protein (OMP) composition and their ability to survive in normal tortoise plasma . The 89 isolates were initially distributed into 33 distinct ribotype groups using the restriction enzyme EcoRI; five ribotypes contained over 50% of the isolates . Only one EcoRI ribotype was comprised of multiple isolates (n = 4) exclusively recovered from tortoises with URTD . When the ten EcoRI ribotypes that contained more than one isolate per ribotype were further studied using a second restriction enzyme, EcoRV, one EcoRI/EcoRV ribotype contained five isolates recovered from URTD tortoises and none from healthy animals . The EcoRI ribotype comprised of four isolates, all from tortoises with URTD, was further separated into three distinct groups with EcoRV . All 12 isolates studied grew equally well in normal tortoise plasma, and when broth-grown WCP and OMP profiles were evaluated, no proteins were unique to isolates from URTD tortoises . Iron-regulated OMP's were produced in three isolates examined, but these OMP's apparently were not virulence-related.

Int J Syst Bacteriol, 1995 Jan, 45(1), 139 - 44
Small-subunit rRNA sequences and whole DNA relatedness concur for the reassignment of Pasteurella piscicida (Snieszko et al.) Janssen and Surgalla to the genus Photobacterium as Photobacterium damsela subsp . piscicida comb . nov; Gauthier G et al.; The taxonomic status of Pasteurella piscicida (strain NCIMB 2058T {T = type strain} and a strain isolated from the environment) was investigated by performing phylogenetic analyses of small-subunit rRNA sequences, DNA-DNA hybridization analyses, and biochemical characterization analyses . The results of the phylogenetic analyses and the levels of DNA-DNA complementarity demonstrated conclusively that Pasteurella piscicida is extremely closely related to Photobacterium damsela ATCC 33539T . Since the two taxa exhibited a level of DNA-DNA relatedness of 80%, they are members of the same species . The high level of DNA relatedness and the presence of specific morphological and biochemical characteristics support the hypothesis that two subspecies should be recognized . On the basis of its phylogenetic position, we concluded that Pasteurella piscicida should be renamed Photobacterium damsela subsp . piscicida comb . nov.

FEMS Microbiol Lett, 1994 Dec 15, 124(3), 285 - 9
Capsular polysaccharide expressed by Pasteurella piscicida grown in vitro; Bonet R et al.; Pasteurella piscicida grown in a glucose-rich medium produces a capsule that can be see under light and electron microscopy . The capsular polysaccharide was purified and characterized by chemical and HPLC analysis . The polymer has the composition glucose/mannose/N-acetylgalactosamine/galacturonic acid/acetic acid in the molar ratios of approximately 2.5:1.3:0.5:0.4:2.5 . The polysaccharide was immunogenic in rabbits and did not cross-react with antibodies against the O-antigen lipopolysaccharide.

J Biol Chem, 1994 Dec 9, 269(49), 31289 - 95
Identification and immunological characterization of the domain of Actinobacillus actinomycetemcomitans leukotoxin that determines its specificity for human target cells; Lally ET et al.; Although extensive amino acid homology exists among the various Repeats in ToXin (RTX) family of bacterial cytolysins, the cellular and species specificities remain unique for individual toxins (i.e . Actinobacillus actinomycetemcomitans leukotoxin (LtxA) kills human monomyelocytes while a related toxin, Pasteurella hemeolytica leukotoxin (LktA) kills bovine lymphoid cells) . To determine the Ltx domain responsible for species specificity, ltxA/lktA chimeric toxin genes were expressed in tandem with the ltxC gene under control of the P lambda promoter . The ability of lysates to kill either HL-60 (human) or BL-3 (bovine) cells was assessed by trypan blue exclusion . The critical area required for the chimeric toxins to recognize human target cells is a 253-amino acid fragment (residue 688-941) that contains the GGXGXDX(L{I{V{W{Y{F)X repeats . A panel of 12 neutralizing anti-LtxA monoclonal antibodies also recognized specificities within the 253-amino acid fragment . Epitope mapping of the monoclonal antibody panel showed that all antibodies bound to one of three sites on the LtxA molecule . One monoclonal recognized epitope A which was composed of LtxA residues 698-709 (KLDYYYTNKGFK), six antibodies recognized epitope B, a peptide composed of residues 746-757 (LIYGYDGDDRLY), whereas the remaining five monoclonals recognized epitope C, which is composed of residues 926-937 (DRARLKRQFELQ).

Am J Vet Res, 1994 Dec, 55(12), 1703 - 9
Characterization of an Actinobacillus pleuropneumoniae seeder pig challenge-exposure model; Lechtenberg KF et al.; Five strains of Actinobacillus pleuropneumoniae serotype 1 were used to intranasally infect 5 groups of pigs . Using each bacterial strain, infected pigs (termed seeder pigs) were commingled for 48 hours with 5 groups of noninfected test pigs, then were removed . Seeder and test pigs were maintained in isolation and were observed for 14 days . Seeder pigs had mortality that was threefold greater than that of test pigs (24% vs 8%) . Rectal temperature in excess of 40.3 C was achieved for 84% of test pigs and 88% of seeder pigs . Neither of these 2 variables was statistically different between the 2 groups of pigs . Clinical impression scores > or = 2 (on a 0 to 3 scale) were three-fold (64% vs 20%) greater for seeder than for test pigs (P < 0.05) . The total number of bacterial isolations or nonrecoverable isolates was tabulated for test and seeder pigs' lungs at necropsy, irrespective of the amount of lesions . The number of A pleuropneumoniae isolations was not statistically different between test and seeder pig populations . Recovery of Pasteurella multocida or other bacteria was greater from the seeder pigs (P < 0.05), whereas the number of non-recoverable isolates was greater from test pigs than from seeder pigs (P < 0.05) . Assessment of lung lesions at necropsy by either visual estimation or on a weight basis were in agreement.(ABSTRACT TRUNCATED AT 250 WORDS)

Microbiology, 1994 Dec, 140 ( Pt 12), 3293 - 300
Comparison of outer-membrane proteins of Pasteurella haemolytica expressed in vitro and in vivo in cattle; Davies RL et al.; Outer-membrane protein (OMP) profiles of two serotype A1 isolates of Pasteurella haemolytica were compared by SDS-PAGE and Western blotting with bovine convalescent serum after growth (a) in vitro under iron-sufficient and -deficient conditions, (b) in vivo in the lungs of experimentally infected calves and (c) in vivo in diffusion chambers implanted into the peritoneal cavities of calves . Lung-grown bacteria differed from iron-sufficient in vitro-grown bacteria in having enhanced expression of the previously recognized 71, 77 and 100 kDa iron-regulated proteins, reduced expression of 18, 31, 39.5 and 50 kDa proteins, and expression of a 19 kDa protein . Differences were also apparent in the Western blot profiles of OMPs of in vitro- and lung-grown bacteria . These included the apparent lack of recognition of the 100 kDa protein in the lung-grown bacteria, but not in the in vitro-grown bacteria, and more intense staining of a 47 kDa protein in in vitro-grown bacteria, but not in lung-grown bacteria . The OMP profiles of the chamber-grown bacteria resembled those of the lung-grown bacteria in that expression of the 18, 19, 31 and 39.5 kDa proteins was similar . These similarities demonstrated that the chamber-grown bacteria had adapted to the in vivo environment, and that growth conditions within the chambers resembled, but not perfectly, those within the lungs . For example, expression of the three iron-regulated OMPs was very low in the chamber-grown bacteria compared to the lung-grown bacteria.(ABSTRACT TRUNCATED AT 250 WORDS)

Tierarztl Prax, 1994 Dec, 22(6), 524 - 8
{Clinical, diagnostic laboratory and therapeutic studies of mastitis in a large sheep breeding flock}; Bocklisch H et al.; During two lambing seasons, 6500 ewes of a large sheep breeding unit were investigated . Within this period, 467 ewes suffered from mastitis . The morbidity was 3.8% and the lethality 4.7% . In 84% of the investigated animals udder-pathogenic bacteria were detected . Mostly, Pasteurella haemolytica and Staphylococcus aureus were isolated as agents of ovine mastitis . Before and during the antibiotic therapy the clinical symptoms (general condition, temperature, milk appearance, consistency of the udder tissue) were recorded . The combined antibiotic therapy, which consisted of parenteral and local intracisternal applications, was mostly satisfactory concerning recovery from mastitis and survival of the ewes . Only 17 ewes died from mastitis; most deaths caused by Staphylococcus aureus.

J Leukoc Biol, 1994 Nov, 56(5), 644 - 9
Pasteurella haemolytica leukotoxin-induced synthesis of eicosanoids by bovine neutrophils in vitro; Clinkenbeard KD et al.; Exposure of isolated bovine neutrophils to partially purified Pasteurella haemolytica leukotoxin caused increased synthesis of leukotriene B4 (LTB4) but not thromboxane B2 (TXB2) from endogenous arachidonic acid . Synthesis of LTB4 was closely correlated with leukotoxin-induced neutrophil lysis . At low toxin concentrations, LTB4 production lagged behind leukotoxin-induced neutrophil lysis over a 3-h period . The neutralizing monoclonal antileukotoxin antibody MM601 neutralized both leukotoxin-induced neutrophil lysis and LTB4 synthesis . Both leukotoxin-induced neutrophil lysis and LTB4 synthesis were Ca(2+)-dependent . When leukotoxin-induced LTB4 synthesis from exogenous arachidonic acid was examined, significant LTB4 synthesis occurred at 5 min of leukotoxin exposure, which was before leukotoxin-induced lysis developed . Leukotoxin-induced LTB4 synthesis from endogenous arachidonic acid appears to require leukotoxin-induced plasma membrane damage (occurring during neutrophil lysis), whereas LTB4 synthesis from exogenous arachidonic acid is initiated rapidly and occurs in the absence of plasma membrane damage.

Dtsch Tierarztl Wochenschr, 1994 Nov, 101(11), 441 - 3
Studies of the pathology and microbiology of pneumonic lungs of lambs; Haziroglu R et al.; The characteristics of lung lesions of slaughtered lambs and the presence of microorganisms in the lesions were investigated . Pneumonia was detected in 500 (3.6%) of 13,588 lambs . Macroscopical lesions were of atypical pneumonia and were often seen in the pars cranialis of lobus cranialis dexter . Histologically, proliferative pneumonia was detected in most cases and was often accompanied by exudative characteristics . Pasteurella haemolytica was isolated from 258 (51.6%) and Mycoplasma ovipneumoniae from 215 (43.0%) of 500 pneumonic lungs . In 131 (26.2%) cases both organisms were isolated from same samples . A close relationship was found between P . haemolytica and exudative inflammation (p < 0.05).

Clin Infect Dis, 1994 Nov, 19(5), 941 - 3
Aortic graft infection due to Pasteurella haemolytica and group C beta-hemolytic streptococcus; Rivera M et al.; Although infection is a rare complication of aortic bypass grafting, we treated a 50-year-old patient who developed aortic graft infection due to Pasteurella haemolytica and group C beta-hemolytic streptococcus . The source of the infection could not be verified; however, after removal of the infected graft and administration of a 6-week course of intravenous ampicillin, he recovered fully . We discuss the etiology and pathogenesis of this rare infection.

Med Microbiol Immunol (Berl), 1994 Nov, 183(5), 229 - 37
The characterization of ion channels formed by Pasteurella multocida dermonecrotic toxin; Krasilnikov OV et al.; The influence of the dermonecrotic lethal toxin (approximately 120 kDa) produced by Pasteurella multocida serovarian D on planar phospholipid bilayers was studied . It was found that the toxin is able to increase the conductance of the bilayers by formation of low-conductive and cation-selective ion channels {27 pS at 4.0 M KCl, pH 7.5; zero current potential equals to -14.5 +/- 0.5 mV at threefold transmembrane gradient KCl (120 mM/40 mM)} . In biionic conditions the channels displayed weak selectivity between Na, K and Ca ions . The shapes of current-voltage characteristics (which were measured at different pH and salt concentrations) indicate that an energetic barrier for passing ions is situated near the center of the water pore of the ion channels . The effective diameter of the ion channel's water pore was established to be equal to 2.1 +/- 0.3 nm.

Gene, 1994 Oct 11, 148(1), 101 - 5
Construction of isogenic mutants of Pasteurella haemolytica by allelic replacement; Murphy GL et al.; We describe methods for the mutagenesis of cloned Pasteurella haemolytica (Gram-) genes and for the construction of P . haemolytica mutants by allelic exchange . We used these methods to construct isogenic mutants of P . haemolytica which no longer synthesize three membrane lipoproteins (Lpp) . A single genetic locus, consisting of three tandemly arranged genes encoding 28-30-kDa membrane Lpp, was replaced with a mutated locus which carries the beta-lactamase-encoding ApR gene from a 4.2-kb P . haemolytica plasmid . The inactivated locus was introduced into P . haemolytica by electroporation of a plasmid which carries the mutated locus, but is incapable of replicating in P . haemolytica . Southern and Western blot analyses indicate that the wild-type locus was replaced by the mutated locus through a double-crossover recombination event and that the membrane Lpp were no longer produced by the mutant strain . These methods should be useful in constructing mutant loci which can be used to analyze the roles for various P . haemolytica proteins in the pathogenesis of bovine pneumonic pasteurellosis.

Infect Immun, 1994 Oct, 62(10), 4675 - 8
In vivo production of neuraminidase by Pasteurella haemolytica A1 in goats after transthoracic challenge; Straus DC et al.; Nine goats were injected transthoracically with Pasteurella haemolytica A1 to determine if an extracellular bacterial enzyme, neuraminidase, was produced in vivo during infection with this organism . The principal group of goats (n = 9) each received 1 ml of 7.25 x 10(5) live P . haemolytica A1 cells in polyacrylate beads transthoracically in the left lung on days 0 and 21 . Six goats were used as negative controls and received 0.3 g of polyacrylate beads subcutaneously in the right flank on days 0 and 21 . Serum was obtained from all animals on days -4, 3, 7, 14, 21, 24, and 32 . Preimmune serum from all animals showed no detectable antibody to P . haemolytica A1 neuraminidase in an enzyme neutralization assay . None of the sera from the negative control animals possessed a significant antibody concentration in response to the P . haemolytica A1 neuraminidase . On day 32, serum samples from the nine infected animals possessed enzyme neutralizing activity that ranged from 62% to 100% . Anti-neuraminidase antibody could be detected as early as day 14 by the enzyme neutralization assay . These data demonstrate that the enzyme neuraminidase is produced in vivo during an active P . haemolytica A1 lobar infection.

Infect Immun, 1994 Oct, 62(10), 4580 - 6
Mycoplasma agassizii causes upper respiratory tract disease in the desert tortoise; Brown MB et al.; The desert tortoise is listed by the United States government as a threatened species in part of its range . A major contributing factor in the decline of this animal has been the presence of an upper respiratory tract disease (URTD) which is characterized by a chronic disease which eventually leads to severe occlusion of the nares with viscous exudate and destruction of the respiratory epithelium . Electron microscopy of infected tissues demonstrated the presence of a mycoplasma-like organism attached to the respiratory surfaces . The mycoplasma was isolated and designated as a new species, with the proposed name Mycoplasma agassizii . The current study was designed to fulfill Koch's postulates and determine if M . agassizii was the etiologic agent of URTD . Clinically healthy animals with known antibody status were infused intranasally with pooled exudate (n = 8) from ill donor animals, with M . agassizii alone (n = 9) or in combination with Pasteurella testudinis (n = 8), with P . testudinis alone (n = 9), or with sterile broth (n = 12) . The pooled exudate was culture positive for M . agassizii . Tortoises which received exudate or M . agassizii alone or in conjunction with P . testudinis were significantly more likely to develop clinical disease (P < 0.0004) than animals which received P . testudinis alone or the broth controls . Tortoises demonstrated a strong immune response to M . agassizii, and seroconversion was seen in all groups with clinical disease . M . agassizii was isolated from the upper respiratory tracts of clinically ill animals up to 6 months postinfection . On the basis of the results of these transmission studies, we conclude that M . agassizii is an etiologic agent of URTD in the desert tortoise.

Vet Q, 1994 Oct, 16(3), 148 - 52
Double blind field evaluation of a trivalent vaccine against respiratory disease in veal calves; Frankena K et al.; A field trial was performed to determine the effect of a trivalent vaccine on Clinical Respiratory Tract Problems (CRTP) in veal calves . The vaccine has been developed to increase immunity against the causal agents of IBR, BVD and BRSV infection . In total 928 calves, housed in 16 compartments of one herd, were involved . In four compartments of 58 calves each, vaccinated and non-vaccinated calves were housed together . Four other compartments were treated as a whole and 8 compartments were left untreated . CRTP incidence, medications, weight gain, haemoglobulin and IgG level were recorded . From CRTP positive animals, seroconversion and presence of specific bacteriae and/or viruses were measured as well . Results of the compartments where vaccinated and non-vaccinated calves were housed together showed that the incidence of CRTP in vaccinated calves was 0.16 while it amounted to 0.28 in controls . Most cases were found between day 40 and 60 after the start of the trial . Seroconversion for vaccine specific viruses was sporadically found, but the presence of Pasteurella's was confirmed in the majority of cases . Presumably, the higher incidence of CRTP in the control group was due to a higher level of BVDV infection which might facilitate a clinical infection with Pasteurella's . Vaccination was also negatively related to the percentage of affected lungs at slaughter, the number of days antibiotics had to be administered and subsequently to medicine costs, although these effects were not significant . Daily weight gain was significantly affected by CRTP, but not by vaccination . The effects of vaccination in the compartments where calves were either all vaccinated or not-vaccinated, were similar or larger when compared to the effects in compartments where half of the calves were vaccinated.

Diagn Microbiol Infect Dis, 1994 Oct, 20(2), 105 - 7
Actinobacillus (Pasteurella) ureae meningitis in a HIV-positive patient; Kaka S et al.; Actinobacillus (Pasteurella) ureae is a commensal of the human nasopharynx that is a rare cause of meningitis . We present an HIV-infected patient with this condition . His case, and those previously reported in the literature, implicate head trauma or a neurosurgical procedure in the pathogenesis of this condition . The patient responded to initial empiric therapy with ceftriaxone, followed by intravenous penicillin.

J Vet Med Sci, 1994 Oct, 56(5), 917 - 21
Antibacterial activity of tilmicosin against Pasteurella multocida and Actinobacillus pleuropneumoniae isolated from pneumonic lesions in swine; Inamoto T et al.; Sixty one strains of Pasteurella multocida and 35 strains of Actinobacillus pleuropneumoniae isolated from pneumonic lesions of porcine lungs during the period from 1985 to 1989 in Japan were tested for antibiotic susceptibility to chlortetracycline (CTC), thiamphenicol (TP), tylosin (TS), acetylisovaleryl-tylosin (AIV-TS), tilmicosin (TMS), mirosamycin (MRM) . Most strains of both species were sensitive to CTC, TP and TMS . Growth of fifty-one strains (83.6%) and forty-six strains (75.4%) of P . multocida were inhibited with 3.13 micrograms/ml of CTC and 0.78 micrograms/ml of TP, respectively . TS showed low activity against almost all strains (MIC > or = 6.25 micrograms/ml) . Fifty-eight (95.1%), twenty-three (37.7%) and fifty (82%) of P . multocida showed MICs of > or = 6.25 micrograms/ml against AIV-TS, TMS and MRM, respectively . The MICs of A . pleuropneumoniae against CTC were less than 1.56 micrograms/ml . Thirty-two strains (91.4%) and 33 strains (94.3%) of A . pleuropneumoniae were inhibited with 3.13 micrograms/ml of TP and TMS respectively . However, TS, AIV-TS and MRM showed low activity against all of A . pleuropneumoniae (MIC > or = 6.25 micrograms/ml) . Three different resistance patterns were observed in P . multocida and two in A . pleuropneumoniae isolates, respectively.

Antimicrob Agents Chemother, 1994 Oct, 38(10), 2392 - 7
Effects of localized Pasteurella haemolytica infection on erythromycin-binding properties of bovine alpha-1-acid glycoprotein, albumin, serum, and tissue chamber fluids; Walker JL et al.; The in vitro erythromycin-binding properties of bovine alpha-1-acid glycoprotein (AAG) and albumin were studied by using equilibrium dialysis . In addition, the proportions of free erythromycin in bovine serum and tissue chamber fluid before and 4 days after inoculation of subcutaneous tissue chambers with Pasteurella haemolytica were measured . At a concentration of 5 micrograms/ml, erythromycin was moderately bound to AAG (39% +/- 4% free) and was only slightly bound to albumin (86% +/- 2% free) . Scatchard analysis of the data describing binding to pure bovine AAG indicated that erythromycin was bound to a single high-affinity (6.45 x 10(4) M-1) site on the protein . At lower total concentrations of erythromycin, the free concentrations of the antibiotic were lower in serum samples collected after infection (49% +/- 3% at 5 micrograms of erythromycin per ml) than in those collected before inoculation (55% +/- 3% at 5 micrograms of erythromycin per ml) . Inoculation had no effect on binding to macromolecules in chamber fluids . Inoculated tissue chambers served as a convenient model for studying the effect of infection on drug-macromolecule interactions in interstitial fluid.

J Wildl Dis, 1994 Oct, 30(4), 529 - 35
Susceptibility of phagocytes from elk, deer, bighorn sheep, and domestic sheep to Pasteurella haemolytica cytotoxins; Silflow RM et al.; Alveolar macrophages and peripheral blood neutrophils from elk (Cervus elaphus), bighorn sheep (Ovis canadensis canadensis), and domestic sheep were exposed to culture supernatants from Pasteurella haemolytica isolated from bighorn sheep and domestic sheep . In a second experiment, peripheral blood neutrophils from mule deer (Odocoileus hemionus), elk, and bighorn sheep were exposed to culture supernatants from P . haemolytica isolated from elk, bighorn sheep and domestic sheep . Alveolar macrophages from elk, bighorn sheep and domestic sheep were resistant to killing by P . haemolytica supernatants from bighorn sheep and domestic sheep; susceptibility of neutrophils to cell death, as measured by release of lactate dehydrogenase, differed significantly (P < 0.05) between the four species tested . Bighorn sheep and domestic sheep neutrophils were susceptible to cytotoxin damage by the P . haemolytica isolates used; bighorn sheep neutrophils were four- to eight-fold more susceptible to cytotoxin damage than domestic sheep neutrophils . Neutrophils from deer and elk were resistant to killing by P . haemolytica cytotoxins from any species tested.

J Wildl Dis, 1994 Oct, 30(4), 523 - 8
Comparative leukotoxicities of Pasteurella haemolytica isolates from domestic sheep and free-ranging bighorn sheep (Ovis canadensis); Sweeney SJ et al.; Twenty-eight isolates of Pasteurella haemolytica from domestic sheep (n = 14 isolates) and bighorn sheep (n = 14 isolates) were evaluated for leucotoxicity against peripheral blood neutrophils of bighorn sheep by adding bacterial culture supernatants to bighorn sheep neutrophils in vitro . Leukotoxic isolates of P . haemolytica, defined as causing > 50% neutrophil death as measured by release of lactate dehydrogenase into culture supernatants, were identified from eight of 14 domestic sheep isolates and from 0 of 14 bighorn sheep isolates . The in vitro assay of isolates of P . haemolytica may provide a valid predictive measure of strain virulence of P . haemolytica, and of potential pneumonic episodes in bighorn sheep populations.

Avian Dis, 1994 Oct-Dec, 38(4), 790 - 6
Lesions resulting from attempted Shwartzman reaction in turkey poults inoculated with Pasteurella multocida lipopolysaccharide; Mendes S et al.; Five-week-old turkey poults were given two consecutive intravenous injections (24 hours apart) of highly purified Pasteurella multocida lipopolysaccharide (LPS) in an effort to induce a generalized Shwartzman reaction . There were no gross lesions, and microscopic lesions were limited to focal hepatic necrosis with heterophil infiltration . Hepatic lesions did not differ qualitatively from lesions in turkeys given a single dose of lipopolysaccharide . Margination of heterophils in the pulmonary vasculature was observed in turkeys 4 hours after a single injection of LPS, but it was not present in turkeys given the consecutive injections of LPS . To induce a dermal Shwartzman reaction, turkeys were given intradermal injections of LPS followed by an intravenous injection of LPS 24 hours later . Although no grossly visible hemorrhagic dermal necrosis occurred, microscopic lesions, including heterophil infiltration, vasculitis, thrombosis, and necrosis, were present . Thrombosis and vasculitis were observed only in turkeys given the intravenous and intradermal LPS, whereas the other inflammatory changes were observed in turkeys given the intradermal injection of LPS and intravenous water . Prominent lymphocytic perivascular cuffing at the site of dermal injection was present in all turkeys given intradermal LPS.

Avian Dis, 1994 Oct-Dec, 38(4), 778 - 89
Partial purification of cross-protection factor(s) from Pasteurella multocida; Rimler RB; Membrane-associated cross-protection factor(s) (CPF) of in vivo-grown Pasteurella multocida were solubilized by detergent and partially purified by sequential chromatography on ion exchange, gel filtration, and hydroxyapatite columns . The CPF activity was determined by challenge of turkeys vaccinated with different chromatography fractions and challenge of poults passively immunized with serum from the same vaccinated turkeys . Analyses of different chromatography fractions by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblots suggested that proteins in the range of 59-65 kDa and a protein of 39-kDa molecular mass were associated with cross-protection . Although CPF activity was found to be associated with protein-containing material, treatment with periodate diminished cross-protection, indicating that carbohydrate determinants may also contribute to immune cross-protection.

Zentralbl Veterinarmed B, 1994 Oct, 41(7-8), 483 - 91
The role of eicosanoids in the chemotactic response to Pasteurella haemolytica infection; Clarke CR et al.; The chemotactic role of eicosanoids in the pathogenesis of Pasteurella haemolytica infection was studied, using a tissue chamber infection model and pharmacological inhibitors of eicosanoid synthesis . Tissue chambers were implanted subcutaneously in 12 calves allotted to three treatment groups of equal size . At 45 days after implantation, calves received saline, dexamethasone, or phenylbutazone treatments, and tissue chambers in all animals were then inoculated with P . haemolytica . Chamber fluid samples were collected before inoculation and at 2, 6, 18, 40, and 90 h after inoculation . Bacterial counts, total leukocyte counts, pH and albumin concentrations in chamber fluids were determined using standard bacteriological and clinical pathological methods . Concentrations of eicosanoids and activity of interleukin-1 (IL-1) were measured by radioimmunoassay and a helper T cell bioassay, respectively . Concentrations of leukotriene B4 (LTB4), thromboxane B2 (TXB2), 6-keto-prostaglandin F1 alpha (PGF1 alpha) and prostaglandin E2 (PGE2) increased markedly after inoculation . An inhibitory effect of dexamethasone on both LTB4 production and neutrophil influx, together with the temporal relationship between these two events, suggested that LTB4 served as a chemo-attractant . Activity-time profiles for IL-1 in chamber fluids were similar to those of the eicosanoids . Phenylbutazone and dexamethasone reduced the severity of the inflammatory responses as measured by lower concentrations of albumin and higher pH in treated versus control chamber fluids . The results of this study suggest that eicosanoid inflammatory mediators play an important chemotactic role in the pathogenesis of P . haemolytica infection.

J Vet Diagn Invest, 1994 Oct, 6(4), 442 - 7
Molecular fingerprinting of Pasteurella multocida associated with progressive atrophic rhinitis in swine herds; Gardner IA et al.; Ninety-six nasal isolates of Pasteurella multocida from swine herds with progressive atrophic rhinitis were characterized by restriction endonuclease analysis (REA) of whole-cell DNA, ribotyping, and plasmid analysis . For REA, bacterial DNA was digested with SmaI and electrophoresed in 0.7% agarose, and fragments were visualized with UV light . For ribotyping, EcoRI-digested and electrophoresed restriction fragments of whole-cell DNA were transferred to nitrocellulose membranes, hybridized with gamma-32P-labeled Escherichia coli ribosomal RNA, and visualized by autoradiography . Phenotypes of isolates were toxigenic capsular type D (n = 51), nontoxigenic type D (n = 28), nontoxigenic type A (n = 16), and toxigenic type A (n = 1) . Plasmids of various sizes were evident in 92.2% and 17.9% of toxigenic and nontoxigenic D strains, respectively, but were absent from all type A strains . Among the 4 phenotypes, there were 17 REA profiles and 6 ribotypes . For 3 of 17 REA patterns, multiple ribotypes were evident, and several REA types were evident in 5 of 6 ribotypes . Thirty-seven isolates of toxigenic capsular type D from Australian herds were either SmaI type B or C and ribotype 2, whereas 14 toxigenic D isolates from the USA and other countries were more heterogeneous (7 REA types and 6 ribotypes) . The fingerprinting results provided evidence in support of the hypothesis of a single source infection in Australia associated with the introduction of breeding pigs from overseas.

Infect Immun, 1994 Sep, 62(9), 3817 - 28
Characterization of the dermonecrotic toxin in members of the genus Bordetella; Walker KE et al.; All members of the genus Bordetella and Pasteurella multocida (a gram-negative bacillus genetically unrelated to Bordetella spp., yet often sharing the same ecological niche) produce a dermonecrotic toxin (DNT) . The amount of toxin produced and the time required for appearance of the lesions are identical for Bordetella pertussis, B . parapertussis, and B . bronchiseptica but different for P . multocida and B . avium . DNT has been reported to act by promoting vasoconstriction; however, vasoactive compounds (verapamil, prazosin, hydralazine, tolazoline, or isoxsuprine) are able to reverse the action of the toxin only slightly . Vasoconstrictors (atropine, serotonin, epinephrine, or endothelin) did not produce DNT-like lesions . We have characterized a region of DNA essential for DNT expression . We have determined by Southern analysis that the restriction map of the DNT gene is nearly identical in B . pertussis, B . parapertussis, and B . bronchiseptica, but the sequences are not present in toxigenic B . avium and P . multocida strains . A gentamicin resistance-origin of transfer cassette cloned into a 1.8-kb NotI-BamHI fragment results in constructs which can be mobilized and recombined into the Bordetella chromosome, rendering the resultant B . pertussis, B . parapertussis, and B . bronchiseptica strains negative for DNT . A 5-kb BamHI-ApaI fragment from the B . pertussis chromosome was sequenced and revealed homology to the Escherichia coli CNF1 (cytotoxic necrotizing factor 1) toxin.

Rev Sci Tech, 1994 Sep, 13(3), 837 - 43
Evaluation of three oil-adjuvant vaccines against Pasteurella multocida in buffalo calves; Muneer R et al.; Three oil-adjuvant vaccines of Pasteurella multocida 6:B were evaluated with respect to the level and duration of the humoral immune response produced in buffalo calves . Preparation 1 was a water-in-oil emulsion containing Marcol 52, Montanide 888 and antigen at a ratio of 6:1:3 . Preparation 2 was a double emulsion containing Marcol 52, Arlacel A and Tween 80 in addition to antigen . Preparation 3 contained alpha-d-tocopheryl acetate (vitamin E), Montanide 888 and antigen . All three preparations induced a similar sustained immune response in buffalo calves beyond 270 days post-vaccination.

Vet Microbiol, 1994 Sep, 42(1), 35 - 44
Construction and vaccine potential of an aroA mutant of Pasteurella haemolytica; Homchampa P et al.; The aroA gene, encoding 5-enolpyruvylshikimate 3-phosphate synthase, from Pasteurella haemolytica biotype A, serotype 1 was cloned by complementation of the aroA mutation in Escherichia coli strain AB2829 after electroporation with a DNA library constructed in pUC18 . The cloned P . haemolytica aroA gene was inactivated by insertion of a kanamycin resistance gene and reintroduced by allelic exchange into the chromosome of the parental P . haemolytica using PbluescriptII SK+ . The P . haemolytica aroA mutant was highly attenuated in a mouse septicaemic model . Mice immunized intraperitoneally with two doses of live P . haemolytica aroA mutant were protected against a lethal parental strain challenge.

Am J Vet Res, 1994 Sep, 55(9), 1267 - 74
Enhancement of Pasteurella haemolytica leukotoxic activity by bovine serum albumin; Waurzyniak BJ et al.; Growth of Pasteurella haemolytica A1 in RPMI 1640 medium containing 0.5% bovine serum albumin (BSA) for 2.5 hours enhanced culture supernatant leukotoxic activity (30,700 +/- 12,900 toxic units/ml, compared with leukotoxic activity of culture supernatants produced in RPMI 1640 medium alone (120 +/- 40 toxic units/ml) . Gel filtration chromatography of the leukotoxic activity from RPMI 1640 medium supernatants in buffer containing 50 mM NaCl indicated a single leukotoxic activity peak (peak I) eluting near the gel resin molecular mass exclusion limit (estimated molecular mass of approx 8,000 kd) . In contrast, culture supernatants produced in RPMI 1640 plus bovine serum albumin medium (RPMI + BSA) had peak I and 2 additional leukotoxic activity peaks (peaks II and III) with estimated molecular mass of approximately 80 and < 30 kd, respectively . All leukotoxic activity peaks were composed of approximately 100-kd molecular mass leukotoxin protomer, as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting with a monoclonal antibody against leukotoxin . Subjecting culture supernatant leukotoxic activity produced in RPMI + BSA to gel filtration chromatography in buffer containing 500 mM NaCl or 6M urea resulted in detection of only a single leukotoxic activity peak with estimated approximate molecular mass of 250 and 800 kd, respectively . These findings suggest that P haemolytica exists as a high molecular mass aggregate with low leukotoxic activity which, in the presence of BSA, partially disaggregates to multiple toxin forms with enhanced leukotoxic activity . Some of these leukotoxin forms interact with dextran-based gel resins at low ionic strength.

Microb Pathog, 1994 Sep, 17(3), 149 - 58
Expression, purification and immunologic analysis of three Pasteurella haemolytica A1 28-30 kDa lipoproteins; Dabo SM et al.; Three genes, tandemly arranged on the Pasteurella haemolytica A1 chromosome and encoding similar 28-30 kDa proteins, were previously cloned and sequenced by our laboratory . In this study, we demonstrate that the cloned genes encode lipoproteins, as previously suggested by DNA sequence analysis . To further analyze the bovine immune response to these proteins, the individual genes were cloned separately into an expression vector and recombinant forms of the three proteins were purified after expression in Escherichia coli . Sera from cattle vaccinated with live P . haemolytica or P . haemolytica bacterins and from cattle naturally exposed to P . haemolytica recognized each of the recombinant proteins . Vaccination with live or killed whole bacteria did not elicit an immune response of the same quality as that which developed in response to natural infection . A statistically significant correlation existed between resistance to challenge and a high antibody response to one of these three proteins.

FEMS Microbiol Lett, 1994 Aug 15, 121(2), 199 - 205
The detection of the sialoglycoprotease gene and assay for sialoglycoprotease activity among isolates of Pasteurella haemolytica A1 strains, serotypes A13, A14, T15 and A16; Lee CW et al.; Polymerase chain reaction (PCR) using specific primers to the sialoglycoprotease gene (gcp) of Pasteurella haemolytica biotype A, serotype 1 amplified a 1-kb fragment from each of P . haemolytica serotypes A7, A13, A14 and A16, but not T15; which was confirmed by Southern blot hybridization analysis . Using a sialoglycoprotease (Gcp) activity assay, Gcp activity was found in serotypes A13, A14 and A16 . Inclusion of these three serotypes confirms that all recognized A biotypes are positive for both gcp gene and activity, with the exception of serotype A11 (which has a different genetic organization and exhibits no Gcp activity) . Furthermore, all recognized T biotypes are negative for both the gene and Gcp activity.

Vet Microbiol, 1994 Aug 15, 41(4), 311 - 9
Bovine neutrophil activation by culture fluid from Pasteurella haemolytica serotypes A1 and A11; Mdurvwa EG et al.; Neutrophil activation has been thought to contribute to the pathogenesis of fibrinous pneumonia caused by Pasteurella haemolytica serotype A1 . The activation of bovine neutrophils by culture fluid from the pathogenic P . haemolytica serotype A1 and the non-pathogenic serotype A11 was compared . Logarithmic-phase bacteria of each serotype were incubated in RPMI 1640-medium for 3 h at 37 degrees C . The culture fluid was collected by centrifugation and concentrated by ultrafiltration . The concentrated culture fluids were then tested for their ability to induce chemotaxis and respiratory burst in bovine neutrophils . Chemotactic activity was of similar magnitude in response to both serotypes . An early chemiluminescence response occurred at 5 min at 1:100 dilution and a late peak at 11 min at 1:500 dilution for serotype A1 . The early peak was absent at all dilutions tested for serotype A11 . Maximal chemiluminescence response was observed at 1:25 dilution with serotype A11 while maximal response was seen at 1:500 dilution with the culture fluid from serotype A1 . Superoxide anion release was greater in response to culture fluid from A1 than A11 at all dilutions tested . Leukotoxin activity was 50-fold higher in culture fluid from serotype A1 than in culture fluid from serotype A11 . In this study, the ability of P . haemolytica to attract and activate bovine neutrophils was not restricted to the pathogenic serotype A1.

Appl Environ Microbiol, 1994 Aug, 60(8), 2990 - 8
Iron uptake by Pasteurella piscicida and its role in pathogenicity for fish; Magarinos B et al.; We evaluated the iron uptake mechanisms in Pasteurella piscicida strains as well as the effect of iron overload on the virulence of these strains for fish . With this aim, the capacity of the strains to obtain iron from transferrin and heme compounds as well as their ability to overcome the inhibitory activity of fish serum was analyzed . All the P . piscicida strains grew in the presence of the iron chelator ethylene-diamine-di (O-hydroxyphenyl acetic acid) or of human transferrin, which was used by a siderophore-mediated mechanism . The chemical tests and cross-feeding assays showed that P . piscicida produced a siderophore which was neither a phenolate nor a hydroxamate . Cross-feeding assays as well as preliminary chromatographic analysis suggest that this siderophore may be chemically related to multocidin . All the P . piscicida isolates utilized hemin and hemoglobin as an iron source, since the virulence of the strains increased when the fish were preinoculated with these compounds . This effect was stronger in the avirulent strains (50% lethal dose was reduced by 4 logs when fish were pretreated with hemin or hemoglobin) . Only the pathogenic P . piscicida isolates were resistant to the bactericidal action of the fresh fish serum . The nonpathogenic strains grew in fish serum only when it was heat-inactivated or when it was supplemented with ferric ammonium citrate, hemin, or hemoglobin . In all the strains, at least three iron-regulated outer membrane proteins (IROMPs) (105, 118, and 145 kDa) were increased when the strains were cultured in iron-restricted medium.(ABSTRACT TRUNCATED AT 250 WORDS)

Am J Vet Res, 1994 Aug, 55(8), 1107 - 10
Serotype-specific inhibition of colonization of the tonsils and nasopharynx of calves after Pasteurella haemolytica serotype A1 after vaccination with the organism; Frank GH et al.; Vaccination of cattle with a tissue culture-derived Pasteurella haemolytica serotype 1 vaccine elicited a serotype-specific inhibition of nasal and tonsillar colonization by the homologous serotype under field conditions . Calves (n = 101) originated from a single farm, where half the calves were vaccinated . The calves were delivered to an order-buyer barn 105 days later, and given a second dose of vaccine . At the order-buyer barn, calves were mixed with 27 calves, some of which had clinical signs consistent with respiratory tract disease . Also 12 of the original calves were infected with P haemolytica serotype 1 by tonsillar instillation . After 6 days at the order-buyer barn, calves were shipped 1,600 km by truck to a feedyard, and arrived the next day . Tonsillar wash and nasal secretion aspiration specimens were collected for culture of P haemolytica on days 1, 8, and 29 at the feedyard . Inhibition of colonization was evidenced by lower frequency of isolations from the vaccinates than from the nonvaccinates after transport to the feedyard . Selectively lowering the frequency of colonization by P haemolytica serotype 1 could reduce losses attributable to pneumonic pasteurellosis.

Vet Microbiol, 1994 Aug 1, 41(3), 221 - 33
Growth-condition dependent expression of Pasteurella haemolytica A1 outer membrane proteins, capsule, and leukotoxin; Gatewood DM et al.; Pasteurella haemolytica, strain P1148 (biotype A, serotype 1) was grown under iron-rich and iron-restricted conditions both with and without serum, and the outer membrane protein (OMP), capsule, and leukotoxin production studied . OMPs were evaluated by SDS-PAGE and examined by immunoblot to identify antigens recognized by sera from P . haemolytica A1 convalescent and vaccinated cattle . Capsule production was evaluated using fluorescent antibody staining and rapid plate agglutination reaction . Leukotoxin production was measured by neutrophil 51Cr-release assay . Expression of specific OMPs, amount and antigenic character of capsule, and quantity of leukotoxin produced by P . haemolytica A1 varied in response to alterations in the growth media . Immunoblots indicated the immune response of convalescent calves differs from vaccinated calves, and convalescent calves produce antibodies to novel OMPs induced by growth in iron-restricted conditions.

Poult Sci, 1994 Aug, 73(8), 1169 - 74
Genetic analysis of antibody responses of turkeys to Newcastle disease virus and Pasteurella multocida vaccines; Sacco RE et al.; Heritability (h2) of 16-wk BW and primary and secondary antibody responses and genetic and phenotypic correlations among these traits were estimated for 931 male and female turkeys vaccinated with Newcastle disease virus (NDV) and Pasteurella multocida . Turkeys from a line selected for 22 or 23 generations for increased 16-wk BW were vaccinated at 6 and 12 wk of age with blood samples collected 3 wk postvaccination . Antibody titers were determined using an ELISA method and transformed to log(e) for analysis . Heritability estimates for primary and secondary antibody responses to NDV were .380 +/- .070 (SE) and .296 +/- .063, respectively . For primary and secondary antibody responses to P . multocida, h2 estimates were .458 +/- .075 and .333 +/- .066, respectively . Heritability estimate for 16-wk BW was .404 +/- .071 . The genetic correlation between primary and secondary antibody responses to NDV was .491 +/- .150 . There was no genetic correlation between primary and secondary antibody responses to P . multocida . Although the genetic correlation between primary antibody responses to NDV and P . multocida was .292 +/- .159, the genetic correlation between secondary responses to the two antigens did not differ from zero . There were no genetic correlations between antibody responses and 16-wk BW . Similar results were observed for phenotypic correlations . Based on heritability and genetic correlation estimates, it would be possible to improve antibody responses to either NDV or P . multocida singularly; however, to improve antibody responses to both antigens, selection would have to be applied for each antigen.

Gene, 1994 Jul 22, 145(1), 81 - 5
Construction of conjugative shuttle and suicide vectors for Pasteurella haemolytica and P . multocida; Azad AK et al.; A shuttle cloning vector, pAKA16, and suicide derivatives pAKA19 and pAKA22 have been developed for gene transfer to Pasteurella haemolytica and P . multocida . pAKA16 was constructed by insertion of the lacZ alpha-peptide-encoding region and a multiple cloning site into a plasmid which was originally isolated from P . haemolytica serotype A1 . The vector encodes ampicillin resistance, and contains at least 14 unique restriction sites and the property of phenotypic identification of recombinant clones in Escherichia coli by insertional inactivation of beta-galactosidase activity . It can be transferred by conjugation to P . haemolytica or P . multocida and is stably maintained in both species . The type-II chloramphenicol acetyltransferase-encoding gene (cat), cloned into pAKA16, was stably expressed in both P . haemolytica and P . multocida . Plasmids pAKA19 and pAKA22 were constructed by replacement of the origin of DNA replication (ori) of pAKA16 with a ColE1-type ori from pBR322 or an ori of plasmid R6K (oriR6K) from pJM703.1, respectively . These derivatives replicate in E . coli, but not in either P . haemolytica or P . multocida, and are suitable for use as suicide vectors for these Pasteurella species.

Res Vet Sci, 1994 Jul, 57(1), 129 - 32
Immediate responses in serum TNF alpha and acute phase protein concentrations to infection with Pasteurella haemolytica A1 in calves; Horadagoda A et al.; The concentrations of tumour necrosis factor-alpha (TNF alpha), serum amyloid A (SAA) and haptoglobin were determined in serum samples taken from four calves in the 10 hours after their intra-tracheal inoculation with Pasteurella haemolytica serotype A1 . The concentration of haptoglobin did not increase but the concentration of SAA rose progressively from within two hours of inoculation . The concentration of TNF alpha reached a peak in all the animals two hours after inoculation but had returned to undetectable levels after a further four hours . TNF alpha is likely to be an important mediator of the acute phase response in cattle and SAA is a more rapid bovine acute phase protein than haptoglobin in its response to infection with P haemolytica.

J Vet Diagn Invest, 1994 Jul, 6(3), 326 - 34
Streptococcus suis infection in swine: a retrospective study of 256 cases . Part II . Clinical signs, gross and microscopic lesions, and coexisting microorganisms; Reams RY et al.; A retrospective study of 256 cases of naturally acquired Streptococcus suis infections in swine submitted to the Indiana Animal Disease Diagnostic Laboratory from 1985 to 1989 was undertaken to describe the clinical signs, lesions, and coexisting organisms associated with S . suis serotypes 1-8 and 1/2 . Infected pigs generally had clinical signs and gross lesions referable to either the respiratory system or to the central nervous system (CNS), but not both . Neurologic signs were inversely related to gross lesions in the respiratory tract (R2 = -0.19, P = 0.003), as were respiratory signs and gross lesions in the CNS (R2 = -0.19, P = 0.003) . Suppurative bronchopneumonia was the most common gross lesion observed (55.2%, overall) . Fibrinous and/or suppurative pleuritis, epicarditis, pericarditis, arthritis, peritonitis, and polyserositis were also reported . In 68% of the pigs, other bacteria in addition to S . suis were isolated . Escherichia coli (35.0%) and Pasteurella multocida (30.0%) were the most commonly recovered bacterial agents . Mycoplasma and viral agents were identified less often, and their role in the development of streptococcosis was difficult to assess . In pigs infected with serotypes 2-5, 7, 8, and 1/2, suppurative meningitis with suppurative or nonsuppurative encephalitis, suppurative bronchopneumonia, fibrinopurulent epicarditis, multifocal myocarditis, and cardiac vasculitis were the most common microscopic lesions observed, whereas pigs infected with serotype 1 generally presented with suppurative meningitis and interstitial pneumonia . Microscopic lesions were morphologically similar among serotypes and were also similar to those reported with other pyogenic bacteria.(ABSTRACT TRUNCATED AT 250 WORDS)

Avian Dis, 1994 Jul-Sep, 38(3), 626 - 9
Diseases and management of backyard chicken flocks in Chitungwiza, Zimbabwe; Kelly PJ et al.; To gather information on backyard chicken flocks in Chitungwiza, an urban center in Zimbabwe, 85 flock owners were interviewed . The mean flock size was 53 birds (range 1-650), and most birds were kept for meat, for either domestic consumption or local sale . Mean age at slaughter was 12.4 weeks (range 8-24) . None of the owners vaccinated their birds, and reported mortality rates were high (mean 25%), most commonly being associated with diseases causing eye and respiratory problems . Most owners complained of a lack of technical and veterinary advice . Commercial enzyme-linked immunosorbent assays on sera from 460 birds in 52 flocks showed that the birds had been exposed to avian reovirus (3%), avian leukosis virus (9%), avian encephalomyelitis virus (11%), Newcastle disease virus (27%), Mycoplasma gallisepticum and M . synoviae (33%), Pasteurella multocida (52%), infectious bursal disease virus (55%), reticuloendotheliosis virus (65%), and infectious bronchitis virus (86%) . Parasite infections were detected only rarely.

Avian Dis, 1994 Jul-Sep, 38(3), 621 - 5
A serosurvey using enzyme-linked immunosorbent assay for antibodies against poultry pathogens in ostriches (Struthio camelus) from Zimbabwe; Cadman HF et al.; Horseradish peroxidase-conjugated goat anti-ostrich IgG was raised and used in commercial enzyme-linked immunosorbent assay kits to detect antibodies reactive with 11 poultry pathogens in sera from 149 ostriches from nine farms around Zimbabwe . Antibodies were detected to turkey rhinotracheitis virus (99%), Newcastle disease virus (23%), avian reovirus (19%), infectious bursal disease virus (15%), avian encephalomyelitis virus (15%), Mycoplasma gallisepticum and/or M . synoviae (11%), reticuloendotheliosis virus (10%), Salmonella enteritidis (8%), avian leukosis virus (3%), infectious bronchitis virus (2%), and Pasteurella multocida (< 1%) . Although evidence of prior infection with turkey rhinotracheitis and newcastle disease virus was present on all farms tested, there was marked variation between farms in the prevalence of exposure to other poultry pathogens.

Avian Dis, 1994 Jul-Sep, 38(3), 506 - 14
Passive cross-protection provided by antisera directed against in-vivo-expressed antigens of Pasteurella multocida; Wang C et al.; Chicken antisera produced against the detergent-insoluble fraction of in vivo-grown P-1059 (serotype 3) Pasteurella multocida passively protected against heterologous X-73 (serotype 1) challenge . Likewise, antisera from chickens that survived infection by live P-1059 provided passive heterologous protection . The two antisera, which contained cross-protection antibodies, were adsorbed with the detergent-insoluble fraction of in vitro-grown P-1059 and still passively cross-protected against heterologous serotype challenge . The proteins of in vivo-grown P-1059 that had molecular weights of 204,000, 192,000, 179,000, and 153,000, which were recognized by the adsorbed antisera on immunoblots, may be responsible for cross-protection.

Vet Microbiol, 1994 Jul, 41(1-2), 29 - 40
Serotype and genus specific protein antigens of Pasteurella multocida, revealed by monoclonal antibodies; Ramdani et al.; Two monoclonal antibodies (mAbs) reacting with protein antigens of Pasteurella multocida strain M1404 were produced and designated DM-1 and DM-4 . DM-1 reacted only with the homologous Heddleston serotype 2 . On the other hand, DM-4 reacted with all the 16 Heddleston serotypes and with other Pasteurella spp., but not with any of the other genera tested . Both mAbs may thus be of significance for identification and classification . The antibodies could agglutinate P . multocida only if the cells were treated with 1N HCl . Neither of the mAbs could opsonise P . multocida for phagocytosis by mouse macrophages nor did they protect recipient mice against lethal infection . Active immunisation of mice gave rise to only partial protection against challenge infection, suggesting a minor role for these particular proteins in protection from infection.

Vet Rec, 1994 Jun 4, 134(23), 597 - 8
Antimicrobial susceptibility of Pasteurella haemolytica and Pasteurella multocida isolated from pneumonic ovine lungs; Diker KS et al.; The in vitro susceptibilities of Pasteurella haemolytica biotypes A and T and P multocida from pneumonic ovine lungs to penicillin, ampicillin, oxytetracycline, chloramphenicol, erythromycin, kanamycin, neomycin, gentamicin, streptomycin and lincomycin were determined by the disk diffusion method . All the isolates were sensitive to chloramphenicol and resistant to lincomycin . The isolates of P haemolytica biotype A were consistently more sensitive to penicillin, ampicillin, oxytetracycline, erythromycin and streptomycin than those of biotype T.

Berl Munch Tierarztl Wochenschr, 1994 Jun, 107(6), 181 - 5
{The typing of Pasteurella from cattle and swine}; van Truong D et al.; A total of 383 Pasteurella strains isolated from calves and pigs were typed by their phenotypic properties and allotted to the following species and subspecies: {table: see text} P . multocida ssp . septica, P . avium biotype 2, P . canis and P . canis biotype 2 could be isolated up till now only from calves, on the other hand P . multocida ssp . gallicida were found in pigs only.

Appl Environ Microbiol, 1994 Jun, 60(6), 2011 - 6
Molecular gene cloning and nucleotide sequencing and construction of an aroA mutant of Pasteurella haemolytica serotype A1; Tatum FM et al.; The aroA gene of Pasteurella haemolytica serotype A1 was cloned by complementation of the aroA mutation in Escherichia coli K-12 strain AB2829 . The nucleotide sequence of a 2.2-kb fragment encoding aroA predicted an open reading frame product 434 amino acids long that shows homology to other bacterial AroA proteins . Several strategies to inactivate aroA were unsuccessful . Gene replacement was finally achieved by constructing a replacement plasmid with aroA inactivated by insertion of a P . haemolytica ampicillin resistance fragment into a unique NdeI site in aroA . A hybrid plasmid was constructed by joining the aroA replacement plasmid with a 4.2-kb P . haemolytica plasmid which encodes streptomycin resistance . Following PhaI methylation, the replacement plasmid was introduced by electroporation into P . haemolytica NADC-D60, a plasmidless strain of serotype 1A . Allelic exchange between the replacement plasmid and the chromosome of P . haemolytica gave rise to an ampicillin-resistant mutant which grew on chemically defined P . haemolytica medium supplemented with aromatic amino acids but failed to grow on the same medium lacking tryptophan . Southern blot analysis confirmed that aroA of the mutant was inactivated and that the mutant was without a plasmid.

Appl Environ Microbiol, 1994 Jun, 60(6), 2006 - 10
Characterization of a restriction endonuclease, PhaI, from Pasteurella haemolytica serotype A1 and protection of heterologous DNA by a cloned PhaI methyltransferase gene; Briggs RE et al.; Pasteurella haemolytica is the leading cause of economic loss to the beef cattle industry in the United States and an important etiologic agent worldwide . Study of P . haemolytica is hindered by researchers' inability to genetically manipulate the organism . A new restriction endonuclease, PhaI, an isoschizomer of SfaNI (R . J . Roberts, Methods Enzymol . 65:19-36, 1980), was isolated from P . haemolytica serotype 1, strain NADC-D60, obtained from pneumonic bovine lung . PhaI recognizes the 5-base nonpalindromic sequences 5'-GCATC-3' and 5'-GATGC-3' . Cleavage occurs 5 bases 3' from the former recognition site and 9 bases 5' from the latter recognition site . A gene encoding a methyltransferase which protects against PhaI cleavage was cloned from P . haemolytica NADC-D60 into Escherichia coli . Whereas unmethylated plasmid DNA containing a P . haemolytica origin of replication was unable to transform P . haemolytica when introduced by electroporation, the same plasmid DNA obtained from E . coli which contained a cloned PhaI methyltransferase gene could do so . The data indicate that PhaI is an effective barrier to the introduction and establishment of exogenous DNA in P . haemolytica.

Vet Immunol Immunopathol, 1994 Jun, 41(3-4), 307 - 21
Immune responses of piglets to Pasteurella multocida toxin and toxoid; van Diemen PM et al.; Experimental atrophic rhinitis (AR), serum antibody titres and in vitro lymphoproliferation to Pasteurella multocida derived toxin (Pm-T) were studied in piglets . Specific immune responses to Pm-T and Pm-T induced conchae atrophy were compared with AR immunity . This immunity was initiated by the Nobi-VAC AR-T vaccine administered at various times with respect to Pm-T challenge . Animals challenged with Pm-T developed conchae atrophy, but no antibodies nor cellular immune responses to Pm-T were detected . Vaccination 3 weeks before Pm-T challenge protected pigs against breakdown of nasal bony tissues . This protection was accompanied by an increase of serum antibodies and in vitro lymphoproliferation to Pm-T . Animals vaccinated 10 days before or after Pm-T challenge also had antibodies and cellular immune responses . However, these animals developed AR . In vitro, Pm-T appeared mitogenic for quiescent (non-immune) peripheral lymphocytes and Concanavalin A stimulated lymphocytes from some pigs . These in vitro lymphoproliferative responses could be partly abrogated by the addition of monomorphic anti-swine major histocompatibility complex class II DQ and DR specific monoclonal antibodies . We conclude that Pm-T is poorly immunogenic in vivo and does not initiate a protective Pm-T specific immune response . Pigs were protected from AR by vaccination, but protection was dependent on the timing of vaccine administration . We speculate that Pm-T modifies the immune response such that the response is not directed towards the toxin but to an unidentified component in the nose of piglets.

Vet Microbiol, 1994 Jun, 40(3-4), 283 - 91
Identification of a potentially important antigen of Pasteurella haemolytica; Weldon SK et al.; To identify antigens which may be important for stimulating immunity to pneumonic pasteurellosis, a bovine antiserum to whole P . haemolytica was used to screen a recombinant lambda gt11/P . haemolytica expression library . One of the recombinant bacteriophage clones identified with the bovine antiserum, SW20C, expressed a fusion protein which was also recognized by rabbit antiserum to partially purified P . haemolytica culture supernatant and was found to be immunogenic in guinea pigs . The guinea pig antibody recognized a 100 kDa protein in P . haemolytica cell lysates . Sequence analysis of the cloned DNA from SW20C identified a fragment of 1446 bp with a small open reading frame that was contiguous with the lacZ sequence . The 153 bp P . haemolytica-specific open reading frame encoded a polypeptide of approximately 6 kDa . Homology searches of Genbank and the EMBL data bases revealed no homology of this open reading frame with any other bacterial sequences including P . haemolytica leukotoxin and Ssa1 . Evaluation of sera from calves that were scored either susceptible or resistant to experimental pneumonic pasteurellosis demonstrated a significant (P < 0.001) correlation between the intensity of the antibody response to the SW20C antigen and resistance to disease.

An Med Interna, 1994 Jun, 11(6), 297 - 8
{Spontaneous primary peritonitis due to Pasteurella multocida}; Navarro V et al.; We present a new case of bacterial spontaneous peritonitis cirrhosis, associated to an extremely rare germ as it is Pasteurella multocida . We describe its clinical characteristics and evolution after treatment.

Microb Pathog, 1994 Jun, 16(6), 423 - 33
Development of an intraperitoneal implant chamber for the study of in vivo-grown Pasteurella haemolytica in cattle; Davies RL et al.; An intraperitoneal implant chamber was developed for the study of the in vivo growth of Pasteurella haemolytica in calves . The chamber had a volume of approximately 100 ml and featured an external sampling port which allowed multiple and sequential sampling of the chamber contents . A single polycarbonate diffusion membrane with a pore size of 0.22 micron allowed host peritoneal fluid to diffuse into the chamber and maintained the bacterial population free of white blood cells . Chambers were implanted into the peritoneal cavities of four five-month-old dairy-cross calves, demonstrated to be sero-negative by indirect haemagglutination assay . Three days later, four different P . haemolytica isolates, of serotypes A1 or A2, were inoculated into the chambers . In all cases, there was a slow decline in the viable bacterial numbers within the chambers . Western blot analysis of the antibody content of the chamber fluids revealed IgG antibodies to P . haemolytica OMPs in the fluid prior to inoculation and both 9 and 15 days after inoculation . Furthermore, there was no significant change in the IgG antibody content of the chamber fluid, either quantitatively or qualitatively, during the course of the experiment . Analysis of the bactericidal activity of pre-inoculation chamber fluid against the corresponding bacterial isolate suggested that an antibody-dependent complement-mediated process was not responsible for the decline in bacterial numbers . Overall, the chamber design was demonstrated to be extremely effective for in vivo studies of P . haemolytica in calves, allowing easy and regular sampling of the chamber contents and maintaining bacteria free of white blood cells . Although there was a slow decline in bacterial numbers over time, sufficient numbers of cells could be obtained for analysis of cell-surface antigens.

Zentralbl Bakteriol, 1994 Jun, 281(1), 45 - 54
Isolation and partial characterization of the major protein of the outer membrane of Pasteurella haemolytica and Pasteurella multocida; Lubke A et al.; The N-terminal amino acid sequence of the 35 kDa (p35) major outer membrane protein (MOMP) of P . multocida shared a strong homology with those of homotrimeric nonspecific porins of gram-negative bacteria . The capacity of outer membrane protein (OMP) preparations of P . multocida to bind to respiratory mucosal surface preparations was inhibited significantly by using a polyclonal anti-p35 antiserum in an adhesion ELISA . Anti-p35 antiserum cross-reacted with a 44 kDa (p44) MOMP of P . haemolytica . N-terminal sequencing of MOMP p44 revealed a homology of 81% with the putative porin MOMP p35 of P . multocida.

Arch Biochem Biophys, 1994 May 1, 310(2), 300 - 9
Cleavage of epitectin, a mucin-type sialoglycoprotein, from the surface of human laryngeal carcinoma cells by a glycoprotease from Pasteurella haemolytica; Hu RH et al.; In this study we have assessed the action of a novel glycoprotease, secreted by the bovine pneumonia pathogen Pasteurella haemolytica, on epitectin expressed on the surface of human laryngeal carcinoma (H.Ep.2) cells . Epitectin has been previously characterized as a high buoyant density glycoprotein of mass of over 350 kDa extensively glycosylated on serine and threonine by small oligosaccharides . Purified metabolically labeled epitectin was very effectively hydrolyzed by the glycoprotease . However, short- and long-term treatments yielded a complex mixture of products which could not be resolved by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) or column chromatography, probably because of the heterogeneity of the structure and the distribution of the saccharides . Treatment of H.Ep.2 cells with glycoprotease followed by flow cytometric analysis revealed a significant loss in the cell surface epitopes detected by the anti-epitectin Ca2 monoclonal antibody . The action of the glycoprotease on cell surface epitectin was blocked by anti-glycoprotease antisera and was absent in an extract of a glycoprotease-negative strain of P . haemolytica . When extracts of cells treated with glycoprotease for 4 h were subjected to SDS-PAGE followed by 125I-wheat germ agglutinin overlay and autoradiography, the intensity of the characteristic epitectin bands was found to be drastically reduced compared to controls . H.Ep.2 cells metabolically labeled with {3H}glucosamine were also incubated with or without the glycoprotease and the released products were fractionated and analyzed . The enzyme-released products were found to be enriched in mucin-type glycopeptides . Thus the P . haemolytica glycoprotease could be used to selectively degrade mucin glycoproteins on cancer cell surface.

Infect Immun, 1994 May, 62(5), 1609 - 15
Regulation of major histocompatibility complex class II expression by Pasteurella haemolytica leukotoxin; Hughes HP et al.; Many properties have been associated with Pasteurella haemolytica leukotoxin and other repeat-in-toxin toxins, including their cytotoxic activity on various cells of the lymphoid and nonlymphoid systems as well as their ability to modulate the immunological activity of lymphocytes and monocytes . In this study, we assessed the ability of P . haemolytica leukotoxin to affect the expression major histocompatibility complex (MHC) class II molecules on bovine peripheral monocytes . Peripheral blood mononuclear cells were isolated from P . haemolytica leukotoxin-seronegative calves and incubated with various concentrations of authentic leukotoxin as well as the recombinant lktA gene product (LktA) . Expression of MHC class II antigen on cells was evaluated by flow cytometric methods . The results indicated that both a crude, authentic leukotoxin preparation and LktA were able to affect MHC class II expression by inducing a marked downregulation of MHC class II expression on bovine monocytes . However, when cells were activated with gamma interferon (IFN-gamma), LktA and Lkt had little or no detectable effect . By using a cell line which expresses MHC class II only after activation by INF-gamma, we were able to confirm the observation that LktA had no effect on the expression of MHC class II after IFN-gamma treatment . Leukotoxin affected the functional capacity of monocytes to present antigen, as demonstrated by the ability of LktA or authentic leukotoxin to totally inhibit a mixed lymphocyte culture from MHC-mismatched calves . Thus, leukotoxin was able to downregulate constitutive expression of MHC class II expression, and we propose that this is a novel way in which this molecule can affect the immune function of monocytes, playing an important role in bacterial pathogenesis and survival of organisms at the infection site.

Vet Immunol Immunopathol, 1994 May, 41(1-2), 89 - 100
Use of a 35.5 kDa cell membrane composition of Pasteurella multocida and an anti-idiotype antibody to induce protective immunity in leghorn chickens; Zhang H et al.; Using polyacrylamide gel electrophoresis and Western blot analysis, a 35.5 kDa cell membrane composition of Pasteurella multocida ATCC 11039 was identified as a dominant epitope of the bacterium . Mice inoculated with inactivated whole bacteria produce antisera primary reactive with the 35.5 kDa component (Pm35.5) in Western blot analysis . The outer membrane component was composed primarily of protein but did have lipopolysaccharide present at 133 micrograms mg-1 protein . In challenge trials (Trial 2), groups of white leghorn chickens vaccinated with Pm35.5 or an anti-idiotypic antibody (using Pm35.5 as the original antigen) had significant difference in mortality of 3.2% and 48.3%, respectively, when compared with unvaccinated controls (99%) . Mortality in a group of chickens receiving a commercially available bacterin (9.2%) was higher but not significantly different from the mortality of the Pm35.5 vaccinated group (3.2%).

Vet Immunol Immunopathol, 1994 May, 41(1-2), 73 - 88
Investigation of the poultry idiotypic network using Pasteurella multocida; Zhang H et al.; The idiotypic network in leghorn laying hens was investigated by inoculating hens with a 35.5 kD outer membrane protein of Pasteurella multocida (Pm35.5), idiotype (Id), or Pm35.5 anti-Id . Egg yolks were analyzed for the presence of Id, anti-Id, or anti-anti-Id . Anti-Pm35.5 antibodies (Id) were not titered to an end-point but were present in high concentrations . The presence of anti-Id antibody in yolk was demonstrated by the inhibition of Pm35.5 binding to Id by anti-Id using a flow microsphere inhibition immunoassay . Inhibition of Pm35.5 binding to Id caused by different anti-Id preparations ranges from 51.5 to 56.1% . Not all of the anti-Id bound to a paratope-associated Id, since 8.3-12.8% of the fluoresceinated anti-Id bound to Id-coated beads in the presence of excess Pm35.5 . We confirmed that a portion of the anti-Id antibodies was an internal image of the Pm35.5 Id and could mimic antigen by demonstrating that anti-Id inoculated in naive hens caused the synthesis of anti-anti-Id antibodies that bound to Pm35.5 in enzyme linked immunosorbent assay . Monoclonal anti-Id antibodies were also capable of inhibiting Pm35.5 binding to Pm35.5 Id coated microspheres and inducing anti-anti-Id antibodies in BALB/cJ mice that reacted with the original antigen, Pm35.5 . Our investigation has also shown that idiotypic antibodies to P . multocida were transferred from chicken to egg.

Vet Pathol, 1994 May, 31(3), 340 - 8
Distribution of anti-CD68 (EBM11) immunoreactivity in formalin-fixed, paraffin-embedded bovine tissues; Ackermann MR et al.; A commercially acquired anti-human macrophage antibody (anti-CD68; EBM11) was used in an immunocytochemical technique to detect macrophages in formalin-fixed, paraffin-embedded tissues from cattle, pigs, humans, rats, turkeys, dogs, and cats . In healthy cattle, the antibody labeled alveolar macrophages, pulmonary intravascular cells (presumably intravascular macrophages), and macrophage-like cells in other tissues . In bovine lungs infected with Pasteurella haemolytica, EBM11 antibody labeled 95% of alveolar macrophages and macrophages within alveolar septa but only 0-2% of streaming or "oat" leukocytes . Alveolar macrophages were also stained by EBM11 in pigs but not in rats, turkeys, dogs, and cats . The antibody also stained macrophage aggregates in the mesenteric lymph nodes and intestinal lamina propria of Mycobacterium paratuberculosis-infected cattle . This study shows that the anti-CD68 (EBM11) antibody is a useful marker of macrophages in normal bovine tissues or tissues from areas of acute or chronic inflammation that have been routinely processed . The study also adds strength to the growing evidence suggesting that streaming leukocytes seen in pneumonic pasteurellosis are neutrophils.

Proc Natl Acad Sci U S A, 1994 Apr 26, 91(9), 3814 - 8
Cytotoxic necrotizing factor type 2 produced by virulent Escherichia coli modifies the small GTP-binding proteins Rho involved in assembly of actin stress fibers; Oswald E et al.; Cytotoxic necrotizing factor type 2 (CNF2) produced by Escherichia coli strains isolated from intestinal and extraintestinal infections is a dermonecrotic toxin of 110 kDa . We cloned the CNF2 gene from a large plasmid carried by an Escherichia coli strain isolated from a lamb with septicemia . Hydropathy analysis of the deduced amino acid sequence revealed a largely hydrophilic protein with two potential hydrophobic transmembrane domains . The N-terminal half of CNF2 showed striking homology (27% identity and 80% conserved residues) to the N-terminal portion of Pasteurella multocida toxin . Methylamine protection experiments and immunofluorescence studies suggested that CNF2 enters the cytosol of the target cell through an acidic compartment and induces the reorganization of actin into stress fibers . Since the formation of stress fibers in eukaryotic cells involves Rho proteins, we radiolabeled these small GTP-binding proteins from CNF2-treated and control cells with a Rho-specific ADP-ribosyltransferase . The {32P}ADP-ribosylated Rho proteins from CNF2-treated cells migrated slightly more slowly in SDS/PAGE than did the labeled proteins from the control cells . This shift in mobility of Rho proteins in SDS/PAGE was also observed when CNF2 and the RhoA protein were coexpressed in E . coli . We propose that Rho proteins are the targets of CNF2 in mammalian cells.

Microbiol Res, 1994 Apr, 149(1), 89 - 93
{The influence of cations on the letality and on the formation of the toxin of Pasteurella multocida}; Erler W et al.; The stability of the Pasteurella multocida toxin has proven as very pH dependent . Therefore, a pH lower than 6 should be avoided . A concentration of 1 mM Cu2+ decreased the lethal effect of the toxin but the equimolecular addition of chelators compensated this effect . Possible structure-effect mechanisms were discussed . Iron supplement (100 microM) to two culture media examined did not influence the production of the toxin but during the cultivation in the low-iron medium TDHM the toxin level increased . This increased toxin production was not due to a more intensive multiplication of bacteria.

Lab Anim, 1994 Apr, 28(2), 130 - 7
Colonization and antibody response in mice and rats experimentally infected with Pasteurellaceae from different rodent species; Boot R et al.; Mice and rats were experimentally infected with Pasteurellaceae isolated from mice, rats, hamsters and gerbils . Mice and rats were most heavily colonized by strains originally isolated from mice and rats respectively, and to a lesser extent by Pasteurellaceae from hamsters and gerbils . Colonization was generally accompanied by seroconversion . Gross pathology of the lungs was not observed . We conclude that Pasteurellaceae-free SPF mice and rats can be colonized by members of this bacterial family present in other rodent species.

J Wildl Dis, 1994 Apr, 30(2), 263 - 6
A reliable transport method for isolating Pasteurella haemolytica from bighorn sheep; Foreyt WJ et al.; We compared three transport methods for the recovery of Pasteurella haemolytica from pharyngeal swabs collected under field conditions from 42 bighorn sheep (Ovis canadensis) in one captive and three free-ranging populations . Transport methods included: Amies medium with charcoal, transported on ice, and cultured on blood agar within 24 hr; direct plating on blood agar, transported on heating pads, and incubated at 37 C within 8 hr of collection; and phosphate buffered glycerol (PBG), transported on dry ice, and stored at -70 C for 10 days before culture . Isolates of P . haemolytica were recovered from all 42 bighorn sheep with a mean (+/- SE) of 1.2 +/- 0.1 (Amies), 1.3 +/- 0.1 (blood agar), and 1.3 +/- 0.1 (PBG) isolates per swab . No statistical differences (P > 0.05) were observed in the recovery of P . haemolytica isolates among the transport methods . However, based on our experience and results of this study, we recommend that if submission of samples to the laboratory is likely to be delayed, pharyngeal swabs be transported in PBG on dry ice and kept frozen until they are cultured . Viable samples can be maintained in PBG at -70 C for several years.

J Wildl Dis, 1994 Apr, 30(2), 137 - 45
Fatal pneumonia following inoculation of healthy bighorn sheep with Pasteurella haemolytica from healthy domestic sheep; Foreyt WJ et al.; In a series of three experiments, isolates of Pasteurella haemolytica biotype A, serotype 2, ribotype reference WSU-1, from healthy domestic sheep, were inoculated intratracheally into eight bighorn sheep (Ovis canadensis canadensis) and seven domestic sheep with doses of bacteria ranging from 5.3 x 10(8) to 8.6 x 10(11) colony forming units . Seven of eight inoculated bighorn sheep died from acute pneumonia within 48 hr of inoculation, whereas all seven domestic sheep inoculated with comparable or greater doses of bacteria remained healthy . One contact control bighorn sheep also died 6 days after its penmates received P . haemolytica . Three other noncontact control bighorn sheep remained healthy during the experiments . Pasteurella haemolytica biotype A, serotype 2, ribotype reference WSU-1 in the inocula was recovered from one or more tissues from all bighorns that died; whereas, it was not detected in any bighorn sheep before inoculation . Three different ribotypes of P . haemolytica A2 were recovered from bighorn sheep; however, only the ribotype reference WSU-1 in the domestic sheep-origin inoculum was recovered from all dead bighorn sheep, and was not recovered from bighorn sheep that survived the experiments . Thus, a relatively nonpathogenic and common isolate of P . haemolytica from healthy domestic sheep was lethal in bighorn sheep under experimental conditions.

J Clin Microbiol, 1994 Apr, 32(4), 1004 - 10
Differentiation of toxigenic from nontoxigenic isolates of Pasteurella multocida by PCR; Nagai S et al.; A PCR assay was developed for the differentiation of toxigenic Pasteurella multocida subsp . multocida strains, the major etiologic agent for progressive atrophic rhinitis in pigs, from nontoxigenic strains . The PCR targeted a toxA gene encoding a 143-kDa dermonecrotic toxin that is considered to be the central etiologic factor in progressive atrophic rhinitis . toxA fragments were amplified from toxigenic P . multocida isolates but not from nontoxigenic isolates or other bacteria isolated from pigs . The sensitivity of the reaction was as low as 10 pg of chromosomal DNA from a toxigenic strain . The results obtained by PCR of the DNAs of 187 field isolates of P . multocida were consistent with those obtained by the guinea pig skin test and Western blot (immunoblot) analysis . Restriction fragment analysis of the PCR-amplified fragments from 67 field isolates and comparison of the DNA sequences of fragments from capsular serotype A and D strains suggest that the PCR-amplified region, which is considered to encode the major immunologic determinants of the toxin, would be the same among P . multocida strains . The PCR that we describe should be useful for the diagnosis and the etiologic survey of progressive atrophic rhinitis.

Am J Vet Res, 1994 Apr, 55(4), 502 - 9
Evaluation of an orally administered vaccine, using hydrogels containing bacterial exotoxins of Pasteurella haemolytica, in cattle; Bowersock TL et al.; Poly(methacrylic acid) hydrogels were tested for oral delivery of a vaccine against Pasteurella haemolytica infection in cattle . Culture supernatants of P haemolytica, the most common bacterium associated with pneumonia in cattle, were used as the antigens in the vaccine . Hydrogels containing culture supernatants were administered orally to calves . Calves were then challenge-exposed with virulent P haemolytica . Calves were euthanatized 3 days after challenge exposure . The lungs of each calf were scored for severity and size of pneumonic lesions . Results indicated that vaccinated calves had smaller, less severe pneumonic lesions and lived longer than nonvaccinated calves . These results indicated that hydrogels can be used to deliver vaccines orally to calves to enhance resistance to pneumonia caused by P haemolytica.

Microbiology, 1994 Apr, 140 ( Pt 4), 807 - 14
Variation in outer-membrane protein and lipopolysaccharide profiles of Pasteurella haemolytica isolates of serotypes A1 and A2 obtained from pneumonic and healthy cattle; McCluskey J et al.; The outer-membrane protein (OMP) and lipopolysaccharide (LPS) profiles of 29 isolates of Pasteurella haemolytica serotypes A1 (18 isolates) and A2 (11 isolates), obtained from pneumonic (13 isolates) or healthy (16 isolates) cattle, were compared by SDS-PAGE and Western blot analysis . Coomassie-blue-stained OMP profiles of serotype A1 and A2 isolates could be distinguished from each other by differences in both major and minor proteins . Whereas the OMP profiles of the serotype A1 isolates were extremely uniform in stained gels, there was variation in the mobilities of high-molecular-mass minor proteins and one of the major proteins of serotype A2 isolates . Differences in the OMP profiles of isolates within both the A1 and A2 serotypes were more clearly distinguished by Western blotting than by staining after SDS-PAGE . Thus, by Western blot analysis, four distinct OMP profiles were identified within the serotype A1 and A2 isolates, respectively . The profiles of the serotype A1 isolates were designated OMP types 1.1, 1.2, 1.3 and 1.4; those of the serotype A2 isolates were designated OMP types 2.1, 2.2, 2.3 and 2.4 . Three distinct LPS profiles were recognized among the isolates which, by comparison with previously described LPS types, were identified as smooth LPS type 1 and rough LPS types 3 and 5 . Isolates of serotype A1 consisted of LPS type 1 only . whereas isolates of serotype A2 consisted of LPS types 3 or 5 . OMP and LPS analysis of P . haemolytica has applications in epidemiological and virulence studies.

Can J Vet Res, 1994 Apr, 58(2), 93 - 8
Sialoglycoprotease of Pasteurella haemolytica A1: detection of antisialoglycoprotease antibodies in sera of calves; Lee CW et al.; Log phase culture supernate from Pasteurella haemolytica biotype A, serotype 1 contains a proteolytic enzyme specific for O-sialoglycoproteins . Using two methods, Western immunoblotting and enzyme neutralization assay, it was demonstrated that certain bovine sera from two previous P . haemolytica A1 vaccination and challenge trials contained antibodies (Ab) (isotypes IgG1 and IgG2 on Western immunoblot) to the sialoglycoprotease (Gcp) . In these trials, selected calves were vaccinated twice with either the commercial culture supernate vaccine Presponse or given phosphate-buffered saline (PBS) . One trial was conducted during spring, P . haem XIX, and the other during the winter, P . haem XXI . Although there was no clear evidence for induction of anti-Gcp in response to vaccination, several calves seroconverted following intrapulmonary challenge with live P . haemolytica A1 . This is the first report of anti-Gcp Ab in bovine sera . The results indicated that the Gcp is immunogenic and that the bacterium produces the enzyme in vivo . Further, animals with an anti-Gcp response had less pneumonia at necropsy, suggesting the Gcp may induce protective immunity.

Can J Vet Res, 1994 Apr, 58(2), 79 - 82
Induction of acute bronchopneumonia in mice by intrabronchial inoculation of Pasteurella haemolytica serotype 1; Pace LW et al.; Dose dependent pulmonary lesions of acute bronchopneumonia were induced in male, outbred Swiss Webster mice by intrabronchial inoculation of Pasteurella haemolytica . Five exponential dilutions ranging from 5 x 10(4) to 5 x 10(8) colony forming units per mL (CFU/mL) of Pasteurella haemolytica serotype 1 were inoculated into five groups of mice . Mice were killed by cervical dislocation 24 hours postinoculation . Pulmonary lesions occurred in mice of all five groups, however, 5 x 10(7) CFU/mL was the minimal dose which consistently produced lesions . Focal parenchymal necrosis, suppurative bronchiolitis, and flooding of interalveolar septa and alveoli by edema fluid, fibrin, neutrophils and macrophages, were observed microscopically . We conclude that outbred Swiss Webster mice can be used as a model for the study of selected disease mechanisms of acute lung inflammation and that this model may be used to determine some of the pathogenic mechanisms involved in the development of pulmonary lesions in bovine pneumonic pasteurellosis.

Avian Dis, 1994 Apr-Jun, 38(2), 390 - 2
Recurrent outbreaks of a cutaneous form of Pasteurella multocida infection in turkeys; Frame DD et al.; Cutaneous infection caused by Pasteurella multocida was diagnosed in a flock of seven thousand 17-to-22-week old male turkeys . The affected grow-out facility had an annual outbreak of fowl cholera, in which a cutaneous infection ventral and lateral to the tail was the predominant lesion . P . multocida serotypes 1 and 14 were the predominant isolates . The exact source of the infection was not determined.

Avian Dis, 1994 Apr-Jun, 38(2), 371 - 5
Serological survey of infections in waterfowl in the Guadalquivir marshes (Spain); Astorga RJ et al.; A serological survey was performed in the Guadalquivir Marshes (Southern Spain) in order to determine the presence and diffusion of six infective agents in wild waterfowl . The analysis covered 712 waterfowl from 13 species belonging to five taxa (Podicipediformes, Ciconiiformes, Anseriformes, Gruiformes, and Charadriiformes) . A range of immunological techniques led to detection of antibodies against the six infective agents studied: Chlamydia psittaci (13.3%), Mycoplasma anatis (3.5%), Pasteurella anatipestifer (2.9%), duck plague virus (1.6%), Aspergillus fumigatus (1.1%), and Newcastle disease virus (NDV) (0.2%) . The survey results indicated that these infective agents exist and circulate among wild waterfowl in the Guadalquivir Marshes.

Avian Dis, 1994 Apr-Jun, 38(2), 354 - 7
Effect of three commercially available adjuvants on the production of antibodies to Pasteurella multocida in broilers; McClimon LB et al.; Three commercially available adjuvants--incomplete Freund's, Ribi adjuvant system, and Hunter's TiterMax--were used to compare antibody responses of broilers to Pasteurella multocida antigen . Differences among treatments over the 9-week study were determined by measuring antibody responses of twice-vaccinated birds using an enzyme-linked immunoassay . The antibody level induced by the vaccine with incomplete Freund's adjuvant was the highest, followed by the vaccines with TiterMax and Ribi.

Avian Dis, 1994 Apr-Jun, 38(2), 334 - 40
Identification of common antigens of serotype 1 and serotype 3 Pasteurella multocida in poultry expressed in vivo; Wang C et al.; Eight groups of chicken immune sera and one group of turkey immune sera were used to identify unique in vivo-expressed antigens of Pasteurella multocida by Western immunoblotting . When antisera were prepared against the detergent-insoluble fraction of in vivo-grown P-1059, live X-73, live P-1059, live 86-1913, live P-1662, and live P-1702, they recognized four major antigens on immunoblots of the detergent-insoluble fraction of in vivo-grown serotype 1 and in vivo-grown serotype 3 . These four antigens, which have approximate molecular weights of 204,000, 192,000, 179,000, and 153,000, were not recognized by antisera made against the detergent-soluble fraction of in vivo-grown P-1059, whole in vitro-grown inactivated X-73, or whole in vitro-grown inactivated P-1059 . The results provide evidence for common antigens expressed by in vivo-grown serotype 1 and in vivo-grown serotype 3 . These antigens are recognized by sera from chickens that have survived infection by various serotypes . The antigens are concentrated in the detergent-insoluble fraction of in vivo-grown bacteria and likely represent unique in vivo-expressed outer-membrane proteins.

Avian Dis, 1994 Apr-Jun, 38(2), 317 - 24
A new selective enrichment procedure for isolating Pasteurella multocida from avian and environmental samples; Moore MK et al.; A selective enrichment procedure, using two new selective media, was developed to isolate Pasteurella multocida from wild birds and environmental samples . These media were developed by testing 15 selective agents with six isolates of P . multocida from wild avian origin and seven other bacteria representing genera frequently found in environmental and avian samples . The resulting media--Pasteurella multocida selective enrichment broth and Pasteurella multocida selective agar--consisted of a blood agar medium at pH 10 containing gentamicin, potassium tellurite, and amphotericin B . Media were tested to determine: 1) selectivity when attempting isolation from pond water and avian carcasses, 2) sensitivity for detection of low numbers of P . multocida from pure and mixed cultures, 3) host range specificity of the media, and 4) performance compared with standard blood agar . With the new selective enrichment procedure, P . multocida was isolated from inoculated (60 organisms/ml) pond water 84% of the time, whereas when standard blood agar was used, the recovery rate was 0%.

FEBS Lett, 1994 Mar 28, 342(1), 81 - 4
Mutation of a putative ADP-ribosylation motif in the Pasteurella multocida toxin does not affect mitogenic activity; Ward PN et al.; Pasteurella multocida toxin (PMT) is a potent mitogen for Swiss 3T3 fibroblasts and cytotoxic to embryonic bovine lung cells . Site-directed mutagenesis was used to investigate the functional significance of a three amino acid motif in PMT that is present in five other bacterial protein toxins which exhibit ADP-ribosyl transferase activity . Crude lysates of mutant clones were fully cytotoxic for embryonic bovine lung cells . Purified mutant toxin was also as effective at stimulating inositol phosphate turnover and nucleic acid synthesis as wild type toxin . We conclude that this motif has no functional significance in Pasteurella multocida toxin.

J Biol Chem, 1994 Mar 18, 269(11), 8260 - 7
Preferential inhibition of platelet-derived growth factor-stimulated DNA synthesis and protein tyrosine phosphorylation by nordihydroguaiaretic acid; Domin J et al.; Nordihydroguaiaretic acid (NDGA), a reportedly specific lipoxygenase inhibitor, was found to selectively inhibit platelet-derived growth factor (PDGF)-stimulated DNA synthesis in Swiss 3T3 cells . Maximal inhibition of PDGF-induced {3H}thymidine incorporation (96%) was observed using 4 microM NDGA (IC50 = 1.5 microM) . No effect of NDGA was observed upon DNA synthesis stimulated with either fetal bovine serum, bombesin, or epidermal growth factor (EGF) in the presence of insulin, or with the potent mitogen Pasteurella multocida toxin . The selective inhibition of PDGF-stimulated DNA synthesis by NDGA was also observed in diploid murine cells, rat, and human fibroblasts . Furthermore, 4 microM NDGA also inhibited PDGF-stimulated anchorage-independent colony growth of rat-1 cells by 76% . Using Swiss 3T3 cells, we found that PDGF-stimulated arachidonic acid mobilization and prostaglandin E2 production was abolished by NDGA in a dose-dependent manner . Inhibition of PDGF-stimulated arachidonic acid mobilization by NDGA could not, however, explain its potent inhibitory effect upon PDGF-stimulated DNA synthesis . Our results showed that NDGA also selectively inhibited PDGF receptor tyrosine phosphorylation in a dose-dependent manner in intact cells . Protein tyrosine phosphorylation stimulated by EGF or bombesin was not altered by NDGA treatment . Crucially, NDGA inhibited in vitro the tyrosine kinase activity of anti-phosphotyrosine and anti-PDGF receptor immunoprecipitates prepared from cultures stimulated with PDGF . This inhibition of receptor tyrosine phosphorylation in a cell-free system confirmed that NDGA acts directly at the level of the PDGF receptor tyrosine kinase domain . These results suggest that the potent and selective inhibitory effect of NDGA on PDGF-stimulated DNA synthesis results from its inhibitory action on tyrosine phosphorylation.

Vet Rec, 1994 Mar 12, 134(11), 263 - 6
The bovine placentome in bacterial and mycotic abortions; Johnson CT et al.; Placentomes were extracted from the uteri of 22 aborted cows and examined to detect the cause of abortion; fetuses or fetal abomasal contents from 15 of the cows were also examined . Firm diagnoses of Pasteurella haemolytica, Actinomyces pyogenes, Listeria monocytogenes, Bacillus licheniformis, Aspergillus fumigatus or Mortierella wolfii abortion were made in 11 cases . The histopathological lesions showed some correlation with the identity of the bacterium isolated; the lesions of mycotic abortion were distinct and characterised by a coagulative necrosis . The removal of a placentome was not followed by any observable deleterious effects.

Vet Microbiol, 1994 Mar, 39(1-2), 179 - 85
Physiological characteristics, antimicrobial susceptibility and serotypes of Pasteurella anatipestifer isolated from ducks in Thailand; Pathanasophon P et al.; Twenty isolates of Pasteurella (Moraxella) anatipestifer from ducks with serositis and septicemia in Thailand between 1988 and 1989 were characterized by various tests . Eighteen isolates fermented glucose and maltose, 3 fructose and 1 each mannose, arabinose, trehalose or sorbitol . All isolates produced gelatinase but not urease, while 2, 3, 5 and 6 produced indole, were CAMP positive, and were proteolytic for milk and coagulated serum respectively . Seven enzymes, phosphatase alkaline, esterase (C4), esterase lipase (C8), leucine arylamidase, valine arylamidase, phosphatase acid and phosphoamidase were detected from all the isolates . The isolates were highly susceptible to ampicillin, erythromycin, penicillin G and tylosin . Gel-diffusion precipitin tests demonstrated that serotype 1 was most prevalent (60%) and serotype 6 followed (5%) . Seven isolates (35%) were untypable . These results indicated that P . anatipestifer of serotype 1 played an important role in recent outbreaks of the disease in Thailand.

Am J Vet Res, 1994 Mar, 55(3), 348 - 52
Antigenic composition of Pasteurella haemolytica serotype-1 supernatants from supplemented and nonsupplemented media; Mosier DA et al.; Supernatant preparations containing antigens produced by Pasteurella haemolytica serotype 1 grown in nonsupplemented RPMI 1640 medium and 3 grown in supplemented RPMI 1640 media were evaluated . Antigens were detected by immunoblotting each supernatant preparation with sera from 20 cattle with various degrees of resistance to experimentally induced pneumonic pasteurellosis . Antigen-antibody bands at 49 and 30 kd were correlated with resistance in all 4 media . A 66-kd antigen-antibody band was correlated with resistance in 2 media, and antigen-antibody bands at 100 and 16 kd were correlated with resistance in 1 medium each . These results indicated that the number and relative amount of resistance-associated antigens in P haemolytica supernatants can be optimized on the basis of type of growth medium used.

Res Vet Sci, 1994 Mar, 56(2), 158 - 63
Effect of Pasteurella haemolytica infection on alpha 1-acid glycoprotein and albumin concentrations in serum and subcutaneous tissue chamber fluid of calves; Walker JL et al.; Thermoplastic tissue chambers were implanted subcutaneously in the paralumbar fossae of 10 calves . Concentrations of alpha 1-acid glycoprotein (AAG) and albumin in serum and subcutaneous tissue chamber fluid were measured before and after inoculation of Pasteurella haemolytica into tissue chambers . Two months after implantation, serum and tissue chamber fluid samples were collected and all tissue chambers were then inoculated with P haemolytica . Additional serum and chamber fluid samples were collected two, four, six and 10 days after inoculation . The concentrations of AAG and albumin in the samples were measured by radial immunodiffusion assay and the bromcresol method, respectively . P haemolytica inoculation resulted in an increase in the serum and chamber fluid AAG concentrations and an increase in chamber fluid albumin concentrations, suggesting that the proportion of drugs bound to serum and interstitial proteins may be affected by P haemolytica infection.

Kansenshogaku Zasshi, 1994 Mar, 68(3), 403 - 6
Brain abscess due to Pasteurella multocida; Li ZX et al.; The patient was a 26-year-old man who complained of headache and vomiting . On examination, there was nothing abnormal, but the edge of the right optic papilla was not clear . His temperature was 38.5 degrees C, pulse 96/min, blood pressure 120/80 mmHg . A space-occupying lesion in his fronto-dextra cupular part was found by CT scanning . He had a 12-year history of chronic purulent otitis . The diagnosis was a brain abscess in the fronto-dextra cupular part . The brain abscess was extracted and Pasteurella multocida was isolated from the dark brown pus draining from the abscess . The patient recovered through proper antibiotic therapy based on a sensitivity test . Reports of infections caused by this organism in foreign countries very widely from local infections due to bites and scratches by cats, dogs etc . to general infections such as infections of the respiratory tract, sepsis and meningitis . However, Pasteurella multocida brain abscesses are rare . Pasteurella multocida is a Gram-negative short rod which is best known as part of the mouth flora and as a pathogen causing septicemia in many domestic animals, such as cats, dogs etc. . Infection in man results mainly from animal bites or scratches . It has been reported that Pasteurella multodida can cause human septicemia, meningitis, respiratory tract infection, conjunctivitis and other infections . We isolated a strain of Pasteurella multocida from the pus of a brain abscess following chronic purulent otitis on August 6, 1990.

Infect Immun, 1994 Mar, 62(3), 1128 - 30
Pasteurella multocida produces heat shock proteins in turkeys; Love BC et al.; Pasteurella multocida produces an acute septicemic disease of turkeys . Since turkeys have a normal body temperature of 42 degrees C, it follows that P . multocida would produce heat shock proteins during the course of the infectious process . We show here that P . multocida produces several proteins at 42 degrees C, but not at 32 degrees C, and vice versa . Four of these proteins (70, 60, 40, and 35 kDa in molecular mass) were recognized by serum obtained from a turkey infected with P . multocida, suggesting that they were produced in vivo.

Clin Infect Dis, 1994 Mar, 18(3), 336 - 8
Infective endocarditis due to Pasteurella dagmatis: case report and review; Sorbello AF et al.; We report a case of endocarditis due to Pasteurella dagmatis complicated by vertebral osteomyelitis in a 55-year-old former alcoholic woman . To our knowledge, this is the second report of endocarditis attributed to that organism in the English-language medical literature . Rare cases of endocarditis due to other Pasteurella species and their varied clinical features are summarized.

FEMS Microbiol Lett, 1994 Feb 15, 116(2), 225 - 30
Preparation of recombinant glycoprotease of Pasteurella haemolytica A1 utilizing the Escherichia coli alpha-hemolysin secretion system; Lo RY et al.; Three murine monoclonal antibodies were prepared against the recombinant glycoprotease of Pasteurella haemolytica A1 expressed in Escherichia coli . These monoclonal antibodies were able to recognize the authentic glycoprotease from P . haemolytica A1 culture supernatant . A recombinant plasmid which contained most of the glycoprotease gene of P . haemolytica A1 fused with the secretion signal sequence from hlyA of the E . coli alpha-hemolysin determinant was constructed . This recombinant plasmid expressed a fusion protein (Gcp-F) which was secreted into the culture supernatant by E . coli cells when the alpha-hemolysin secretion functions HlyB and HlyD are supplied in trans . Gcp-F could be readily recovered from the supernatant free from other cellular materials and is suitable for use in vaccine trials and challenge experiments in animals.

Tijdschr Diergeneeskd, 1994 Feb 15, 119(4), 99 - 101
{Respiratory problems, growth retardation and arthritis in turkeys and broilers caused by a Pasteurella-like organism: Ornithobacterium rhinotracheale or 'Taxon 28'}; van Beek PN et al.; Since August 1993 moderate to serious respiratory problems with necrotic pneumonia, growth depression and fast increasing mortality are seen in commercial turkeys (2-8 weeks of age) and broilers (4-6 weeks of age) . An unidentified pleiomorphic Gram-negative rod was isolated from affected tissues . This Pasteurella-like organism, with yet unknown taxonomy, is recently named Ornithobacterium rhinotracheale gen . nov . sp . nov . or 'Taxon 28' . Experimentally severe growth depression and arthritis could be evoked in commercial turkeys and chickens . Respiratory signs caused by O . rhinotracheale could not (yet) be reproduced experimentally . This is the first report of the isolation of this organism in poultry in the Netherlands findings.

Microbiology, 1994 Feb, 140 ( Pt 2), 263 - 70
Modulation of Pasteurella multocida capsular polysaccharide during growth under iron-restricted conditions and in vivo; Jacques M et al.; Addition of the iron chelators 2,2'-dipyridyl, deferoxamine mesylate or apo-transferrin to culture media affected the composition and the morphology of Pasteurella multocida cells . Cells grown under iron-restricted conditions expressed iron-regulated proteins and, in addition, iron deprivation markedly reduced the amount of capsular material covering the cells of P . multocida . The addition of iron neutralized the effect of these chelators on capsule production . Cells of P . multocida grown under iron-restricted conditions were more labelled by gold particles coated with polymyxin which is known to interact with the lipid A-core region of lipopolysaccharides, and showed increased affinity for porcine respiratory tract mucus than cells grown under iron-sufficient conditions . Bacterial cells grown in vivo in peritoneal chambers in rats were also only covered by a thin layer (15-20 nm) of capsular material . Although the capsule is believed to be an important virulence factor, our data indicate that under iron-restricted conditions, such as those encountered in vivo, P . multocida may not be heavily encapsulated.

DNA Cell Biol, 1994 Feb, 13(2), 171 - 81
Static DNA bending and protein interactions within the Pasteurella haemolytica leukotoxin promoter region: development of an activation model for leukotoxin transcriptional control; Highlander SK et al.; In this study, we define cis-acting elements involved in regulation of the Pasteurella haemolytica leukotoxin promoter . In place of a canonical promoter -35 sequence, the leukotoxin promoter contains four adenine-rich repeats of sequence CA6(C/T)A, phased at approximately 10-base intervals . DNA fragments containing these repeats exhibit retarded mobility in polyacrylamide gels and permitted identification of a static DNA bend in the promoter -70 region . Deletion of the static DNA bend caused a two-fold reduction of leukotoxin transcription in Escherichia coli, suggesting that it is involved in promoter regulation . Three putative upstream activator sites (UAS), similar to those that bind the NifA activator in Klebsiella pneumoniae, are found 130 bp upstream from the transcription start site and are protected from DNase I cleavage by a P . haemolytica-specific factor . The promoter region also binds the DNA bending protein, the integration host factor (IHF), although IHF mutations do not affect its expression in E . coli . The arrangement of these elements suggests that leukotoxin expression is activated by a factor that interacts with the UAS and regulates transcription initiation at a distance via DNA looping . Activation and DNA bending may also influence a second, divergent promoter that lies 340 bp upstream from the leukotoxin start site.

Vet Microbiol, 1994 Feb, 38(4), 351 - 7
Phenotypic characterization of Zimbabwean isolates of Pasteurella multocida; Mohan K et al.; The phenotypic characteristics of 60 Zimbabwean isolates of Pasteurella multocida sensu stricto, from disease syndromes in different host species were studied . A number of representative strains were also serotyped . Consistent results were obtained in the tests for; catalase, oxidase, urease, indole, acid in glucose, inositol, salicin and sucrose . There was no obvious relationship between serotype, host or disease and the pattern of utilization of certain substrates by an isolate . This has been discussed in the context of recent proposals to reclassify Pasteurella and P . multocida on genotypic and phenotypic studies . It is suggested that notwithstanding the relevance of genetic studies in circumscribing P . multocida, the phenotype and disease significance of the taxon should not be ignored . A case of bronchitis in a dog which was simultaneously colonized by three different strains of Pasteurella is described . Also septicaemic pasteurellosis in a Nile crocodile (Crocodylus niloticus) is reported and for the first time prevalence of various serotypes in pasteurellosis of animals in Zimbabwe.

Kansenshogaku Zasshi, 1994 Feb, 68(2), 242 - 8
{Three cases of Pasteurella multocida infection in the respiratory tract}; Inoue Y et al.; Pasteurella multocida (P . multocida) is well recognized as "normal flora" in the upper respiratory tract of cats, dogs and other animals . Recently, various infections due to P . multocida in human have been noted as pulmonary infections in the patients with chronic pulmonary diseases as well as skin abscesses or septicemia after an animal bite or scratch . We report here three cases of respiratory tract infections caused by P . multocida . The first two patients had acute exacerbation of bronchiectasis caused by P . multocida and the other patients with pulmonary emphysema developed pneumonia . These three patients improved by antibiotic therapy . In Japan, P . multocida respiratory tract infection is rare, but it may become more common in the future . Therefore, it seems to be important to take this pathogen into consideration in the management of chronic lung disease.

Dtsch Tierarztl Wochenschr, 1994 Feb, 101(2), 75 - 7
{The effect of different flavophospholipol levels in rabbit feed on the fattening rate of hybrid rabbits}; Hartmann J et al.; The present study was designed to evaluate the effects of dietary flavophospholipol ("Bambermycin") levels on the growth performance in 400 hybrid rabbits from weaning at 28 days until slaughter at 74 days of age . The fryers were randomly assigned to one of the four dietary treatments that consisted of either 0, 4, 8 or 16 ppm supplemental flavophospholipol . Each treatment was applied to 100 animals each . Dietary treatments exerted no significant effects on live weight, daily weight gain and feed consumption across the entire fattening period . However, daily weight gains in treated fryers were increased by 2.4% on the average . The only significant treatment effect was found in feed conversion (food/gain) in fryers fed 16 ppm supplementation; kilogram food intake per kilogram weight gain was reduced by 0.16 kg as compared to the control group . Mean feed conversion in treated groups was improved by 3.5% on the average . In addition of flavophospholipol a significantly decreased mortality was observed . It was remarkable, that losses caused by infection in control were exclusively due to infections by Bordetella species, while the treated groups were only caused by Pasteurella species.

Infect Immun, 1994 Feb, 62(2), 579 - 88
Analysis of toxinogenic functions associated with the RTX repeat region and monoclonal antibody D12 epitope of Escherichia coli hemolysin; Rowe GE et al.; Amino acids (aa) 550 through 850 of the Escherichia coli hemolysin (HlyA) contain sequences important for several steps in cytolysis . These include the Ca(2+)-binding glycine-rich tandem repeats recognized by the monoclonal antibody A10, the putative HlyC-dependent acylation site that corresponds to the monoclonal antibody D12 epitope, and the erythrocyte specificity domain which confers erythrolytic activity to the Pasteurella haemolytica leukotoxin . To further investigate the toxinogenic functions associated with this region of HlyA, we constructed mutants in the hlyA sequences coding for the repeat region and the D12 epitope . Mutants were analyzed for anti-HlyA antibody reactivity, cytolytic activities, target cell binding, Ca2+ requirements, and virulence . The D12 epitope was mapped to aa 673 through 726, with portions of the epitope both amino terminal and carboxy terminal to aa 700 . This region was necessary, but not sufficient, for toxin binding to erythrocytes . A substitution at aa 684 resulted in loss of the D12 epitope, while cytolytic activity was retained . The nature of the D12 epitope and its associated functions are discussed . The A10 epitope mapped to residues 745 through 829, corresponding to repeats 4 through 11 . Insertions within the glycine-rich repeats resulted in mutant forms of HlyA which retained A10 reactivity but required increased Ca2+ for lytic activity . These in vitro effects on cytolysis corresponded to a significant decrease in HlyA-mediated virulence in mice . HlyA from one insertion mutant was able to associate with leukocyte membranes under conditions that were Ca2+ deficient for cytolysis . The role of the glycine-rich repeats and Ca2+ in HlyA activity are discussed.

FEMS Microbiol Lett, 1994 Jan 15, 115(2-3), 253 - 7
Binding of bovine transferrin by Pasteurella multocida serotype B:2,5, a strain which causes haemorrhagic septicaemia in buffalo and cattle; Veken JW et al.; Pasteurella multocida serotype B:2,5, which causes haemorrhagic septicaemia in buffalo and cattle, was examined for the presence of transferrin-binding proteins . An 82-kDa iron-regulated outer membrane protein was found which specifically binds bovine transferrin . In contrast, P . multocida serotype B:3,4, associated with haemorrhagic septicaemia in feral ruminants, did not express transferrin-binding proteins . These results might indicate a role for transferrin binding in the pathogenesis of haemorrhagic septicaemia in cattle and buffalo.

J Immunol Methods, 1994 Jan 3, 167(1-2), 35 - 45
Evaluation of different methods for the detection of outer membrane proteins and lipopolysaccharides of Pasteurella haemolytica by immunoblotting; Davies RL et al.; The optimal conditions for the detection of outer membrane proteins (OMPs) and lipopolysaccharide (LPS) of Pasteurella haemolytica by immunoblotting were evaluated . The variables examined included the equilibration time of the gels before transfer, composition of the transfer buffer, type of blotting membrane, blocking agent, effect of the zwitterionic detergent Empigen BB on protein renaturation, and the development reagent . The composition of the transfer buffer and time of gel equilibration significantly affected the efficiency of transfer of both OMPs and LPS . However, the optimal conditions for the transfer of OMPs were not the same as those for LPS . Thus, optimal transfer of OMPs occurred in Tris-glycine buffer, with prior equilibration of the gels to allow for expansion, whereas optimal transfer of LPS was achieved in Tris-glycine-methanol buffer with no equilibration of the gels . In Tris-glycine-methanol buffer, gel equilibration resulted in a significantly reduced transfer of both OMPs and LPS, probably due to the removal of SDS from these components . The use of Zeta-Probe blotting membrane which, unlike nitrocellulose, does not require methanol for optimal protein binding, did not result in improved binding of OMPs or LPS in the absence of methanol and, even after prolonged blocking (> 2 h), gave higher background staining than did nitrocellulose . Effective blocking of nitrocellulose was achieved with 3% (w/v) gelatin, 2.5% (w/v) skimmed milk or 0.3% (v/v) Tween 20, whereas increased background staining occurred with 1% (w/v) bovine serum albumin or 1% (w/v) ovalbumin . The incorporation of Empigen BB in the primary antibody buffer did not improve antibody recognition of proteins as a result of their renaturation . For the horseradish-peroxidase enzyme development system, the substrate 3,3'-diaminobenzidine tetrahydrochloride was more sensitive, and developed more quickly, than 4-chloro-1-naphthol, but faded more rapidly after drying of the membrane . 4-chloro-1-naphthol was more suitable for identifying OMPs because less background staining occurred, whereas 3,3'-diaminobenzidine tetrahydrochloride was more suitable for the detection of LPS due to its greater sensitivity.

J Basic Microbiol, 1994, 34(1), 61 - 3
Iron supply of Pasteurella multocida and Pasteurella haemolytica; Reissbrodt R et al.; Pasteurella multocida and Pasteurella haemolytica do not produce hydroxamate- or phenolate type siderophores . However, transport- and utilization systems could be detected for the well known siderophores ferrioxamine B, E, G, rhizoferrin and the intermediate 2,3-dihydroxybenzoic acid by means of cross-feeding tests in both Pasteurella species . Enterobactin and ferrichrome did not feed any of the Pasteurella strains tested . Additionally, alpha-ketoacids and alpha-hydroxyacids such as pyruvic acid, alpha-hydroxyisovaleric acid and others acting as primary metabolites enabled growth of P . multocida and P . haemolytica under iron limitation.

Burns, 1994, 20 Suppl 1, S27 - 30; discussion S30-1
Use of glycerol-preserved skin in plastic surgery; Wai RT; It is not always possible to effect immediate closure of wounds and patients' wounds are therefore exposed to risks of desiccation and infection . In the Plastic Surgery Department, Zuider Ziekenhuis, Rotterdam, we have used glycerolized allogeneic donor as a biological dressing in four patients; a polytrauma patient with a compound fracture of the left lower leg; a diabetic with necrotizing fasciitis; a patient with a cat bite on the lower leg, infected with Pasteurella multocida, and a child with large congenital naevi . On clinical grounds there are suggestions of a correlation between the degree of contamination and vascularization of the wound bed and graft take . Furthermore, our experience with the sandwich technique after excision of a large congenital naevus was positive.

Poult Sci, 1994 Jan, 73(1), 7 - 17
Differences in major histocompatibility complex frequencies after multitrait, divergent selection for immunocompetence; Kean RP et al.; White Leghorn chickens from lines selected for four immune-response traits (IR lines) were serotyped for B system alloantigens characterizing the haplotypes and genotypes to examine the effect of divergent selection for multitrait immunocompetence on MHC haplotype and genotype frequencies . The selected lines were derived from the Ottawa Strain 7 . The selection index included four immunocompetence traits: antibody production against Mycoplasma gallisepticum (MG) and Pasteurella multocida, inflammatory response to phytohemagglutinin, and reticuloendothelial carbon clearance . The four lines include two replicates of high and low multitrait-immunocompetence lines . After four cycles of selection, significant differences (P < .05) in several B system haplotype frequencies were observed, both among IR lines and between the IR lines and the Ottawa Strain 7 . The B2 haplotype frequency was greater in all IR lines than in the Ottawa Strain 7 . The B21 frequency was less in both high lines than in the Ottawa Strain 7 . In comparisons among lines, frequencies of B21 were greater in both replicates of the low lines and the B12 and B19 frequencies were significantly greater (P < .05) in the high lines . A gene substitution model showed effects (P < .10) of specific haplotypes on MG and on the index . The B2 haplotype had a positive effect associated with MG . Haplotype B21 was positively associated with the multitrait index . Haplotype B13 had a negative effect on both MG and the index . Significant differences (P < .01) in genotype frequencies were also noted among the IR lines . Associations between specific MHC haplotypes or genotypes and immune-response traits may offer insight into MHC-mediated mechanisms of disease resistance.

Poult Sci, 1994 Jan, 73(1), 18 - 32
Direct and correlated responses to multitrait, divergent selection for immunocompetence; Kean RP et al.; Leghorn lines had been selected for an immunocompetence index based on four traits: antibody production to Mycoplasma gallisepticum (MG) and Pasteurella multocida (PM) vaccines, reticuloendothelial clearance of colloidal carbon (CCA), and cell-mediated, wing web response to phytohemagglutinin (PHA) . The purpose of this study was to produce replicated lines of chickens with divergent levels of multitrait immunocompetence by index selection . The objectives of analyses of Generations 5 to 7 of this study was to characterize these lines with respect to immune-response traits, correlations among these traits, and correlated responses in other important production traits . Differences (P < .05) existed between the lines selected for high or low immune response and between the two replicates in mean breeding values and in individual immune-response traits . Averages of heritability estimates, weighted by number of offspring and pooled across three generations (two cycles of selection), estimated by using sire variance components and parent-offspring correlations were, respectively, .16 and .09 for the index, .31 and .08 for MG, .21 and -.02 for PM, .06 and .05 for CCA, and .08 and .12 for PHA . Realized heritabilities (response divided by effective selection differential) pooled across the two selection cycles, were .19 and .11 for the index, .06 and -.01 for MG, .44 and .32 for PM, 1.52 and -1.21 for CCA, and .48 and .15 for PHA, for Replicates 1 and 2, respectively . Phenotypic correlations among traits were generally small, and several estimates were negative . Estimates of genetic correlation varied widely . Juvenile and adult body weights, age of first egg, 32-wk egg weight, and rate of egg production were analyzed to evaluate effects of selection on these traits of direct economic importance . Very few differences were noted.

Lab Anim, 1994 Jan, 28(1), 19 - 25
Comparison of indirect haemagglutination test, gel-diffusion precipitin test, and enzyme-linked immunosorbent assay for detection of serum antibodies to Pasteurella multocida in naturally and experimentally infected rabbits; Kawamoto E et al.; Enzyme-linked immunosorbent assay (ELISA), gel-diffusion precipitin test (GDPT), and indirect haemagglutination test (IHAT) were evaluated for the detection of antibodies to Pasteurella multocida in both naturally and experimentally infected rabbits . A total of 285 rabbit serum samples from 7 rabbit colonies were tested by ELISA, GDPT, and IHAT, and nasal cultures were taken coincidentally to use as the standard in the serological tests . There was better correlation (98.0%) between the results of ELISA and positive nasal culture than between the GDPT (86.3%) or IHAT (23.5%) and positive nasal culture . In addition, ELISA and GDPT were positive in 26 (11.1%) and 21 (9.0%) of 234 serum samples from nasal culture negative rabbits, respectively . In experimentally infected rabbits, antibodies detected by the ELISA and GDPT began to rise one to 3 weeks post-inoculation . IHAT did not detect antibodies . These results are discussed in terms of value to serodiagnosis of rabbit pasteurellosis.

J Wildl Dis, 1994 Jan, 30(1), 16 - 9
Effects of modified Cary and Blair medium on recovery of nonhemolytic Pasteurella haemolytica from Rocky Mountain bighorn sheep (Ovis canadensis canadensis) pharyngeal swabs; Wild MA et al.; Modified Cary and Blair transport medium (MCB) was evaluated for recovery of Pasteurella spp . from pharyngeal swabs of healthy Rocky Mountain bighorn sheep (Ovis canadensis canadensis) . In experiment one, three pharyngeal swabs were collected from each of 25 bighorns . Pasteurella haemolytica was recovered from 21 of 25 swabs tested almost immediately and from 16 of 25 swabs held in MCB medium at about 22 C for 24 hr before testing (P > 0.10) . Recovery of P . haemolytica decreased (P < 0.005) to 1 of 25 when swabs were held in MCB medium at about 22 C for 48 hr before testing . In experiment two, four pharyngeal swabs were collected from each of ten bighorns and held in MCB medium at about 5 C for < or = 5, 24, 48, or 72 hr prior to testing . Recovery was unaffected by storage at 5 C; P . haemolytica was isolated from all 40 of these samples . All Pasteurella spp . isolates were nonhemolytic P . haemolytica . In experiment one, most isolates were serotype 4; in experiment two, serotype 3 was most common . We propose that MCB medium is effective for transporting bighorn sheep pharyngeal swabs for P . haemolytica screening because it imposes minimal or no effect on recovery when held < or = 24 hr at 22 C or < or = 72 hr at 5 C.

J Wildl Dis, 1994 Jan, 30(1), 1 - 7
Bacteria isolated from nasal and tonsillar samples of clinically healthy Rocky Mountain bighorn and domestic sheep; Queen C et al.; Nasal and tonsillar samples were collected from 14 free-ranging clinically healthy Rocky Mountain bighorn sheep (Ovis canadensis canadensis) and 10 domestic sheep (Ovis aries) . We identified 194 bacterial isolates, including 101 from bighorn and 93 from domestic sheep . Of these isolates, 115 were gram-positive and 79 were gram-negative . Staphylococcus species were the most numerous gram-positive organisms and had a higher incidence in samples from domestic than from bighorn sheep . In contrast Streptococcus species were present in higher numbers in samples from bighorn sheep . Pasteurella haemolytica, the most common gram-negative bacterium, was isolated from five of five tonsillar but from none of ten nasal samples of domestic sheep, and from seven of eight tonsillar and three of ten nasal samples of bighorn sheep . Most bacteria isolated were considered opportunistic pathogens . However, of the bacteria isolated, P . haemolytica, P . multocida, and Actinomyces pyogenes are most frequently associated with respiratory disease.

Can J Vet Res, 1994 Jan, 58(1), 31 - 5
Passive immunity to Pasteurella haemolytica A1 in dairy calves: effects of preparturient vaccination of the dams; Hodgins DC et al.; Dairy cows from five herds were assigned to receive a commercial Pasteurella haemolytica vaccine or no vaccine at all, administered at six and three weeks before parturition . Vaccination was associated with increased leukotoxin neutralizing serum antibody titers in the dams (p < 0.001), and with increased titers in colostrum (p < 0.001) . Vaccination of dams also had a significant association with increased passive leukotoxin neutralizing antibody titers in their calves (p < 0.001) . Vaccination was also associated with increased indirect agglutinating antibody titers in serum of the dams (p < 0.001) . In the analysis of agglutinating antibody titers in colostral whey the interaction "vaccination*herd" was found to be significant (p < 0.001), indicating that the effects of vaccination on colostral titers were not consistent from herd to herd . The analysis was repeated, stratifying by herd . Vaccination was associated with increased agglutinating antibody titers in colostrum (p < 0.05) in three herds of the five in the study . In two of these three herds there were significant increases in passive neonatal titers associated with vaccination . In the remaining herd the mean IgG1 level in the calves was consistent with failure of passive transfer of immunoglobulins (IgG1 < 8.0 g/L) . These results suggest that preparturient vaccination of dairy cows can induce modest increases in passive antibody titers to antigens of Pasteurella haemolytica in their calves, but the antigen of interest and the population being studied can affect the outcome.

Can J Vet Res, 1994 Jan, 58(1), 25 - 30
Experimental model of swine pneumonic pasteurellosis using crude Actinobacillus pleuropneumoniae cytotoxin and Pasteurella multocida given endobronchially; Chung WB et al.; This study was designed to develop and characterize a swine pneumonic pasteurellosis model by concurrent introduction of Pasteurella multocida type A and Actinobacillus pleuropneumoniae crude cytotoxin . After a series of preliminary experiments, a combination of 4 x 10(9) P . multocida and 4,000 toxic units of A . pleuropneumoniae crude cytotoxin was determined to produce optimal results . A total of 48 pigs were divided into four groups of 12 pigs each . The control group received buffered saline only . Four pigs from each group were randomly selected for necropsy 3, 7 and 14 days postinoculation (PI) . Inoculation of pigs with P . multocida and A . pleuropneumoniae cytotoxin (group 1) resulted in moderate to severe pneumonia . Pasteurella multocida was isolated from pneumonic lesions, grossly normal lung, and bronchial lymph nodes of all group 1 pigs throughout the 14 day experimental period . Pathological changes typical of field cases of swine pneumonic pasteurellosis were produced . Pigs inoculated with P . multocida alone (group 2) had pneumonic lesions and P . multocida was reisolated from lungs at three days PI . Pasteurella multocida was not isolated from these pigs at 7 and 14 days PI, except for one pig in which an abscess developed in the thorax . Pulmonary lesions induced by A . pleuropneumoniae crude cytotoxin alone (group 3) were transient and resolved by seven days PI . Group 1 pigs had significantly greater lung lesion volumes than group 2 and 3 pigs at 3, 7 and 14 days PI . Statistical analysis indicated a significant interactive effect of P . multocida and A . pleuropneumoniae cytotoxin on the development of lung lesion volumes at 7 and 14 days PI (p < 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)

Vet Res, 1994, 25(1), 8 - 15
Pasteurella multocida: oropharyngeal carriage and antibody response in breeders; Donnio PY et al.; The presence of Pasteurella multocida in the oropharynx of 58 pig, cattle or rabbit breeders whose livestock had suffered from pasteurellosis was investigated using a selective medium . Blood samples for serological studies were collected at the same moment . Nineteen breeders were found to host one P multocida subsp multocida strain . Oropharyngeal carriage of P multocida was found to be more frequent in pig breeders (42% of individuals) than in cattle (10%) or rabbit (0%) breeders . Genomic polymorphism among 10 D2 strains was found by restriction endonuclease analysis using pulsed-field gel electrophoresis (REA-PFGE) . Antibodies to P multocida were found in the sera of 32 of these 58 breeders, whereas only 2 of the 70 controls had antibodies . These results, recorded from healthy individuals, show that P multocida, acting as an opportunistic bacterium, may be responsible for occupational diseases . Nevertheless, the strong prevalence of specific antibodies makes the presence of antibodies in the sera of these breeders an insufficient indication of a current patent infection.

Am J Vet Res, 1994 Jan, 55(1), 49 - 54
Intranasal administration of Pasteurella multocida toxin in a challenge-exposure model used to induce subclinical signs of atrophic rhinitis in pigs; van Diemen PM et al.; A challenge-exposure model was developed for dose-dependent induction of subclinical (moderate) atrophic rhinitis (AR) in conventionally raised Dutch Landrace and Large White pigs, about 4 weeks old . Under favorable climatic and housing conditions, pigs were intranasally challenge-exposed with Pasteurella multocida-derived toxin (Pm-T) 3 days after pretreatment by inoculation with 1% acetic acid . Pigs were challenge-exposed with 1 of the following Pm-T doses: 0 (control), 5, 13, 20, or 40 micrograms of Pm-T/ml of phosphate-buffered saline solution (PBSS), 0.5 ml/nostril/d on 3 consecutive days . Five weeks after challenge exposure, subclinical (moderate) AR status was defined as intermediate conchal atrophy (grade 2 for ventral conchae on a 0 to 4 scale and grade 1 or 2 for dorsal conchae on a 0 to 3 scale, respectively) and perceptible difference in change in brachygnathia superior (cBS) between control and challenge-exposed pigs between the beginning and end of the study . All Pm-T-exposed pigs had nasal damage that was dose-dependent . The higher Pm-T doses resulted in higher ventral conchae atrophy and dorsal conchae atrophy scores . The cBS increased with applied Pm-T dose, resulting in significant (P < 0.05) differences between controls (3.88 mm) and the 13-, 20-, and 40-micrograms Pm-T-treated groups (7.77, 6.58, and 7.98 mm, respectively) . In response to the applied dose, weight gain per week for Pm-T-exposed pigs was lower than that of controls after week 3 (P < 0.01).(ABSTRACT TRUNCATED AT 250 WORDS)

Vet Pathol, 1994 Jan, 31(1), 55 - 60
Tissue factor expression in bovine endothelial cells induced by Pasteurella haemolytica lipopolysaccharide and interleukin-1; Breider MA et al.; Pasteurella haemolytica in cattle produces fibrino-hemorrhagic pleuropneumonia characterized by extensive pulmonary microvascular thrombosis and parenchymal necrosis . The purpose of this in vitro study was to determine if P . haemolytica lipopolysaccharide (LPS) promotes vascular thrombosis by inducing a procoagulant state in vascular endothelial cells . After treatment of confluent monolayers of bovine pulmonary artery endothelial cells with various concentrations of either P . haemolytica LPS or Escherichia coli LPS, the procoagulant activity of the endothelial cells was determined using a chromogenic assay dependent on cellular tissue factor expression . The LPS treatment induced significant increases in cellular tissue factor expression in a LPS concentration- and time-dependent manner . Highest levels of tissue factor were present at 22 hours after treatment, although high LPS concentrations induced moderate tissue factor levels at 5 hours after treatment . Interleukin-1 also induced tissue factor expression in endothelial cells and enhanced the LPS-induced effects . This interleukin-1 effect could be diminished by concurrent use of an interleukin-1 receptor antagonist . These results demonstrate that LPS and cytokine promotion of a procoagulant state in endothelial cells occurs in vitro . Similar mechanisms may play a role in P . haemolytica-mediated pulmonary vascular thrombosis.

Dtsch Tierarztl Wochenschr, 1994 Jan, 101(1), 27 - 30
{Detection of toxin-producing strains of Pasteurella multocida in nasal and tonsil swabs, a possibility for the control of atrophic rhinitis in swine breeding herds? Parallel comparison with other research methods}; Matschullat G et al.; Nasal, and partly tonsil swabs from pigs have been investigated by cell culture and ELISA respectively on toxigenic Pasteurella multocida strains . The controlled animals are from 89 non atrophic rhinitis- and 42 a.r.-positive herds . Toxin forming strains could not be isolated in non suspicious herds . 23 out of 42 infected herds showed toxigenic Pasteurellae . Despite a high increase in sample numbers, Pasteurella could not be detected in 19 infected herds (45%) . When diagnosing atrophic rhinitis, we thus recommend to rely only on positive results of isolation . In all other cases, all other methods will have to be used in combination . Further research is necessary to find safe and fast test methods . According to recent experience, the blood-serological toxin screening test can be considered useful.

Berl Munch Tierarztl Wochenschr, 1994 Jan, 107(1), 15 - 9
{Detection of atypical toxin-producing Pasteurella strains on the nasal mucosa of calves and their differential diagnostic limits for Pasteurella haemolytica and Pasteurella multocida}; Kruger M et al.; The bacterial colonisation on nasal mucous membranes of calves at age of 3, 5 to 5 months was investigated by cotton swabs . Three Pasteurella species were found (P . multocida subspecies multocida, P . haemolytica, P . avium) as monocausal infection as well as pluricausal infection . P . avium was characterised by colonial morphology, bacterioscopy, biochemical properties, polypeptide-pattern in SDS-PAGE and by electronmicroscopic investigation and was differentiated from P . multocida and P . haemolytica . It is very interesting, that the P . avium strains possess an antigen structure, reacting with monoclonal antibodies directed against the heat labile-toxin of P . multocida subspecies multocida.

Vet Rec, 1994 Jan 1, 134(1), 5 - 7
Antimicrobial susceptibility of Actinobacillus pleuropneumoniae, Pasteurella multocida and Salmonella choleraesuis isolates from pigs; Raemdonck DL et al.; The in vitro susceptibility of 839 isolates of Actinobacillus pleuropneumoniae, 969 isolates of Pasteurella multocida and 104 isolates of Salmonella choleraesuis from pigs to the fluoroquinolone danofloxacin, and eight other commonly used antimicrobial drugs was determined by veterinary diagnostic laboratories in Europe, Japan, South Africa and North America between 1989 and 1991, by using a broth microdilution technique . The minimum inhibitory concentrations of danofloxacin, amoxycillin, ceftiofur, erythromycin, gentamicin, lincomycin, oxytetracycline, spectinomycin and trimethoprim:sulphamethoxazole (ratio 1:19) that prevented the growth of 90 per cent of the bacteria were 0.125, < or = 0.5, < or = 0.125, 8, 8, 32, 32, 64 and < or = 0.25 microgram/ml for A pleuropneumoniae, 0.06, 1, < or = 0.125, 8, 4, 64, 8, 32 and 8 micrograms/ml for P multocida, and 0.125, > 64, < or = 1, > 64, 1, > 64, > 64, 64 and < or = 0.25 microgram/ml for S choleraesuis . These data confirm the high in vitro potency of danofloxacin against field isolates that show significant resistance to several other antibacterial drugs.

J Clin Microbiol, 1994 Jan, 32(1), 256 - 8
Characterization of Escherichia coli isolated from the tonsils of cattle; Frank GH et al.; During our studies on tonsillar colonization by Pasteurella haemolytica, we consistently found Escherichia coli to be one of the most prominent and prevalent bacterial species in the tonsils of healthy cattle . Since tonsillar isolates have not been characterized, we grouped 124 isolates from 87 healthy cattle from eight sources by hemolytic zone size and by carbon source utilization and probed them for selected virulence genes . They formed 3 groups by hemolytic zone size and 18 groups (of 2 to 31 isolates) by their metabolic patterns . Most groups included isolates from more than one source . Two isolates contained the Shiga-like toxin gene, and nine others contained the F41 accessory gene.

J Am Vet Med Assoc, 1994 Jan 1, 204(1), 102 - 7
Interaction of Mycoplasma hyopneumoniae and Pasteurella multocida infections in swine; Amass SF et al.; To investigate the interaction between Mycoplasma hyopneumoniae and Pasteurella multocida infection, 32 pigs were randomly assigned by litter, sex, and weight to 4 treatment groups . Group-1 pigs were inoculated with M hyopneumoniae and allowed to recover from M hyopneumoniae infection . Group-2 pigs were vaccinated against M hyopneumoniae and then inoculated with M hyopneumoniae . Group-3 pigs were inoculated with M hyopneumoniae and developed clinical signs of mycoplasmosis . Group-4 pigs had never been exposed to M hyopneumoniae . All pigs were initially seronegative for M hyopneumoniae . All pigs were subsequently inoculated with P multocida and euthanatized 2 weeks later . Pasteurella multocida was isolated only from the lungs of group-3 pigs, and these pigs had a significantly higher median percentage of lung surface area affected by pneumonia than did pigs in the other groups . For group-3 pigs, percentage of lung surface area affected by pneumonia was positively correlated with the number of P multocida colonies isolated . We concluded that P multocida is not a primary respiratory pathogen in pigs, but that M hyopneumoniae infection can render the lungs susceptible to P multocida colonization and infection . Pigs recovered from or vaccinated against infection with M hyopneumoniae were resistant to P multocida infection.

Appl Environ Microbiol, 1994 Jan, 60(1), 180 - 6
Evidence of a dormant but infective state of the fish pathogen Pasteurella piscicida in seawater and sediment; Magarinos B et al.; The stability of Pasteurella piscicida strains in seawater and sediment microcosms at different temperatures (6 and 20 degrees C) was investigated during a 1-month period . Three strains of P . piscicida showed similar survival kinetics . By a standard plate count method they survived in water and sediment for only 6 to 12 days, depending on the strain and type of microcosm . During this starvation period, the metabolic activity of the cells was reduced by more than 80% . Culturable cells of each P . piscicida strain persisted better in sediment than in water, as well as at 20 degrees C compared to 6 degrees C . However, in all the microcosms, the acridine orange direct counts remained at about 10(5) cells per ml during the experimental period, which demonstrated that P . piscicida possesses a capacity to enter a viable but not culturable state . Moreover, dormant cells were always resuscitated by the addition of fresh medium to the microcosms, since we recovered numbers of culturable cells similar to the acridine orange direct counts . These resuscitated cells exhibited the same respiration rate as that seen prior to the start of the experiments . Although the biochemical, physiological, and serological characteristics; lipopolysaccharides; membrane proteins; and plasmid content of P . piscicida strains were unaffected during the starvation conditions, the dormant cells were smaller (dwarf cells) and had increased surface hydrophobicity . The starved cells maintained their infectivity and pathogenic potential for fish, with 50% lethal doses similar to those of the original strains.

Microbiol Immunol, 1994, 38(1), 31 - 8
The transposon-like structure of IS26-tetracycline, and kanamycin resistance determinant derived from transferable R plasmid of fish pathogen, Pasteurella piscicida; Kim EH et al.; Tetracycline (pp-tet), and kanamycin (pp-kan) resistance genes were cloned from a transferable R plasmid of fish pathogen Pasteurella piscicida, and complete nucleotide sequences were determined . The pp-tet was a class D Tet determinant constructed with the tetA resistance gene of 1,182 bp encoding a protein with a deduced molecular mass of 41 kDa and the tetR repressor gene of 654 bp encoding a product of 24 kDa . The pp-tet was highly homologous to the tet(D) of plasmid RA1 isolated from Aeromonas hydrophila with two nucleotide differences in the tetR, and of plasmid pIP173 from Salmonella ordonez with two nucleotide differences in the tetA . The pp-kan contained 813 bp encoding a 31 kDa protein of 271 amino acids, and was classified into type aph-Ic . It was identical to the aphA7 in the IAB operon of pBWH77, in which was originally found an isolate of Klebsiella pneumoniae, in its nucleotide sequences and hybrid promoter construction . The genes were connected by an insertion sequence IS26 of 820 bp, and were flanked by repeated copies in direct orientation at the 3' flanking region of the pp-tetA and in inverted orientation at the 3' flanking region of the pp-kan . The genetic elements are organized like a complex transposon by close linkage of the IS26 and the pp-tet and -kan.

Biochimie, 1994, 76(1), 9 - 14
Cloning and expression of two Pasteurella multocida genes in Escherichia coli; Manoha F et al.; A library of cloned Pasteurella multocida (toxigenic strain 9222, serotype D2) genomic sequences was constructed in Escherichia coli by incorporating TaqI digestion fragments into the plasmid vector pUC19 . Immunological screening with antibodies directed against porin H, the major protein of the P multocida outer membrane, allowed the identification of a recombinant plasmid containing a 2.9-kbp DNA insert . This plasmid encoded the synthesis of two polypeptides, p25 (25 kDa) and p28 (28 kDa) which were detected in the different compartments of the E coli transformant . The peptide p25 was more abundant in the periplasm whereas p28 was mainly found in the cell envelope and in the cytosol . Immunological analysis indicates that p25, in contrast to p28, is antigenically related to porin H of P multocida . The expression in E coli of the gene encoding p28 was enhanced by induction of the lac promoter.

Vet Microbiol, 1994 Jan, 38(3), 255 - 62
Administration of a bolus formulation of baquiloprim and sulphadimidine to calves: plasma concentration--time profiles and efficacy in suppressing experimental pneumonic pasteurellosis; Dassanayake L et al.; After administration of a bolus containing 0.8 g baquiloprim and 7.2 g sulphadimidine to six calves, plasma concentrations of these compounds exceeded MICs for Pasteurella haemolytica from about 3 hours to at least 48 hours . Experimental infection of three calves with 2.8 x 10(10) P . haemolytica type A1 organisms by the endobronchial route resulted in extensive pneumonic lesions, and smaller, similar lesions were found post mortem in two other calves which had received the same inoculum . Nine similar calves were also infected in the same manner after dosing six hours previously with a bolus providing doses of 4 mg/kg baquiloprim and 36 mg/kg sulphadimidine . Four of these calves received a second bolus 48 hours later . In contrast to the undosed calves, the treated calves exhibited few symptoms of respiratory disease following infection, and their temperatures remained normal . The mean temperatures of the control calves were significantly elevated for several days after infection . At post mortem examinations 6 days after infection, lung lesions were insignificant in all the dosed calves . P . haemolytica was isolated from the lungs of all 5 control calves but not from any treated animal . The combined dose of 40 mg/kg baquiloprim and sulphadimidine was the minimum recommended for use of the bolus in the field, and the results therefore predict efficacy when used in outbreaks of respiratory infection in calves associated with this organism.

Avian Dis, 1994 Jan-Mar, 38(1), 72 - 7
Invasion of epithelial cell monolayers by turkey strains of Pasteurella multocida; Lee MD et al.; Two serotype 3,4:A strains of Pasteurella multocida that differ in virulence in turkeys were examined for their ability to invade epithelial cell monolayers grown in tissue culture . Both organisms were comparably adherent to cells of turkey kidney origin . However, the virulent strain (86-1913) penetrated primary turkey kidney epithelial cell monolayers at 10 times the level of the low-virulence vaccine strain . The virulent strain was also able to invade porcine epithelial cells (PK15) and feline epithelial cells (CRFK) in cell culture . Neither organism invaded rabbit epithelial cells (RK13) . Invasion of turkey cells was prevented by inhibition of bacterial protein or RNA synthesis but not by pretreatment of the monolayers with periodate, trypsin, or neuraminidase . Invasion might be a mechanism of pathogenicity for this organism, contributing to colonization or virulence.

Avian Dis, 1994 Jan-Mar, 38(1), 33 - 6
Effect of genetic selection for increased body weight and sex of poult on antibody response of turkeys to Newcastle disease virus and Pasteurella multocida vaccines; Sacco RE et al.; Primary and secondary antibody responses of 671 turkeys of two genetic lines to Newcastle disease virus (NDV) and Pasteurella multocida vaccines were examined . The randombred control line (RBC2) and a subline (F) of RBC2 had been selected for increased 16-week body weight . Poults were vaccinated at 6 and 12 weeks of age, and serum samples were collected 3 weeks after each vaccination . Antibody titers were determined using an enzyme-linked immunosorbent assay . Line F turkeys had significantly higher 9-week and 15-week serum antibody titers to NDV than line RBC2 . However, line RBC2 had significantly higher serum antibody titers to P . multocida at 15 weeks of age than line F . The 9-week and 15-week serum antibody titers to NDV were significantly higher in females than males, but males had significantly higher 15-week serum antibody titers to P . multocida than females . Sex of poults did not contribute significantly to variation in serum antibody response to P . multocida at 9 weeks of age.

Rev Elev Med Vet Pays Trop, 1994, 47(1), 19 - 20
{First case of an outbreak of hemorrhagic septicemia caused by Pasteurella multocida serotype B6 in northern Cameroon}; Martrenchar A et al.; For the first time in Cameroon, a strain of Pasteurella multocida serotype B6 was isolated from a haemorrhagic septicaemia outbreak in Zebu cattle in the area of Maga (Far Northern Province) . Through the mouse protection test, evidence was given that there was no cross protection between this strain and the strain of Pasteurella multocida serotype E6 which is used as an inactivated vaccine in Cameroon . Hence, it is recommended to use a combined vaccine including both serotypes for the Central and West African countries.

Vet Res Commun, 1994, 18(2), 133 - 40
Altered endothelium-dependent alpha-adrenergic vascular reactivity following exposure to Pasteurella haemolytica antigens; Weekley LB et al.; Rats were injected with sterile saline (controls), 10(5) cfu of Pasteurella haemolytica (biotype A) obtained from a commercial vaccine, or a commercial Pasteurella leukotoxoid vaccine . Three days after vaccination, the animals were killed and the thoracic aorta was removed . In some experiments the vascular endothelium was mechanically removed . Each isolated aorta was placed in a tissue bath and the biophysical responses to methoxamine (alpha-1 agonist) were determined . In separate experiments the endothelial surface was examined by scanning electron microscopy . In endothelium-intact vessels both vaccines appeared to enhance the contractile response to methoxamine . On the other hand, in endothelial-denuded vessels, the methoxamine-mediated contractile response was enhanced in the P . haemolytica-treated group but not in animals vaccinated with leukotoxoid . Furthermore, scanning electron microscopy revealed deposition of a fibrin-like material on the endothelial surface of vaccinated animals . These results suggest that exposure to vaccine-derived P . haemolytica antigens alters the morphology and adrenergic responsiveness of vascular smooth muscle.

Rev Latinoam Microbiol, 1994 Jan-Mar, 36(1), 57 - 66
{Evaluation of phagocytosis, bactericidal effect, and cytotoxicity of Pasteurella haemolytica and Pasteurella multocida in bovine alveolar macrophages}; Morales JF et al.; Evaluation of phagocytosis and bactericidal effect of Pasteurella haemolytica and P . multocida was conducted on bovine alveolar macrophages freshly obtained through bronchioalveolar washings from live animals . Cytotoxic activity of these bacteria on the alveolar macrophages was evaluated through the simple visual assay in microplates, using bovine blood leukocytes as a comparative target cell . In order to evaluate phagocytosis the following variables were considered P . haemolytica and P . multocida (independently) in contact with alveolar macrophages, P . haemolytica and P . multocida in suspension as a positive control of bacterial growth, and RPMI-1640 medium alone, as a negative control of bacterial growth . To measure bactericidal capacity, bacteria were incubated with plastic adhered alveolar macrophages at 30 minutes and 3 hours intervals . Samples incubated 30 minutes were taken as phagocytosis-base readings and at the 3 h interval to evaluate bactericidal capacity of the alveolar macrophages on phagocytized bacteria . Reading of the samples of each evaluation was conducted in a spectrophotometer a 380 nm . Phagocytosis results indicated that bacterial proliferation was higher when bacteria were alone as compared when they were with alveolar macrophages (p < 0.05) . Bactericidal capacity of the macrophages was efficient because bacterial numbers were higher in the first evaluation as compared to the second (p < 0.05) . It was demonstrated that the cytotoxic effect of P . haemolytica was more severe on blood leukocytes as compared to alveolar macrophages (p < 0.05) . There was no evidence of P . multocida cytotoxicity on the evaluated cells . With the development of this technique for the obtention of alveolar macrophages and using spectrophotometry for the phagocytosis and bactericidal effect evaluations, numerous variables and samples can be tested, such as opsonized bacteria or to measure the behaviour of alveolar macrophages infected with different agents involved in the bovine pneumonic complex.

Arch Med Res, 1994 Summer, 25(2), 235 - 9
Mycoplasma hyopneumoniae: interaction with other agents in pigs, and evaluation of immunogens; Ciprian A et al.; Mycoplasmal pneumonia of swine causes considerable economic losses in porciculture . Diverse agents, such as environmental stress and infectious microorganisms, increase the possibility of infection, and the damage to pulmonary tissue when the infection is established . It is known that Mycoplasma hyopneumoniae has an important role in this disease, in addition to secondary microbial agents, such as Pasteurella multocida and Actinobacillus pleuropneumoniae . However, the characteristics of this disease in farms are well known . In this work we review the interactions among the different microorganisms involved and the immunological strategies utilized to control this disease . The interaction between Mycoplasma hyopneumoniae and P . multocida in experimental pneumonia was reported by us in conventional pigs . M . hyopneumoniae causes mild pneumonia, whereas P . multocida aggravates the pneumonia initiated by M . hyopneumoniae . The disease has been reproduced to test the efficacy of two immunogens, and can also be used to evaluate some antibiotics . A M . hyopneumoniae bacterine administered intraperitoneally conferred more protection than when it was used with adjuvant, although protection was not complete and colonization by M . hyopneumoniae was not prevented, as is claimed to have been the case with other vaccines.

Rev Elev Med Vet Pays Trop, 1994, 47(3), 289 - 90
Pasteurella haemolytica A2 infection in two sheep flocks in Tripoli area (Libya); Moustafa AM; 330 sheep from two different flocks were examined for respiratory manifestations including serous to mucopurulent nasal discharge dyspnoea, cough and light depression . 55 animals were sick (16.6%) and 13 died (3.9%) . Samples were taken from healthy, diseases and dead cases as well . Pasteurella haemolytica A2 was identified from 18 isolates, and serotyped by rapid plate agglutination (RPA) . Drug susceptibility was tested and treatment applied in line with the results.

Ann Dermatol Venereol, 1994, 121(5), 399 - 401
{Multocida Pasteurella and leg ulcer}; Chraibi A et al.; INTRODUCTION . We observed two cases of Pasteurella and discuss the role of bacterial sampling in ulcerations of the lower limb . CASE REPORTS . In the first case, Pasteurella was discovered in an ulceration of the lower limb and was cured with no particularly specific care . In the second case, Pasteurella had been inoculated by scratching an ulceration and could not be cured without specific treatment . DISCUSSION . No specific pathological consequence in chronic carriers could be possible, a situation which has often been reported in the literature . Pasteurella is an unusual specific cause of ulcerations of the lower limb.

Can J Vet Res, 1994 Jan, 58(1), 1 - 5
Pasteurella haemolytica leukotoxin induces histamine release from bovine pulmonary mast cells; Adusu TE et al.; Pasteurella haemolytica A1 leukotoxic culture supernatant was evaluated for its ability to induce histamine release from bovine pulmonary mast cells isolated by enzymatic dispersion of lung tissue . Histamine was measured by a radioimmunoassay technique . Leukotoxic culture supernatant of P . haemolytica significantly released histamine in a time and concentration-related manner . This effect was lost when culture supernatant was heat-inactivated or preincubated with leukotoxin neutralizing rabbit serum . Preincubation of the mast cells with propranolol or p-bromophenacyl bromide reduced the histamine-releasing effect of leukotoxin, while verapamil enhanced release . Experimental infection of calves with P . haemolytica A1 reduced the total histamine content of pulmonary mast cells recovered at postmortem . Histamine release induced by P . haemolytica leukotoxin is likely an important factor in the pathogenesis of bovine pneumonic pasteurellosis.

Dtsch Tierarztl Wochenschr, 1993 Dec, 100(12), 488 - 91
{Toxin-producing pasteurellas in a cattle herd}; Ehlers J et al.; Besides other agents, indole-negative Pasteurellae, producing dermonecrotic Pasteurella toxin, were isolated from the noses of calves in a herd with enzootic bronchopneumonia . In some blood sera, antitoxin was detected . According to their biochemical activities, isolated strains were classified as P . multocida ssp . septica (ornithine-negative), P . avium (biovar 2), and P . canis (biovar 2) . However, in DNA-DNA hybridisation tests there was much coincidence with P . multocida . In experimental calves, pneumonic lesions were produced with one of the isolates as well as with the dermonecrotic toxin . Therefore, indole negative toxinogenic Pasteurellae are considered pneumonia causing agents . They should be taken into account in bacteriological diagnostic and for production of herd specific bacterins.

Kansenshogaku Zasshi, 1993 Dec, 67(12), 1228 - 32
{Five cases of respiratory infection due to Pasteurella multocida}; Watanabe Y; Five cases of respiratory infection in which Pasteurella multocida (P . multocida) was isolated from sputum were reported . All of the five patients had animal contact, and three of the five patients suffered from bronchiectasis . No underlying pulmonary disease was found in one case, P . multocida was also isolated from her cat . Patients were treated with ST, MINO + PIPC, ABPC + CFT, SBTPC, AMK + MINO with improvement . Thirty cases of respiratory tract infection due to P . multocida including our cases have been reported in Japan.

Endocrinology, 1993 Dec, 133(6), 3054 - 7
Induction of Fos-like immunoreactivity in the rat brain following Pasteurella multocida endotoxin administration; Elmquist JK et al.; The functional neuroanatomy of the immune system link to the CNS was investigated by assessing neuronal activity with Fos immunohistochemistry following systemic lipopolysaccharide (LPS) administration . Two hours after LPS robust Fos-like immunoreactivity (Fos-IR) was observed in several nuclear groups in the brain including the paraventricular and supraoptic nuclei of the hypothalamus, central nucleus of the amygdala, and nucleus of the solitary tract . A similar but diminished pattern of Fos-IR was present at 6 hours and was absent 24 hours after LPS administration . Investigation of the functional neuroanatomy of the acute phase reaction could prove to be critical in enhancing the ability of individuals to combat insults such as tissue damage and inflammation . The central nervous system (CNS), particularly the hypothalamus, is intimately involved in the coordination of the various aspects of the acute phase reaction (reviewed in 1) . Understanding the functional neuroanatomy by which the brain responds to immune system challenges would greatly augment the ability to control the deleterious and enhance the beneficial aspects of the acute phase reaction . In this study we have used lipopolysaccharide (LPS or endotoxin) administration as an experimental model to study immune system activation . LPS is a complex glycolipid and a component of the outer membrane of most Gram-negative bacteria (2) . Administration of LPS has been demonstrated to induce the secretion of several proteins including interleukin-1 (IL-1), tumor necrosis factor (TNF), and interleukin-6 (IL-6; reviewed in 3) . Further, it has been hypothesized that LPS induction of IL-1 and TNF is the key event in the pathogenesis of Gram-negative bacterial septic shock syndrome (2) . Many recent studies have utilized immunohistochemistry for Fos, the product of the immediate early gene c-fos, as a marker of neuronal activation . Fos is a nuclear-binding protein that is expressed at increased levels in activated neurons (4) . Although the exact function of Fos in the CNS is still unknown, it is thought that Fos is transcribed after cellular stimulation as a means to convert a stimulus into long-term genetic action (for reviews see 5,6) . This study investigated the activation of the CNS by peripherally administered LPS isolated from the bacterium Pasteurella multocida . As a marker of neuronal activation, immunohistochemistry for the Fos protein was performed and image analysis was utilized to quantify the Fos induction in the CNS.

Infect Immun, 1993 Dec, 61(12), 5001 - 7
Molecular analysis of the leukotoxin determinants from Pasteurella haemolytica serotypes 1 to 16; Burrows LL et al.; All sixteen serotypes of Pasteurella haemolytica were shown to produce a leukotoxin protein which is immunologically related to the well-characterized serotype 1 leukotoxin . All of the leukotoxins were weakly hemolytic and were able to bind to BL-3 target cells . The leukotoxin determinants were characterized by Southern blot hybridization by use of the previously cloned serotype 1 determinant as the probe, and a number of distinct classes were identified . The leukotoxin determinants from serotypes 2, 3, and 11 were cloned . Nucleotide sequence analysis of the lktC and lktA genes of the serotype 3 and 11 determinants revealed nucleotide substitutions throughout the coding sequences . A comparison of the lktC and lktA genes and deduced proteins of serotypes 1, 3, and 11 showed that they are highly homologous.

J Vet Pharmacol Ther, 1993 Dec, 16(4), 446 - 53
Disturbances in ex vivo vascular smooth muscle responses following exposure to Pasteurella haemolytica vaccines; Weekley LB et al.; Rats were vaccinated with saline (control) or one of the two commercially available Pasteurella haemolytica vaccines Presponse or Precon-PH . Animals were killed 3 days later and thoracic aorta removed for evaluation of the ex vivo biophysical responses to carbachol (CCh) . In some experiments, vascular endothelium was mechanically removed . Vaccination of rats impairs the endothelial-dependent relaxation to CCh . In vessels with endothelium removed, the contractile response to CCh is converted into a relaxation following vaccination . Treatment of endothelial-denuded vascular rings ex vivo with methylene blue, a guanylate cyclase inhibitor, reduced the vaccination effect . Treatment of vascular rings with the superoxide dismutase inhibitor diethyldithiocarbamate, impairs the relaxant response of de-endothelialized vessels to CCh in Presponse vaccinated rats while enhancing the relaxation response of vessels from Precon-PH vaccinated rats . De-endothelialized vessels from vaccinated rats, but not control rats, relaxed in the presence of N-monomethyl-L-arginine (L-NMMA), a competitive inhibitor of nitric oxide synthetase . Furthermore, in the presence of L-NMMA, the relaxant response to CCh is significantly enhanced by Precon-PH but not Presponse . The normal relaxant response to hydrogen peroxide is converted into a contraction following vaccination . Results suggest that exposure to commercially available P . haemolytica vaccines alters vascular smooth muscle reactivity to CCh and that several independent pathways may be altered.

Vet Q . 1993 Dec;15(4):184.
Comparison between the minimal inhibitory concentration of tilmicosin and oxytetracycline for bovine pneumonic Pasteurella haemolytica isolates; Hartman EG et al.; The minimal inhibitory concentration (MIC) of tilmicosin and oxytetracycline was determined for Pasteurella haemolytica isolated from Dutch cattle in 1991 . The agar dilution method was used . Of the Pasteurella haemolytica strains examined, 96% (24) had a MIC for tilmicosin of 2 micrograms/ml or lower; in one strain the MIC was 8 micrograms/ml . The MIC for oxytetracycline was equal to or higher than 64 micrograms/ml in 19 strains (76%), whereas for 5 strains the MIC was 1 microgram/ml and for 1 strain 2 micrograms/ml.

Gene, 1993 Nov 30, 134(1), 1 - 6
Sequence analysis of four new heat-shock genes constituting the hslTS/ibpAB and hslVU operons in Escherichia coli; Chuang SE et al.; Sequences of four new heat-shock (HS) genes of Escherichia coli organized into two operons were determined . The operon at 83 min specifies two proteins of 15.8 kDa (HslT) and 16.1 kDa (HslS), which are identical to IbpA and IbpB, respectively . Expression of mRNA from a sigma 32-dependent promoter of the hslTS/ibpAB operon is stimulated 30-75-fold upon temperature upshift . The transcription start point (tsp) is located at a G, 96 bp upstream from the AUG start codon of hslT/ibpA . The deduced amino acid sequences of HslT/IbpA and HslS/IbpB are 48% identical to each other and were found to be remotely related to the chloroplast low-molecular-weight HS protein, which is highly conserved among plants . The second hs operon is much less actively stimulated by temperature upshift, although it has a hs promoter that perfectly matches the consensus of promoters recognized by sigma 32 . Located at 88.9 min, the hslVU operon specifies proteins of 19.1 kDa (HslV) and 49.6 kDa (HslU) . Multiple tsp were found in this operon . HslV is remotely related to the eukaryotic proteasome proteins, and HslU is very similar to a Pasteurella haemolytica protein of unknown function . Both HslU and the P . haemolytica protein share a ATP/GTP-binding motif near their N-termini . The two operons described here are transcribed counterclockwise on the standard genetic map.

Infect Immun, 1993 Nov, 61(11), 4909 - 14
Isolation and nucleotide sequence of the gene encoding cytotoxic necrotizing factor 1 of Escherichia coli; Falbo V et al.; Cytotoxic necrotizing factors (CNFs) are dermonecrotic protein toxins produced by human and animal clinical isolates of Escherichia coli . In this study, the CNF1 determinant was isolated and sequenced, showing that expression of biologically active toxin is governed by a unique open reading frame encoding a protein of 1,014 amino acids with a predicted molecular mass of 113.7 kDa . Nucleotide and protein data base searches showed significant homology between CNF1 and the dermonecrotic toxin of Pasteurella multocida . In particular, the two toxins were found to share a hydrophobic region of about 220 amino acids which is a potential membrane-spanning domain.

Infect Immun, 1993 Nov, 61(11), 4785 - 92
Virulence of capsulated and noncapsulated isolates of Pasteurella multocida and their adherence to porcine respiratory tract cells and mucus; Jacques M et al.; The virulence and the adherence to porcine respiratory tract cells and mucus of three toxigenic, capsular type D Pasteurella multocida isolates and their noncapsulated variants were evaluated in the present study . Loss of capsule by P . multocida, verified by transmission electron microscopy after polycationic ferritin labeling, was associated with a massive reduction in virulence of the organisms in mice . Specific-pathogen-free piglets inoculated intranasally with one of the capsulated isolates or its noncapsulated variant developed turbinate lesions characterized by bone resorption and by an inflammation of the mucosa associated with hyperplasia and squamous metaplasia of the epithelium . Infection with the capsulated isolate led to more severe lesions and atrophy of turbinates . The interactions of these P . multocida isolates with porcine respiratory tract cells and mucus were studied in vitro . The presence of capsule resulted in a decrease in binding of respiratory tract mucus were studied in vitro . The presence of capsule resulted in a decrease in binding of respiratory tract mucus to P . multocida isolates as determined by a dot blot assay . The presence of capsule also resulted in a significant decrease in adherence to porcine tracheal rings maintained in culture . The capsule seemed to mask outer membrane components which are involved in adherence . One of these components might be lipopolysaccharide since purified lipopolysaccharide bound respiratory tract mucus and blocked adherence of this microorganism to porcine tracheal rings . Our data indicate that capsular material does not seem to be involved in adherence of P . multocida to respiratory tract cells and mucus, but capsulated isolates are more virulent in mice and also in piglets.

Infect Immun, 1993 Nov, 61(11), 4669 - 74
Characterization of neuraminidases produced by various serotypes of Pasteurella haemolytica; Straus DC et al.; Neuraminidases produced by 16 strains of Pasteurella haemolytica (serotypes 1 to 16) were characterized by molecular weight, antigenic identity, and substrate specificity . After growth in a chemically defined medium, stage I (lyophilized) culture supernatants were assayed for activity with N-acetylneuramin lactose, human alpha-1-acid glycoprotein, fetuin, colominic acid, and bovine submaxillary mucin . Neuraminidase produced by P . haemolytica serotype A1 (Ph A1) was purified by a combination of salt fractionation, ion-exchange chromatography on DEAE-Sephacel, and gel filtration on Sephadex G-200 . Purified Ph A1 neuraminidase was used to immunize rabbits, and the resultant antiserum reduced the activity of Ph A1 neuraminidase by 46% . This antiserum also reduced the activity of neuraminidase produced by the other serotypes by between 15 and 66% . Molecular weight estimates of the neuraminidases produced by the various serotypes were obtained by gel filtration chromatography on Sephadex G-200 . Fifteen of the 16 serotypes examined produced a neuraminidase with a molecular weight of approximately 150,000 to 200,000 . One serotype (serotype 11) produced no material with neuraminidase activity . In addition, all 15 high-molecular-weight neuraminidases showed similar substrate specificities . That is, they were all most active against N-acetylneuramin lactose and least active against bovine submaxillary mucin . On the basis of these results, it appears that the high-molecular-weight neuraminidases produced by the different P . haemolytica serotypes are quite similar.

Enferm Infecc Microbiol Clin, 1993 Nov, 11(9), 482 - 6
{Bite wound infections: study of 22 hospitalized patients}; Gasser I et al.; BACKGROUND: To study the characteristics of bite wounds with unfavorable evolution, developing infectious complications that requires hospital admission . METHODS: The data from 22 patients admitted to the Ciudad Sanitaria Vall d'Hebron hospital for the above mentioned reason over the last 5 years were reviewed . RESULTS: The patients (8 males, 8 females and 6 children) were bitten by 10 dogs, 6 cats, and 6 men with predominance of the wound site being in the upper limb (10) followed by the lower limbs (6), head or face (5) and exceptionally on the breast (1) . The most frequent clinical manifestation was abscess and/or cellulitis (13) and adenopathies or lymphangitis (4); 5 patients presented osteoarticular involvement including 3 bone fractures due to human aggression . With regard to the etiology of infection, the common bucal flora bacteria were isolated in all the cases; Pasteurella multocida in 15/16 animal bites, Eikenella corrodens associated to streptococcus in 5/6 human bites, Fusobacterium spp . (5), Bacteroides spp . (3) and Peptococcus sp . (1) . The most frequently administered antibiotics were gentamycin (15), penicillin (13), cloxacillin (5) and clindamycin (4) . The evolution was favorable, although slow in many cases, with sequelae in 3 patients . CONCLUSIONS: It is very difficult to foresee in which cases infectious complications will develop in bite wounds . According to the authors' experience, in the case of deep wounds the bacteria implied come from the mouth of the aggressor . Careful cleansing, rapid administration of an adequate antibiotic and clinical control being the most recommendable procedure.

Plasmid, 1993 Nov, 30(3), 268 - 73
Construction of a shuttle vector for use between Pasteurella multocida and Escherichia coli; Bills MM et al.; The isolation of a small plasmid from Pasteurella multocida has enabled the construction of a shuttle vector for use between P . multocida and Escherichia coli . The vector pBAC64 contains the origin of replication from P . multocida, an antibiotic resistance gene which functions in P . multocida, and the E . coli vector pUC18 . The presence of the pUC18 multiple cloning site together with the lacZ' gene provides a screening method and allows cloning and manipulation in E . coli as well as cloning in P . multocida.

Res Vet Sci, 1993 Nov, 55(3), 317 - 25
Purification and quantitative measurement of bovine serum amyloid-A; Horadagoda A et al.; Bovine serum amyloid-A (b-SAA) was purified from a pool of acute phase serum using hydrophobic interaction chromatography and gel filtration . Serum was applied at a low salt concentration to a phenyl-sepharose column and SAA was eluted with a gradient of 0 to 6 M guanidine-HCl . Fractions containing SAA were pooled, concentrated and further purified by gel filtration on Superose-12 . The concentration of SAA in bovine serum was quantified by an indirect ELISA using rabbit anti-human SAA and horseradish peroxidase conjugated donkey anti-rabbit IgG . Dilutions of an acute phase bovine serum sample were used as working standards . The SAA concentration of this standard was determined by comparison with purified b-SAA on SDS-polyacrylamide gel electrophoresis followed by densitometry at 590 nm . The assay detection limit was 3 micrograms ml-1; the intra-assay coefficient of variation was 4 per cent and interassay coefficients of variation were 5.5 per cent and 7.2 per cent at 66 and 178 micrograms ml-1 SAA, respectively . In calves experimentally infected with Pasteurella haemolytica type A1 the ELISA was able to detect a 10-fold increase of SAA within 24 hours of inoculation.

Kansenshogaku Zasshi, 1993 Nov, 67(11), 1041 - 4
{Studies on respiratory infection with Pasteurella multocida}; Arashima Y et al.; A questionnaire style survey on Pasteurella multocida infection was conducted at on 528 laboratories of hospitals . The questionnaire included the detection of Pasteurella multocida from sputum, the age distribution, underlying disease etc . of the patients . In Japan, infectious diseases caused by Pasteurella multocida were found in 179 cases in 67 (26.0%) of the 258 hospitals . The incidence of the infections tends to increase, 18 cases in 1989, 25 cases in 1990, 18 cases in 1991 (Jan.-Jul.) . These 179 cases were broken down into 44 males (from 1 year old to 85 years old, mean 58.9 years old), 45 females (from 18 years old to 85 years old, mean 60.2 years old), and 90 patients whose sex was unknown . Underlying diseases were recognized in 72 cases . Diseases related to the respiratory organ were 61 (84.7%) cases . The cases of hemo-sputum had to be differentiated from malignant tumors Recurrent Pasteuralla multocida infections were recognized in 4 males and 4 females . And the 4 cases had been handling animals . There were only 26 cases who had contact with animals, the others were unknown . This result suggested that contact with animals may cause these infections . Recently, the detection of Pasteurella multocida from the sputum tends to increase . The elderly patient with chronic pulmonary disease, who is handling animals must be educated on hygiene and zoonosis.

Br Vet J, 1993 Nov-Dec, 149(6), 561 - 70
Pasteurella haemolytica serotypes isolated in Northern Ireland during 1989-1991; Ball HJ et al.; Pasteurella haemolytica isolates from clinical veterinary samples were serotyped over a three year period to determine the distribution of strains in Northern Ireland, to compare the results with the recorded distribution in the remainder of Great Britain and to extend information on the geographical prevalence of the serotypes . Four hundred and nine typable and 91 untypable strains were isolated from 490 animal cases . From 127 typable cattle isolates, the commonest serotype was A1, most frequently associated with calf pneumonia . Serotypes A2, T4, T10 and T15 were the commonest sheep isolates, A2 being most frequently associated with lamb pneumonia and the T-biovars more prevalent in adult pneumonia and lamb septicaemia . The isolates from approximately half the cases were evaluated in terms of the results obtained from gross and histopathological post mortem examination . Approximately two-thirds of these isolates were associated with significant pathology, including a small number of serotypes which are not included in the sheep vaccines currently used in Great Britain.

Br Vet J, 1993 Nov-Dec, 149(6), 549 - 60
Morbidity and mortality in a large Irish feedlot; microbiological and serological findings in cattle with acute respiratory disease; Healy AM et al.; A survey of morbidity and mortality was undertaken in a slatted unit containing 6399 beef cattle over a 6 month period . The mortality rate was 0.78% and the morbidity rate was 12.7% . The interval from arrival to morbidity was 25.5 days +/- 0.9 . Significantly more sick cattle were identified on either Mondays or Tuesdays than were seen on Saturdays or Sundays (P < 0.005) . The mean interval to mortality (all diseases) was 48.5 days +/- 7.4 . Respiratory disease was the most frequently recorded disease and was responsible for 62% of morbidity and 58% of mortality . Samples from 133 cattle, taken as they presented with acute onset respiratory disease, contained antibodies to Bovine Herpes Virus -1(BHV-1) (14.3%), Bovine Viral Diarrhoea Virus (BVDV) (78%), Bovine Respiratory Syncytial Virus (BRSV) (94%) and Parainfluenza type 1 Virus (P13V) (99%) . When the same cattle were resampled 4 to 6 weeks later antibodies were found for BHV-1 (93.2%), BVDV (99.2%), BRSV (100%) and P13V (100%) . Eleven of 27 bronchoalveolar lavage samples taken from the above group of cattle exhibited positive fluorescence for BHV-1 but all 27 samples were negative for BVDV, BRSV and P13V . Pasteurella multocida was isolated from eight of the 27 lavage samples and Pasteurella haemolytica from one sample.

Mikrobiol Z, 1993 Nov-Dec, 55(6), 50 - 6
{The antagonistic activity of the bacterial flora in feed in relation to the causative agent of diarrhea in calves}; Kharchenko SN et al.; Antagonistic activity of 264 strains (6 species) of aerobic bacilli as well as 45 strains (3 species) of bacteria from genera Chromobacterium and Pseudomonas has been studied . They were isolated from the fodder at the farms of Ukraine five regions in 1986-1992 in respect to 128 strains (17 species) of genera Escherichia, Proteus, Salmonella, Shigella and Pasteurella, isolated by the authors from feces of calf with symptoms of diarrhea . High antagonistic activity of Bacillus polymyxa 168 (zone of growth inhibition of test cultures of 10-20 mm) in respect of diarrhea agents was established . The per os application of B . polymyxa in a dose of 200-250 ml of water suspension containing 100 bill . cells accelerated the process of recovery in the calves with symptoms of diarrhea as well as increased the weight increase in newborn calves.

Intern Med, 1993 Nov, 32(11), 872 - 4
Pasteurella ureae endocarditis; Yamamoto K et al.; Pasteurella ureae is found in the normal human respiratory flora . We encountered a case of endocarditis caused by Pasteurella ureae . The patient was a 59-year-old man with a history of Staphylococcus aureus endocarditis . After treatment with antibiotics, blood cultures became negative, and the patient recovered completely . The incidence of endocarditis due to this organism is very rare; this case is the second clinically diagnosed case report.

Biol Pharm Bull, 1993 Nov, 16(11), 1065 - 8
Reduction of endotoxin contamination of various crude vaccine materials by gram-negative bacteria using aminated poly(gamma-methyl L-glutamate) spherical particles; Sakata M et al.; We describe a method for the removal of endotoxins from various crude antigen solutions originating from gram-negative bacteria using aminated poly(gamma-methyl L-glutamate) (PMLG) spherical particles . The aminated PMLG adsorbents showed high affinity for various purified endotoxins at an ionic strength of mu = 0.1 . The endotoxin-adsorbing capacity of the adsorbent increased with increase in the amino-group content of the adsorbent . The adsorbent (3.2 meq/g amino-group content) showed the highest affinity for endotoxin at ionic strengths ranging from mu = 0.025-0.8 . The adsorption of Bordetella pertussis antigen to the adsorbent decreased with increasing amino-group content of the adsorbent at an ionic strength of mu = 0.2 . The adsorption of B . bronchiseptica protein to the adsorbent increased with increasing amino-group content of the adsorbent, but decreased with increasing ionic strength . The adsorbent (3.2 meq/g of amino-group content) selectively reduced endotoxin in crude antigen solutions originating from gram-negative bacteria, B . pertussis, B . bronchiseptica and Pasteurella multocida, even at a high ionic strength (mu = 0.2-0.4) without affecting the recovery of the protective antigens.

Cornell Vet, 1993 Oct, 83(4), 303 - 9
Combined enzyme and antimicrobial susceptibility profiles of caprine Pasteurella haemolytica serovar 2 respiratory tract isolates; Scanlan CM et al.; The present investigation describes a novel method of demonstrating strain diversity among Pasteurella haemolytica biovar A, serovar 2 (PhA2) nasal turbinate isolates from a flock of 32 experimental goats during a naturally occurring outbreak of pasteurellosis . After a 21 day conditioning period in a feedyard, 51 PhA2 isolates from 27 culture-positive goats were identified including 1 on day 22, 14 on day 25, 21 on day 39, and 15 on day 66 . Each PhA2 isolate was evaluated for its enzyme activity against 19 substrates with a commercial semiquantitative enzyme system and for its antimicrobial susceptibility with 12 drugs, resulting in 7 different enzyme profiles and 8 different antimicrobial susceptibility profiles . A total of 14 combined enzyme and antimicrobial susceptibility profiles were produced . The same PhA2 strain was isolated from only 4 of the 12 goats with 2 PhA2 isolations, while the same PhA2 strain was isolated from only 1 of the 6 goats with 3 PhA2 isolations . The data from this investigation demonstrated that the PhA2 upper respiratory tract flora from goats is highly heterologous.

J Comp Pathol, 1993 Oct, 109(3), 303 - 7
The effects of tick-borne fever on the susceptibility of ovine polymorphonuclear cells to P . haemolytica cytotoxin; Woldehiwet Z et al.; Tick-borne fever predisposes sheep to secondary infections by Pasteurella haemolytica . Polymorphonuclear (PMN) cells obtained from sheep 2 to 7 days after the onset of parasitaemia due to Cytoecetes phagocytophila, the causative agent of tick-borne fever, were significantly more susceptible in vitro to the cytotoxin of P . haemolytica than PMN obtained from age-matched control sheep.

Vet Microbiol, 1993 Oct, 37(1-2), 31 - 43
Molecular analysis of a cryptic plasmid isolated from avian strains of Pasteurella multocida; Price SB et al.; Twelve small plasmids isolated from avian strains of Pasteurella multocida were examined by restriction enzyme mapping, cross-hybridization, and minicell analysis . These plasmids contained sites for several commonly used restriction enzymes and ranged in size from 3.4 to 3.8 kilobases . Restriction enzyme maps of the 12 plasmids were similar and divided the plasmids into 3 families, designated pFS1, pFS2, and pFS4 . Restriction fragments of pFS1 DNA isolated from strain X-73 were used to probe AvaI/HindIII/EcoRV digests of pFS2 and pFS4 DNA . The results of these hybridization experiments demonstrated that the plasmids found in all three families shared extensive regions of homology and may have originated from a common ancestor . Escherichia coli minicells containing recombinant plasmid constructs bearing fragments of pFS1 expressed two pFS1-specific peptides, 12.5 and 28 kilodaltons in size, suggesting that some P . multocida plasmid-encoded proteins can be expressed in E . coli . These results indicate that pFS may be useful as a genetic tool for moving DNA into and out of P . multocida, since it is small, contains common restriction sites, and encodes at least two genes that are recognized and expressed in E . coli.

J Vet Diagn Invest, 1993 Oct, 5(4), 555 - 9
Systemic Pasteurella haemolytica infection as a rare sequel to avirulent live Pasteurella haemolytica vaccination in cattle; Zeman D et al.; Eleven cases of systemic Pasteurella haemolytica infection in cattle were identified from routine diagnostic laboratory submissions during the falls of 1988, 1989, and 1991 . All cases came with a history of recent vaccination with an avirulent live culture P . haemolytica product . Nine of 11 cases involved cattle vaccinated between 2 and 18 days previously with this product . Ten of 11 cases involved 182-227-kg beef calves that were vaccinated between September and November during routine processing for entry into feedlots . The morbidity and mortality was generally low . The major pathologic findings included meningitis, injection site abscessation and/or cellulitis, and polyarthritis . Systemic infection was indicated in all cases by the isolation of P . haemolytica from 2 or more organs or distinct anatomical sites . In 6 cases, the vaccine injection site was cultured, and in all 6 cases, P . haemolytica was isolated . Three separate P . haemolytica isolates from 2 cases were further studied by restriction enzyme analysis (REA) . These isolates were from tissues with suppurative inflammation, including the brain, joint, and injection site . The REA patterns of each of these 3 isolates were identical to the REA pattern of the vaccine masterseed, which strongly suggested that the organisms causing systemic infection were the same as the organism used to produce the vaccine . Because the overall incidence was quite low, other factors, such as stress, probably played a major role in the expression of this syndrome.

J Wildl Dis, 1993 Oct, 29(4), 582 - 6
Lead concentrations in liver and kidneys of snow geese during an avian cholera epizootic in California; Gordus AG; During an avian cholera epornitic, between December 1982 and January 1983, 58 dead, 23 sick, and 106 hunter-killed lesser snow geese (Chen caerulescens caerulescens) were collected at Delevan National Wildlife Refuge, Colusa County, California, USA . Fifty-one of the dead and sick geese were infected with Pasteurella multocida . Lead concentrations in the livers ranged from < 1 to 253 parts per million (ppm) (dry weight) . Lead concentrations in the kidneys ranged from < 1 ppm to 547 ppm (dry weight) . Snow geese with > 30 ppm lead, considered diagnostic of acute lead poisoning, had significantly (P < 0.05) lower heart weights and a smaller band of heart fat, compared to geese with tissue lead concentrations of < 30 ppm . Tissue lead concentrations in geese dying from avian cholera generally were lower than concentrations in hunter-killed geese, but the differences were not significant for either kidney (P = 0.08) or liver (P = 0.30) tissue.

Berl Munch Tierarztl Wochenschr, 1993 Oct, 106(10), 333 - 6
{In vitro adherence of Pasteurella haemolytica to tracheal mucins and a tracheal epithelial cell preparation from cattle}; Botcher L et al.; Outer membrane protein preparations (OMP) of 11 Pasteurella haemolytica field isolates of the serovars A1, A2, A11 and non-typable strains from cattle were extracted by N-lauryl-sarcosine sodium salt . Capsular extracts were prepared by heat treatment . Both preparations and whole cell suspensions bound to a preparation of an epithelial cell wall fraction of trachea and to tracheal mucus of cattle . Binding was demonstrated by enzyme linked immunosorbent assay (ELISA) . Distinct high binding values were shown by the OMP and the capsular extracts of the serovar A1-strains.

Berl Munch Tierarztl Wochenschr, 1993 Oct, 106(10), 330 - 3
{Pneumonia diagnosis in living swine using lung lavage}; Ganter M et al.; In pigs coming from fattening units with tenacious pneumonitis problems the attempt was made to find an etiological diagnosis in living pigs by bronchoalveolar lavage (BAL) and serological examinations on antibodies against Mycoplasma hyopneumonia, Actinobacillus pleuropneumoniae and Influenza-A-virus (serotypes H1N1 and H3N2) . In some cases the results of the bacteriological examinations of the BAL were compared with the post mortem findings . Both methods yield similar results . Mycoplasma hyopneumonia could neither be cultured from the BAL nor indicated the results of the serological examinations infections with M . hyopneumoniae . Pasteurella multocida was isolated from 73% of the BAL-samples . Actinobacillus pleuropneumoniae was found in 4 from 90 samples . Positive antibody titers against Influenza-virus in most of the sera indicate that the clinical course of Influenza is changing from an acute epidemic to an enzootic disease in fattening units.

Berl Munch Tierarztl Wochenschr, 1993 Oct, 106(10), 328 - 30
{The detection of Pasteurella multocida toxin using a commercially available ELISA}; Weber A et al.; 126 Pasteurella (P.) multocida strains isolated from nasal swabs of pigs were investigated for producing toxin, comparing tissue culture (EBL cells) and DAKO ELISA-PMT-Kit . With help of ELISA 13 toxigenic P . multocida strains were identified, with help of tissue culture 12 strains producing toxin . In 14 of 219 examined nasal swab bacteria cultures of pigs toxin of P . multocida was detected by the commercially available ELISA . Because of an abundant mixed bacteria culture in 7 of these 14 swabs suspected colonies of P . multocida could not be isolated.

Am J Vet Res, 1993 Oct, 54(10), 1637 - 47
Efficacy of Pasteurella haemolytica subunit antigens in a goat model of pasteurellosis; Purdy CW et al.; The effectiveness of Pasteurella haemolytica biovar A, serovar 1 (Ph1) subunit vaccines was tested in goats, using challenge exposure by transthoracic injection . Twenty-two weanling male Spanish goats were randomly allotted to 4 groups . Six goats were given 2 transthoracic injections into the lung 18 days apart with live Ph1 impregnated in agar beads (positive controls) . Six goats were not given injections (negative controls) . Five goats were given 2 transthoracic injections into the lung 18 days apart with 4.6 mg of cytotoxin in agar beads . The remaining 5 goats were given 2 IM injections, 18 days apart, into the thigh with 4.6 mg of cytotoxin emulsified in incomplete Freund's adjuvant . Twenty-four days after the second injection, all goats were challenge-exposed to live Ph1 by transthoracic injection into the lung, and 4 days later, all goats were euthanatized and necropsied . Serum neutralizing anticytotoxin titer was measured throughout the experiment . Mean volume of consolidated lung tissue was 0.38 cm3 for the positive control group, 32 cm3 for the negative control group; 19 cm3 for the cytotoxin-lung group; and 88 cm3 for the cytotoxin-adjuvant-IM group . Only the positive control group was protected from Ph1 challenge exposure . The Ph1 cytotoxin subunit vaccine alone appeared to be ineffective, and the anticytotoxin titer was not correlated with protection . In a separate trial, 32 weanling male Spanish goats were randomly allotted to 5 groups . Each was given 2 transthoracic injections into the lung 22 days apart . Six goats were given Ph1 cytotoxin impregnated into agar beads; 6 were given Ph1 lipopolysaccharide impregnated in agar beads; 6 were given Ph1 capsule impregnated in agar beads . Six goats were given agar beads only (negative controls), and 6 were given live Ph1 impregnated into agar beads (positive controls) . Twenty days after the second injection, all goats were challenge-exposed to live Ph1 by transthoracic injection into the lung, and 4 days later, all goats were euthanatized and necropsied . Mean volume of consolidated lung tissue was 0.14 cm3 for the positive control group, 7.59 cm3 for the negative control group, 11.21 cm3 for the cytotoxin group, 10.19 cm3 for the lipopolysaccharide group, and 1.6 cm3 for the capsule group . Again, only injection of live Ph1 (positive controls) induced solid protection; however, the capsule subunit vaccine induced partial protection against challenge exposure in this trial . Lipopolysaccharide and cytotoxin subunit vaccines were ineffective in protecting goats against challenge exposure with live Ph1.

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