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 memb