Pasteurella

Pathology, 1993 Oct, 25(4), 379 - 84
Dog-associated bacterial infections in humans: isolates submitted to an Australian reference laboratory, 1981-1992; Peel MM; Over the period 1981-92, 32 bacterial isolates were referred to the Microbiological Diagnostic Unit from infected dog-bite wounds and 10 isolates were submitted from blood cultures after dog bites or close contact with dogs . The isolates from the bite wounds were identified, or confirmed, as Pasteurella multocida (11 isolates), Pasteurella dagmatis (3), CDC group M-5 (9), CDC group EF-4a (8) and Streptococcus anginosus (1) . Five of the 9 patients from whom CDC group M-5 was cultured had mixed infections: 2 with P . multocida one of which also had Bacteroides sp., one with P . dagmatis, one with CDC group EF-4a and another with Bacteroides sp . Nine of the 10 blood isolates were identified as Capnocytophaga canimorsus . The remaining one of Streptobacillus moniliformis, which is typically associated with rat-bite fever, was the result of a bite from a breed of dog (greyhound) that eats rodents . Clinical notes are provided for infections caused by the more unusual bacterial isolates and their laboratory identification is described.

Avian Dis, 1993 Oct-Dec, 37(4), 1121 - 9
Facial cellulitis associated with fowl cholera in commercial turkeys; Jeffrey JS et al.; Severe cephalic swelling and facial cellulitis in turkeys associated with fowl cholera were present in seven accessions submitted to two laboratories in a 2-year period . Flocks ranged in age from 6 to 18 weeks and included both toms and hens . Interestingly, turkeys with facial cellulitis had no gross internal lesions of fowl cholera, whereas birds with gross lung, liver, and air-sac lesions did not have swollen heads . Histologically, the facial cellulitis was characterized by extensive fibrinonecrotic inflammation of the deep dermis with heterophilic perivasculitis and thrombosis . Additional characterization of Pasteurella multocida isolates from these cases was conducted retrospectively from lyophilized cultures . Serogrouping, serotyping, and an enzyme-linked immunosorbent assay (ELISA) for dermonecrotic factor were performed . All isolates were serogroup A or unencapsulated . Serotype 1 was the most prevalent serotype isolated in association with facial cellulitis . ELISA results for dermonecrotic toxin were inconclusive.

Avian Dis, 1993 Oct-Dec, 37(4), 1074 - 9
Cross-protection studies with Pasteurella multocida bacterins prepared from bacteria propagated in iron-depleted medium; Glisson JR et al.; Strains X-73 (serotype 1) and P-1059 (serotype 3) of Pasteurella multocida, avian origin, expressed additional membrane proteins (MPs) when grown in brain-heart infusion (BHI) broth containing the iron chelator dipyridyl and when grown in BHI broth treated with the iron chelator Chelex 100 . These additional MPs were not detected when both strains were grown in BHI broth . Chickens and turkeys were vaccinated twice with inactivated oil-emulsion vaccines containing bacterial cells expressing these MPs or with vaccines containing bacterial cells grown in BHI broth . Two weeks after the final vaccination, all birds were challenged to determine whether bacterins made from P . multocida that had been propagated in conditions of iron deprivation would induce heterologous serotype immunity . The bacterins produced in medium low in iron did not consistently induce significant protection against heterologous challenge.

Avian Dis, 1993 Oct-Dec, 37(4), 1071 - 3
Pasteurella multocida virulence factors: selection of fowl cholera-inducing and non-inducing strains; Rhoades KR et al.; Relatively little information is available on Pasteurella multocida virulence factors involved in producing fowl cholera . Because of the complex nature of bacterial pathogenesis, the recommended approach for ascertaining these factors is to compare biological attributes of high- and low-virulence strains . To permit use of this approach for fowl cholera, P . multocida strains of high and low virulence were identified . Turkey poults were exposed intrapharyngeally and intravenously (IV) to two antigenically and biochemically similar strains . Based on mortality, strain P-1059 was highly virulent and strain P-1062 was avirulent . Microbiological examination indicated that only the virulent strain infected the pharyngeal mucosa of intrapharyngeally exposed poults and survived and multiplied in IV-exposed poults . These findings indicate strain differences in those virulence factors concerned with the colonization and multiplication stages of disease development.

Rev Latinoam Microbiol, 1993 Oct-Dec, 35(4), 361 - 9
Adherence of Pasteurella multocida to rabbit respiratory epithelial cells in vitro; Bonilla-Ruz LF et al.; Adult clinically healthy New Zealand rabbits were sampled bacteriologically to detect carriers and non-carriers of Pasteurella multocida . Both groups of rabbits were killed separately to obtain samples of nasal, buccal, pharyngeal and tracheal epithelial cells . The cells were tested for adherence in vitro to 18 isolates of P . multocida from healthy and sick rabbits, from ovine, bovine, cat and swine . The number of bacteria adhered per cell up to 25 cells per preparation were registered . Analysis of variance was used to interpret the significance of results . Adherence of P . multocida was significantly higher to carrier rabbit cells than to non-carrier rabbit cells . Bacterial isolates from rabbits were more adherent to rabbit cells than to isolates from other species . The frequency was higher to buccal and pharyngeal cells than to nasal and tracheal cells . Isolates from healthy animals adhered better to rabbit cells than isolates from sick animals, except the isolates from sick animals which adhered better to nasal cells of non carriers than did isolates from healthy rabbits.

Kansenshogaku Zasshi, 1993 Sep, 67(9), 791 - 4
{Current status of Pasteurella multocida infection in Japan}; Arashima Y et al.; Recently, the case reports of Pasteurella multocida infection has been increasing in Japan . In 1989, the Japanese Government, Veterinary Sanitation Division, Ministry of Health and Welfare officially communicated this infection as a zoonosis to related institutions . The current status of Pasteurella multocida infection is not well known in Japan . Because of this, a nation wide questionnaire survey on Pasteurella multocida was conducted to clarify the status . A questionnaire was sent to 380 laboratories of the hospitals, and 258 (67.9%) replied . An infectious disease caused by Pasteurella multocida was found in Japan in 369 cases in 115 (44.6%) of 258 hospitals, or an average of 3.2 cases per hospital . The 369 cases were broken down into 123 males (from 1 month old to 87 years old), 118 females (from 7 months old to 88 years old), and 128 patients whose sex was unknown . The incidence of the infections tends to increase year by year . This incidence is higher than our expectation . It is considered that the contact with pets will in increase the infection with this agents . The organism was isolated in as many as nineteen different body specimens, including the appendix and urine, which in Japan has not been reported as organs harboring this organism . Some of the nineteen cases were severely infected . This organism was isolated most often from the sputum (48.5%) . Pus was the next most common site (27.1%) . This order was reversed in the U.K . and the U.S. . Possible explanations for the reversal are given below.(ABSTRACT TRUNCATED AT 250 WORDS)

J Am Vet Med Assoc, 1993 Sep 1, 203(5), 667 - 9
Ovarian abscesses and pyometra in a domestic rabbit; Johnson JH et al.; Infection with Pasteurella multocida can induce pyoendometritis, pyosalpingitis, and ovarian abscesses in rabbits . The likelihood of rabbits developing clinical pasteurellosis is predisposed by factors such as buildup of ammonia fumes, ambient temperature changes and drafts, reproduction, older age, existence of carriers, and poor sanitation . Traditionally, prevention of pasteurellosis in rabbits has focused on identifying suspected carriers and culling those animals from the rabbitry.

Infect Immun, 1993 Sep, 61(9), 3942 - 51
Expression of the Pasteurella haemolytica leukotoxin is inhibited by a locus that encodes an ATP-binding cassette homolog; Highlander SK et al.; Multicopy and single-copy chromosomal fusions between the Pasteurella haemolytica leukotoxin regulatory region and the Escherichia coli beta-galactosidase gene have been constructed . These fusions were used as reporters to identify and isolate regulators of leukotoxin expression from a P . haemolytica cosmid library . A cosmid clone, which inhibited leukotoxin expression from multicopy and single-copy protein fusions, was isolated and found to contain the complete leukotoxin gene cluster plus additional upstream sequences . The locus responsible for inhibition of expression from leukotoxin-beta-galactosidase fusions was mapped within these upstream sequences, by transposon mutagenesis with Tn5, and its DNA sequence was determined . The inhibitory activity was found to be associated with a predicted 440-amino-acid reading frame (lapA) that lies within a four-gene arginine transport locus . LapA is predicted to be the nucleotide-binding component of this transport system and shares homology with the Clp family of proteases.

Res Vet Sci, 1993 Sep, 55(2), 209 - 14
Influence of Newcastle disease virus on the severity of Pasteurella anatipestifer infection in turkeys; Charles SD et al.; This study was designed to examine whether vaccine or virulent strains of Newcastle disease virus (NDV) would potentiate the disease caused by Pasteurella anatipestifer infection in turkeys . The studies were conducted in turkeys of two age groups . There were three experiments . In two experiments four-week-old turkeys were exposed either to vaccine or virulent strains of NDV after experimental P anatipestifer infection . In the third experiment 14-week-old turkeys were first exposed to virulent NDV and superimposed with P anatipestifer infection . In experiment 1, one bird died where P anatipestifer was given in combination with the vaccine strain of NDV . However, there was no difference in the clinical signs, gross lesions and histopathology compared with turkeys given P anatipestifer alone . In experiment 3 where turkeys received a virulent strain of NDV in combination with P anatipestifer, birds became dyspnoeic and showed signs of illness . There was a difference in the course of the disease, gross lesions and histopathology when compared with turkeys that received P anatipestifer only.

Dtsch Tierarztl Wochenschr, 1993 Sep, 100(9), 355 - 9
{The effect of a Bordetella live vaccine on the occurrence and manifestation of atrophic rhinitis suum and the aerogenous infection burden in field strains of the agent}; Ehser U et al.; A field study was carried out in a large scale unit for swine breeding and fattening with the object to influence the high morbidity rate of Atrophic Rhinitis and pneumonia with the help of a Bordetella live vaccine . The results show that it is possible to decrease the infectious pressure by B . bronchiseptica and to reduce the pathomorphological signs of Atrophic Rhinitis in consequence of the application of the live vaccine . The pathologic-anatomical investigations of nasal turbinates in immunized slaughtered fattening pigs show a significant lower morbidity concerning Atrophic Rhinitis and a higher percentage of pigs without changes at conchae nasales and septum nasi . We find also a lower contamination of the air with B . bronchiseptica field strains during vaccine application . The results also explain that a high infectious pressure by B . bronchiseptica and the possibility of communication between unvaccinated and vaccinated groups of pigs counteract a better efficiency of the vaccine . The decrease of the morbidity rate of Atrophic Rhinitis appears so much more important because toxicogenic Pasteurella multocida strains were isolated from nasal swabs of vaccinated pigs during the investigations . But these strains influenced the Atrophic Rhinitis frequency only accidentally . All results as a whole point out that in pig houses with a high animal density one has to pay more attention to virulent B . bronchiseptica strains than it was been done till now.

J Clin Microbiol, 1993 Sep, 31(9), 2303 - 8
Restriction endonuclease analysis and ribotyping differentiate Pasteurella haemolytica serotype A1 isolates from cattle within a feedlot; Murphy GL et al.; Pasteurella haemolytica serotype A1 isolates were collected from cattle within a feedlot during an outbreak of bovine respiratory disease . Genetic heterogeneity among the isolates was examined by restriction endonuclease analysis (REA), ribotyping, and analysis of plasmid content . The susceptibilities of isolates to several antibiotics were also examined . Five different REA patterns and three different ribotypes were observed among the isolates . Fifty percent of the isolates had an identical REA type, ribotype, and plasmid profile . Examination of the plasmid content of the isolates revealed that most (73%) carry a single plasmid which encodes beta-lactamase, 13.5% carry two plasmids, and 13.5% carry no plasmid . The data reveal the presence of genetic differences among isolates of P . haemolytica A1, associated with shipping fever pneumonia within a closed feedlot, and suggest that a combination of REA, ribotyping, plasmid analysis, and antibiotic susceptibility determination will be useful in analyzing the molecular epidemiology of this disease.

FEMS Immunol Med Microbiol, 1993 Aug, 7(2), 105 - 10
An experimental anti-idiotype vaccine mimicking lipopolysaccharide gives protection against Pasteurella multocida type A infection in mice; Sutherland AD et al.; An anti-idiotype strategy was employed which showed that polyclonal anti-idiotype antibodies could be produced which could mimic a linear Pasteurella multocida lipopolysaccharide (LPS) molecule . These antibodies when used as vaccine antigens, induced antibodies which recognised LPS and imparted acquired protection upon syngeneic vaccinates challenged with homologous organisms.

J Vet Med Sci, 1993 Aug, 55(4), 617 - 22
Interaction between immunity to Bordetella bronchiseptica and infection of pig herds by Bordetella bronchiseptica and Pasteurella multocida; Elias B et al.; The dynamics of toxigenic Bordetella bronchiseptica and Pasteurella multocida infection and the B . bronchiseptica specific antibody content of the blood and nasal secretion were studied in three Hungarian and three Dutch pig herds . In both countries, the studies involved young sows that had farrowed once or twice (YS), old sows that had farrowed more than four times (OS), and their piglets . The results indicate that Dutch sows are characterized by a lower prevalence of B . bronchiseptica and P . multocida infection than Hungarian sows . In Dutch sows and in their piglets, the rate of P . multocida infection was higher than that of B . bronchiseptica infection . The opposite was found for the Hungarian sows and their piglets . B . bronchiseptica infection commenced at 3 and 4 weeks of age in piglets of young and old Dutch sows, respectively, followed by the emergence of P . multocida infection at 5 (YS) and 6 weeks of age (OS) . In Hungarian piglets, B . bronchiseptica infection was first demonstrable at 1 (YS) and 3 (OS) while P . multocida infection at 3 (YS) and 5 (OS) weeks of age . The serological tests demonstrated higher B . bronchiseptica specific antibody levels in the Dutch sows and piglets as compared to the Hungarian ones . According to the ELISA results, the levels of IgA and IgG in the serum and those of sIgA, IgA and IgG in the nasal secretion of Dutch sows were significantly (p < 0.001) higher in the Dutch than in the Hungarian piglets up to 3 and 4 weeks of age, respectively.

Tijdschr Diergeneeskd, 1993 Aug 1, 118(15), 469 - 71
{Pasteurella anatipestifer: a controllable farm problem}; de Wit JJ et al.; The performance of 13 flocks of ducks on a duck farm decreased markedly . Post-mortem and bacteriological examinations indicated that Pasteurella anatipestifer was a major cause, although Salmonella spp., Escherichia coli and Treponema spp . were also detected . Use of an autovaccine against Pasteurella anatipestifer markedly reduced the signs and symptoms in the second part of fattening period.

Zentralbl Bakteriol, 1993 Aug, 279(3), 387 - 93
In vitro susceptibility of Pasteurella multocida subspecies multocida strains isolated from swine to 42 antimicrobial agents; Gutierrez Martin CB et al.; The minimal inhibitory concentrations (MICs) of 42 antimicrobial agents were determined against 59 strains of Pasteurella multocida subspecies multocida, all isolated from swine lungs with lesions indicative of pneumonia . Penicillins (except cloxacillin), aminoglycosides, tetracyclines, erythromycin, josamycin, thiamphenicol, colistin, rifampin and mupirocin showed good activities, with ranging resistance between 0 and 6.8% . Higher resistance was observed for spiramycin and fosfomycin . Tylosin, vancomycin, metronidazole, dapsone and tiamulin, to which strains showed high rates of resistance, were ineffective . Cephalosporins (especially the third-generation cephalosporins) and quinolones (especially the fluorinated quinolones) were the most effective antimicrobial agents against P . multocida subsp . multocida strains and they might be of value for in vivo use.

Am J Vet Res, 1993 Aug, 54(8), 1280 - 6
Toxin production by Pasteurella multocida isolated from rabbits with atrophic rhinitis; DiGiacomo RF et al.; Naturally acquired turbinate atrophy in rabbits was associated with Pasteurella multocida infection . Several in vitro and in vivo studies were conducted to document toxin production from P multocida isolates and to determine the relation of toxin to atrophic rhinitis in rabbits . Ten isolates of P multocida serotype A:12 were obtained from adult New Zealand White rabbits with noninduced atrophic rhinitis . Specific-pathogen-free rabbits inoculated intranasally with isolates of P multocida developed rhinitis and turbinate atrophy . However, inoculation with filtrates of the same bacteria failed to induce turbinate atrophy . Cytotoxicity was observed in assays, using bovine embryonic turbinate cell cultures with extracts of P multocida, but not in agar overlay cytotoxicity assays, using bovine embryonic turbinate, bovine embryonic lung, or Vero cell cultures, or in a sandwich ELISA, using monoclonal antibodies to purified P multocida toxin . Thus, turbinate atrophy was experimentally reproduced in rabbits with isolates of P multocida, but toxin was only detected in vitro by cell culture assay of P multocida extracts.

Am J Vet Res, 1993 Aug, 54(8), 1244 - 8
Variation of abscess formation in cattle after vaccination with a modified-live Pasteurella haemolytica vaccine; Littledike ET; During the spring of the first year of a vaccine study, 57 of 238 calves (24%), in which modified-live Pasteurella haemolytica vaccine (MLV) was injected twice, developed 1 or more abscesses . Abscesses were not observed after multiple visual examinations of 437 calves given killed P haemolytica bacterin or placebo injections of similar adjuvants used in the vaccine and bacterin . Calves that developed abscesses after the second injection of MLV weighed significantly (P < 0.05) less (on the basis of body weight adjusted for weaning weight) at the second injection than did those that did not develop abscesses . Compared with calves given MLV that did not develop observable abscesses, calves developing abscesses after the second injection of MLV weighed 11.0 and 14.2 kg less, respectively, at 56 days and 112 days after injection, and they had 11.0 kg less gain at 56 days after injection . Abscess prevalence tended to be highest on certain days or at certain locations used for cattle processing, and the prevalence of abscesses increased in cattle processed later on a given day . Abscesses were not observed in 2 other groups of similarly treated calves vaccinated in the autumn or in the subsequent spring.

FEMS Microbiol Lett, 1993 Aug 1, 111(2-3), 295 - 300
Structural and serological characterisation of an O-specific polysaccharide from Serratia plymuthica; Aucken HM et al.; The surface polysaccharides of a strain of Serratia plymuthica were characterised and shown to consist of a linear, acidic galactoglucomannan as well as a major and a minor neutral galactan . Immunoblotting results demonstrated cross-reactions between this strain and others with similar galactans (S . marcescens O16 and O20, Klebsiella O1, and Pasteurella haemolytica T4 and T10).

J Comp Pathol, 1993 Jul, 109(1), 71 - 81
Nasal epithelial changes induced in piglets by acetic acid and by Bordetella bronchiseptica; Gagne S et al.; Research on atrophic rhinitis of pigs has shown that both Bordetella bronchiseptica infection and experimental treatment with acetic acid predispose the nasal mucosa to colonization with Pasteurella multocida . Gnotobiotic piglets aged 3 days were dosed intranasally with either B . bronchiseptica (n = 6) or acetic acid 1 per cent (n = 10) and killed at intervals up to the 4th day after treatment . Samples of the ventral turbinates were examined by light microscopy and scanning and transmission electron microscopy . Within 12 h acetic acid induced loss of cilia, oedema, focal cell exfoliations, mitochondrial swelling and inflammatory cell infiltration . Bordetella bronchiseptica induced only a limited oedema and loss of cilia . Colonization of cilia by the bacteria was observed 96 h after infection . We conclude that, although acetic acid and B . bronchiseptica do not induce the same modifications of the nasal respiratory epithelium, their action causes stagnation of nasal mucus, which results in a nasal environment favourable to colonization by Pasteurella multocida.

Lab Invest, 1993 Jul, 69(1), 94 - 100
Receptor-mediated binding of Pasteurella multocida dermonecrotic toxin to canine osteosarcoma and monkey kidney (vero) cells; Pettit RK et al.; BACKGROUND: Binding and internalization of Pasteurella multocida dermonecrotic toxin (PMDT) by toxin-sensitive canine osteosarcoma and monkey kidney (vero) cells was examined ultrastructurally . EXPERIMENTAL DESIGN: Purified PMDT was conjugated to 20 nm colloidal gold particles in order to observe binding and internalization in the two cell lines at the ultrastructural level . The effects of various compounds on PMDT-vero binding were investigated to help elucidate the nature of putative vero cell receptors . RESULTS: Colloidal gold-labeled PMDT was located at cell surfaces within 1 minute of its addition and rapidly transported to coated and noncoated invaginations of the plasma membrane . After extended incubation, gold particles were observed in endocytic vesicles, but not in any other intracellular structures . The magnitude of gold-PMDT cell association correlated with the cytotoxic sensitivity of the two cell lines . Early, but not late, addition of the lysosomotropic agent methylamine protected vero cells from the cytotoxic effects of PMDT without affecting binding . Biochemical and ultrastructural inhibition studies suggested the requirement for a ganglioside-type vero cell receptor . CONCLUSIONS: This is the first report describing binding and internalization of PMDT in host cells . Biochemical and ultrastructural results suggest that PMDT interacts with a ganglioside-type receptor on vero cells and is transported to the cytosol in endocytic vesicles which do not appear to fuse with lysosomes.

Res Vet Sci, 1993 Jul, 55(1), 85 - 91
Vaccine-derived Pasteurella haemolytica alter the responses of ovine pulmonary artery and vein to drugs acting on adenylate cyclase; Weekley LB et al.; Sheep were vaccinated with a live, vaccine-derived strain of Pasteurella haemolytica . The ex vivo response of isolated pulmonary artery and vein to isoproterenol, cholera toxin, sodium fluoride and calcium were examined three days after vaccination . In the pulmonary artery (endothelium intact), vaccination did not alter the response to isoproterenol, or sodium fluoride whereas the relaxation response to cholera toxin was impaired . In the pulmonary artery (endothelium removed), the maximum relaxation attained in response to isoproterenol was reduced and the response to exogenous calcium, sodium fluoride and cholera toxin not altered . In the pulmonary vein (endothelium intact), the response to isoproterenol and sodium fluoride was unchanged whereas the response to cholera toxin was impaired . In the pulmonary vein (endothelium removed), the response to isoproterenol and sodium fluoride was not altered following P haemolytica vaccination whereas the relaxation response to cholera toxin was enhanced and the response to exogenous calcium slightly impaired . These experiments suggest that vaccination with live strains of P haemolytica cause subclinical disturbances in the pulmonary circulation and may potentially alter the animals' response to pathogens.

Can J Vet Res, 1993 Jul, 57(3), 198 - 203
Effects of Pasteurella haemolytica culture supernate on bovine tracheal smooth muscle; Belanger A et al.; Pasteurella haemolytica leukotoxin is a ruminant specific leukotoxin that has been implicated in the pathogenesis of shipping fever in cattle . The present study was undertaken to determine the effect of this toxin on bovine airway smooth muscle . In vitro, the addition of culture supernate containing leukotoxin to bovine tracheal smooth muscle resulted in contraction of 55% of the muscle strips tested . Maximum responses were reached rapidly during cumulative additions of this material . In 95% of the muscle strips that responded, maximum responses were obtained after the addition of one or two cumulative doses . Repeated additions of culture supernate resulted in decreased responsiveness . Since responsiveness to other agonists was not affected, these results suggest the development of a condition similar to tachyphylaxis . The contractions were inhibited by antihistamines . Diphenhydramine, at a concentration of 10(-6) M (dose-ratio 7), and mepyramine, at a concentration of 2 x 10(-7) M (dose-ratio 56), blocked the contractions by 84% and 100% respectively . In addition, the contractions were blocked by the muscarinic antagonist atropine, but this inhibition was much weaker (46%) and was present at high concentrations only . Inhibition of the contractions by H1 receptor antagonists suggests that the contractions are mediated via H1 receptors . Since the dose-response relationship is not typical of a drug-receptor interaction, it appears unlikely that the leukotoxin is a direct agonist of H1 receptors . It is proposed that an indirect mechanism of action involving the release of histamine by tissue mast cells is responsible for the leukotoxin-induced contractions.

Can J Vet Res, 1993 Jul, 57(3), 190 - 7
Actinobacillus pleuropneumoniae culture supernatants interfere with killing of Pasteurella multocida by swine pulmonary alveolar macrophages; Chung WB et al.; The effect of Actinobacillus pleuropneumoniae culture supernatant on swine pulmonary alveolar macrophage (PAM) functions was studied . The A . pleuropneumoniae culture supernatant was toxic to PAMs when tested by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) and lactate dehydrogenase (LDH) release assays . Biological activity of the supernatant was ascribed to cytotoxins . Both the LDH and MTT assays were used for measurement of crude A . pleuropneumoniae cytotoxin concentration with good reproducibility . A preparation containing 6,800 toxic units/mL (determined by MTT assay) was used for subsequent experiments . The objective was to study the effect of crude cytotoxin on the ability of swine PAMs to kill Pasteurella multocida . Phagocytosis of opsonized P . multocida type A by PAMs was not efficient . Only 8% of incubated organisms were ingested by noncytotoxin-treated PAMs after 30 min phagocytosis . The bactericidal effect of noncytotoxin-treated PAMs only last for 60 min, after which, the rate of growth of surviving P . multocida exceeded the rate of bacterial killing by PAMs . Complete elimination of P . multocida by PAMs was not observed in this study . A total loss of ability to kill P . multocida by PAMs was seen when the PAMs were pretreated with a high concentration (340 toxic units/mL) of A . pleuropneumoniae cytotoxin . If the PAMs were pretreated with a low concentration (3.4 toxic units/mL) of cytotoxin, a significant reduction in the killing of P . multocida was still observed . The reductions in phagocytosis, phagosome-lysosome fusion (demonstrated using yeast particles of Candida albicans), and oxidative burst (demonstrated by nitro blue tetrazolium reduction (NBT) assay) may have contributed to the impaired killing of P . multocida by PAMs.(ABSTRACT TRUNCATED AT 250 WORDS)

Can J Vet Res, 1993 Jul, 57(3), 159 - 65
The (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium) colorimetric assay for the quantitation of Actinobacillus pleuropneumoniae cytotoxin; Chung WB et al.; Using swine neutrophils as target cells, two MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium) colorimetric assay systems, one with and one without phorbol 12-myristate 13-acetate (PMA) stimulation were established for the quantitation of Actinobacillus pleuropneumoniae cytotoxin . The MTT assays were optimized for the number of neutrophils, incubation time, and PMA concentration by a series of experiments . The optimal conditions were 25 x 10(4) cells/well incubated for four hours for the assay system without PMA stimulation, and 12.5 x 10(4) cells/well incubated for two hours for the assay system with PMA stimulation . One culture supernatant of a toxigenic Pasteurella multocida strain and five A . pleuropneumoniae cytotoxin preparations produced from three A . pleuropneumoniae strains were used to test assay reproducibility . Results showed both assays were reproducible with a coefficient of variation ranging from 7.8 to 18% for the assay system without PMA stimulation and from 10.7 to 18.2% for the assay system with PMA stimulation . The PMA-stimulated assay had 40 to 60-fold higher sensitivity than the nonstimulated MTT assay . The MTT assay also was applied to the measurement of neutralizing antibody titers against A . pleuropneumoniae cytotoxin.

Avian Dis, 1993 Jul-Sep, 37(3), 908 - 11
Pasteurella anatipestifer-like bacteria associated with respiratory disease in pigeons; Andreasen JR Jr et al.; Two cases of respiratory disease in pigeons are described . The first case involved pneumonia and tracheitis, and the second case involved tracheitis . In both cases, unusual gram-negative, non-fermenting, short rod-shaped bacteria were recovered along with other microorganisms . The bacteria produced small, glistening, gray colonies on blood agar, did not grow on MacConkey agar, were unreactive on several biochemical tests, and resembled Pasteurella anatipestifer . Neither pigeon isolate was distinguished from P . anatipestifer by biochemical tests . However, there were morphologic and growth differences between the pigeon isolates and P . anatipestifer . Furthermore, unlike P . anatipestifer, both pigeon isolates were sensitive to aminoglycoside antibiotics and to polymyxin B . Finally, neither isolate was agglutinated by antisera to 15 serotypes of P . anatipestifer . Diagnosticians, especially those who seldom encounter P . anatipestifer, might have difficulty distinguishing the pigeon isolates from P . anatipestifer because of their close resemblance.

Avian Dis, 1993 Jul-Sep, 37(3), 781 - 5
Pathogenicity of Pasteurella multocida: its variable nature demonstrated by in vivo passages; Matsumoto M et al.; Pasteurella multocida serotype 3,4 was isolated from a dead turkey, and the variable nature of its pathogenicity was demonstrated after in vivo passages . The original isolate was encapsulated, and its mean infectious dose (ID50) was higher than 10(8.2) colony-forming units (CFU) . To increase virulence, the organism was passaged intravenously in turkeys . After five passages, the encapsulated organism caused 67% mortality with a 10(2) CFU dose, and 50% of the contact control birds also died . A non-encapsulated variant that developed from the original isolate resulted in no mortality, even at a dose as high as 10(9) CFU . After four intratracheal passages, however, the virulence of the non-encapsulated variant increased (ID50 approximately 10(6) CFU), despite no apparent change in its morphological characteristics . These results suggest that both encapsulated and non-encapsulated forms of P . multocida can increase their pathogenicity by bird-to-bird transmission in a short period of time.

Avian Dis, 1993 Jul-Sep, 37(3), 756 - 62
Cellular defense of the avian respiratory system: dose-response relationship and duration of response in intratracheal stimulation of avian respiratory phagocytes by a Pasteurella multocida bacterin; Toth TE et al.; In experiments analyzing dose-response, intratracheal inoculation of chickens with 10(8) and 10(9) avirulent Pasteurella multocida organisms induced the migration within 24 hr of large numbers of respiratory lavage cells (RLC) with increased phagocytic proportions and phagocytic capacity . Doses from 10(4) to 10(7) organisms per bird resulted in elevated numbers of RLCs that were not significantly higher (P > 0.05) than values of uninoculated or mock-inoculated control chickens . When analyzing the duration of response, we found that inoculation with 10(9) organisms resulted in significantly higher (P < 0.05) numbers of RLCs for 63 to 89 hr without significant elevation in phagocytic proportion and capacity . The numbers of RLCs were elevated, although not significantly (P > 0.05), up to 11 days after inoculation . These results indicate that RLCs migrate to the respiratory tract only in response to a relatively high number of stimulating bacterial organisms and that the duration of response is relatively short . Although there were elevated numbers of RLCs beyond 89 hours after stimulation, the question remains as to whether these cells would enhance nonspecific defense of the respiratory system of chickens.

Avian Dis, 1993 Jul-Sep, 37(3), 668 - 72
IgA, IgG, and anti-Pasteurella multocida antibody levels in bursectomized and/or cyclophosphamide-treated turkeys after CU vaccination; Schlink GT et al.; In bursectomized, cyclophosphamide-treated, and bursectomized/cyclophosphamide-treated turkeys, IgA, IgG, and anti-Pasteurella multocida were determined before and after vaccination with the Clemson University (CU) strain of P . multocida . Before vaccination, the average total serum level of IgA was significantly (P < 0.05) lower in bursectomized and bursectomized/cyclophosphamide-treated turkeys than in untreated controls, and the average serum levels of anti-P . multocida were significantly (P < 0.05) lower in bursectomized, cyclophosphamide-treated, and bursectomized/cyclophosphamide-treated turkeys than in untreated controls . After vaccination, average serum IgA levels were still significantly (P < 0.05) lower in all treated groups of turkeys than in the untreated controls . Also after vaccination, total IgG increased significantly (P < 0.05) only in the bursectomized turkeys, and serum anti-P . multocida antibody levels increased significantly (P < 0.05) in cyclophosphamide-treated, bursectomized/cyclophosphamide-treated, and untreated turkeys . After challenge with virulent P . multocida, survivability was significantly (P < 0.05) lower in the three treated groups of turkeys than in the untreated groups.

J Biol Chem, 1993 Jun 15, 268(17), 12764 - 74
Characterization of a specific ligand for P-selectin on myeloid cells . A minor glycoprotein with sialylated O-linked oligosaccharides; Norgard KE et al.; Lectin-carbohydrate recognition between the selectins and their ligands are among the earliest events in leukocyte recirculation, leukocyte recruitment into inflamed areas, and abnormal egress of leukocytes in diseases . Previously, we have described a dimeric sialoglycoprotein from myeloid cells with subunits of molecular mass = 120 kDa, which is selectively recognized by P-selectin (Moore, K.L., Stults, N.L., Diaz, S., Smith, D.F., Cummings, R.D., Varki, A., and McEver, R.P . (1992) J . Cell Biol . 188, 445-456) . Here, we demonstrate that this P-selectin ligand carries alpha 2-3-linked sialic acids and the sialyl-Lewisx (SLex) tetrasaccharide motif . This glycoprotein contains < 1% of the total membrane-bound sialic acids and a very small fraction of the total SLex on neutrophil membranes . In spite of a relative resistance to sialidase digestion, the predominant form of sialic acid on the ligand is N-acetylneuraminic acid . Selective periodate oxidation of the side chain of sialic acids does not affect P-selectin binding and allows the introduction of tritium label into the truncated sialic acids . beta-Elimination with alkaline borohydride releases labeled O-linked oligosaccharides both from the labeled neutrophil ligand and from the ligand purified from HL-60 cells metabolically labeled with {3H}glucosamine . The ligand from both neutrophils and HL-60 cells is also susceptible to cleavage by the enzyme O-sialoglycoprotease from Pasteurella hemolytica . Analysis of the specificity of this enzyme suggests that the P-selectin ligand carries large numbers of closely spaced sialylated O-linked oligosaccharides . O-Sialoglycoprotease abolishes both direct binding of P-selectin to HL-60 cells and the adhesion of HL-60 cells to immobilized P-selectin, without significantly decreasing overall cell surface SLex expression . This indicates that the 120-kDa ligand may be the major determinant of P-selectin:myeloid cell interaction in vivo . Finally, based on the current and previous data, we hypothesize that the high affinity recognition site(s) of this P-selectin ligand may be derived from a "clustered saccharide patch" of sialylated fucosylated O-linked oligosaccharide sequences.

J Infect Dis, 1993 Jun, 167(6), 1281 - 7
Effects of morphine dependence on the pathogenesis of swine herpesvirus infection; Risdahl JM et al.; To further understand the effects of opiates on the pathogenesis of infectious disease, naturally occurring pathogens were studied in a swine model . Swine were given morphine for 21-42 days to establish a tolerant, dependent state . On day 7 after morphine initiation, pigs were challenged with swine herpesvirus-1 (SHV-1); on day 14, selected animals were superinfected with Pasteurella multocida . Evaluations were made of the clinical disease, protective effect of SHV-1 vaccination, and pathology . Morphine-dependent animals developed significantly greater virus-induced and secondary bacterial pneumonia . Prior vaccination with SHV-1 was not protective against pneumonia in morphine-dependent pigs . Unexpectedly, clinical signs associated with neurologic disease were less pronounced, and mortality from viral encephalitis was decreased in morphine-treated animals . Collectively, the findings demonstrate that morphine dependence is associated with a marked alteration of the pathogenesis of SHV-1 and that the effects of this opiate on pathogenesis are determined by the specific site of infection.

Infect Immun, 1993 Jun, 61(6), 2618 - 25
Enhancement of neutrophil-mediated injury to bovine pulmonary endothelial cells by Pasteurella haemolytica leukotoxin; Maheswaran SK et al.; In this study, we used an in vitro coculture system to determine which virulence factor from Pasteurella haemolytica A1 was responsible for augmenting bovine polymorphonuclear neutrophil (PMN)-mediated killing of bovine pulmonary artery endothelial cells (BPAEC) . A 51Cr release cytotoxicity assay was used as a measure of BPAEC killing . The mechanisms associated with this BPAEC killing were also studied . Our results demonstrated that the leukotoxin and not the lipopolysaccharide from P . haemolytica was responsible for augmenting the PMN-mediated killing of BPAEC . Furthermore, this augmented killing was related to the stimulation of PMNs by the leukotoxin . Killing of BPAEC by leukotoxin-stimulated PMNs was diminished in the presence of the H2O2 inactivator, catalase . The membrane-permeant H2O2, hydroxyl radical (HO.) scavenger 1,3-dimethyl-2 thiourea, and the HO . scavenger dimethyl sulfoxide but not the myeloperoxidase inhibitor sodium azide attenuated this BPAEC killing . Pretreatment of BPAEC with a 21-aminosteroid (U74500A), a potent iron chelator-antioxidant, provided the most effective protection against BPAEC killing induced by leukotoxin-stimulated PMNs . These data were compatible with the concept that the H2O2 generated by leukotoxin-stimulated PMNs interacts with intracellular iron in the endothelial cell to form highly reactive HO. . We suggest that HO . may be a key factor in BPAEC killing . Furthermore, since the elastase-specific inhibitor N-methoxy-succinyl-Ala-Ala-Pro-Val-chloromethyl ketone (CMK) also attenuated BPAEC killing and both CMK and 1,3-dimethyl-2 thiourea functioned additively in protecting against BPAEC killing, we conclude that both HO . and elastase may jointly contribute to BPAEC killing induced by leukotoxin-stimulated PMNs . This study broadens our understanding of how leukotoxin-stimulated PMNs injure lung endothelial cells and provides new insight into the pathogenesis of bovine pneumonic pasteurellosis.

Calcif Tissue Int, 1993 Jun, 52(6), 455 - 9
Purified Pasteurella multocida protein toxin reduces acid phosphatase-positive osteoclasts in the ventral nasal concha of gnotobiotic pigs; Ackermann MR et al.; To study the in vivo response of conchal (turbinate) osteoclasts to Pasteurella multocida toxin, four gnotobiotic pigs (7 days of age) were inoculated subcutaneously with 0.2 microgram/kg of purified toxin . One toxin-treated pig along with one control pig were necropsied at 2, 5, 9, and 14 days postinoculation . The entire length of nasal concha from the nasal planum toi ethmoid region was removed, blocked by transverse cuts into five areas, decalcified, sectioned, and then stained with tartrate-resistant acid phosphatase (TRAP) to identify osteoclasts . In each section, total area of concha, total osteoclast cytoplasmic area, and number of osteoclasts were determined using an image analysis morphometric unit . Also collected from pigs were blood and serum for complete blood counts, electrolyte levels, liver enzymes, and TRAP levels . Conchal atrophy increased in severity with time after 2 days postinoculation . In general, the ventral conchae from toxin-treated pigs at 9 and 14 days postinoculation had decreased surface area, osteoclast cytoplasmic area, and numbers of osteoclasts . Serum levels of TRAP were mildly elevated when compared with age-matched controls . No other significant alterations in blood cells or chemistries occurred and no lesions were present histologically in tissues (liver, kidney, lung, heart, and spleen) other than concha . This study shows that the P . multocida toxin induces rapid bone resorption and increases serum levels of acid phosphatase but leads to diminished acid phosphatase expression and presumably, numbers of osteoclasts.

Zentralbl Bakteriol, 1993 Jun, 279(1), 7 - 26
Ecology and significance of Pasteurellaceae in animals; Bisgaard M; The reservoir of eighty-one taxa/groups classified with the family Pasteurellaceae Pohl 1981 is reviewed based upon published data and own investigations . With the exception of certain strains of P . multocida, A . pleuropneumoniae and {H.} paragallinarum organisms belonging to this family are usually regarded as opportunistic, secondary invaders which under normal conditions coexist peacefully with the animal host on mucosal membranes of the upper respiratory- and lower genital tracts . Very little is known about factors that govern the ecological preferences that certain members of this family show for specific surfaces and hosts . Mechanisms of colonization, survival and multiplication, invasion and pathogenic action are incompletely understood . The significance of Pasteurellaceae in animals and man has recently been reviewed . Subsequent publications have underlined the significance of biovars 2 of P . canis and P . avium and ornithine negative P . multocida in pneumonia in cattle . In addition, differences in pathogenicity have been demonstrated for different serovars of {H.} parasuis . The disease potential of many taxa/groups is only incompletely known.

Zentralbl Bakteriol, 1993 Jun, 279(1), 45 - 50
Detection of an adenylate cyclase gene in Pasteurella species; Escande F et al.; A Pasteurella multocida adenylate cyclase gene has been previously cloned in Escherichia coli and sequenced . A 1200 bp HpaI fragment from the coding region was used as a probe to analyse the presence of the gene in different Pasteurella species and subspecies, Actinobacillus ureae (formerly P . ureae) and group EF-4 bacteria . Thirty-seven strains were checked for the presence of the gene . It was shown that the adenylate cyclase gene was detected only in the species Pasteurella multocida.

Zentralbl Bakteriol, 1993 Jun, 279(1), 131 - 9
Epidemiology of human infections by Pasteurella and related groups in France; Escande F et al.; A retrospective study of infections due to Pasteurella (P.) and related groups was performed between the Pasteurella National Center and Nancy's hospital from 1985 to 1991 . Among the 958 cases recorded, wound infections (bites, scratches and punctures) were the common forms of pasteurellosis (66%) caused by P . multocida (48%), P . canis (11%), P . dagmatis (5%), P . stomatis (4%), and in few cases by groups EF-4 and M-5 (14 and 13%, respectively) . In human infections unrelated to animal wounds, respiratory tract diseases and bacteremia-septicemia were the predominant infections with respectively 19 and 11%, and caused by P . multocida . Next in importance were urogenital (2.5%), abdominal (1%) and central nervous system (< 1%) infections . The majority of animal bite wound infections was treated with penicillins or tetracyclines; with other forms, penicillins and cephalosporins were more likely.

Zentralbl Bakteriol, 1993 Jun, 279(1), 125 - 30
Classification of Pasteurella field strains isolated from farms in Germany using traditional methods and DNA-DNA hybridization; Schimmel D et al.; 410 Pasteurella (P.) field strains isolated from calves and piglets were classified according to Bisgaard et al . (1) . 376 strains were assigned to P . multocida ssp . multocida, 34 of them were ornithine- and trehalose+, and 61 of them ornithine- and trehalose- . 4 strains belonged to P . multocida ssp . septica, 4 to P . multocida ssp . gallicida, 6 to P . avium biovar 2 and 20 to P . canis biovar 2 . There was no difference in the prevalence of the species in calves and pigs . The fact that strains belonging to P . multocida ssp . septica were isolated only from calves and P . multocida ssp . multocida ornithine- and trehalose- were mostly isolated from piglets could indicate a certain host specificity of these isolates . In genotypic investigations 20 field isolates of P . multocida belonging to different Carter serotypes, as well as serologically negative strains were compared to reference strains in terms of deoxyribonucleic acid (DNA) relatedness . The data obtained by filter hybridization revealed a considerable degree of genotypic intraspecies heterogeneity within P . multocida . No correlations to the respective serotypic classification could be detected.

J Vet Med Sci, 1993 Jun, 55(3), 455 - 6
An occurrence of equine transport pneumonia caused by mixed infection with Pasteurella caballi, Streptococcus suis and Streptococcus zooepidemicus; Hayakawa Y et al.; An acute death occurred in a racehorse with pneumonia after long-distance transportation in December, 1990 . Pasteurella caballi, Streptococcus suis and Streptococcus zooepidemicus were isolated from the lung at high rate . Specific antigens of these bacteria were also demonstrated immunohistologically in the pneumonic lesion . These findings indicated that the disease is equine transport pneumonia caused by a mixed infection of the three bacterial species . This is the first report on the isolation of P . caballi and S . suis from a racehorse in Japan.

Lab Anim Sci, 1993 Jun, 43(3), 217 - 21
Pathologic features associated with decreased longevity of mutant sphha/sphha mice with chronic hemolytic anemia: similarities to sequelae of sickle cell anemia in humans; Grossmann A et al.; A colony of sphha/sphha mice with congenital hemolytic anemia and an abnormality in erythrocyte spectrin assembly was screened to determine the cause of premature death . Sphha/sphha mice have decreased life span, with 50% of animals dying by 6 months of age . The phenotype of these mutant mice includes moderate anemia (hematocrit: 21 to 28%), reticulocytosis, leukocytosis, lymphocytosis, extensive extramedullary hematopoiesis in spleen and liver, lymph node hyperplasia and membranoproliferative glomerulonephritis . With increased surveillance of this mouse colony, 20 clinically sick anemic mice were evaluated (complete blood counts and cultures of blood), euthanized and necropsied . Compared with anemic mice without clinical signs of disease, sick anemic mice had significantly higher white blood cell counts with only 4 (20%) of 20 animals being severely anemic (hematocrit: 4 to 8%) . Blood from 11 (45%) of 20 animals was culture-positive for Pasteurella pneumotropica, Enterococcus, and/or Escherichia coli . In addition to the usual lesions in sphha/sphha mice, sick anemic mice had pneumonitis (95%) with thrombosis and infarction (80%) of one or more organs (spleen, myocardium, pancreas, liver, or bone marrow) . The thrombotic tendency that accompanies the chronic hemolytic anemia in sphha/sphha mice, as well as the other clinicopathologic changes in these mutant mice, bears a striking resemblance to some poorly understood sequelae in human patients with sickle cell anemia . This mouse model may be useful in studying the pathophysiology of complications associated with sickle cell anemia in humans.

Berl Munch Tierarztl Wochenschr, 1993 Jun, 106(6), 194 - 7
{Detection of toxin-producing types of Pasteurella multocida--a comparison of methods}; Erler W et al.; 28 Pasteurella multocida strains were examined for production of toxin by 6 different methods . Identical results were obtained using a mice lethality test, a tissue cell culture assay and an ELISA . Different results were received with a dot-blot-immunoassay (1 strain), using a gene probe (6 strains) and a guinea pig skin test (8 strains) . Corresponding differences were detected with 2 strains only . Tissue culture, ELISA and dot-blot-immunoassay are effective methods for the diagnosis of toxin-producing Pasteurella multocida strains . Animal experiments should be an exception.

Am J Vet Res, 1993 Jun, 54(6), 897 - 900
Prevalence of mycoplasmal and ureaplasmal recovery from tracheobronchial lavages and of mycoplasmal recovery from pharyngeal swab specimens in cats with or without pulmonary disease; Randolph JF et al.; The prevalence of mycoplasmal and ureaplasmal recovery from tracheobronchial lavage specimens and prevalence of mycoplasmal recovery from pharyngeal swab specimens from cats with (28) or without (18) pulmonary disease were determined . Mycoplasmas were recovered from tracheobronchial lavage specimens in 21% of cats with pulmonary disease, but in no cats without pulmonary disease; this difference is significant (P = 0.04) . Mycoplasmal recovery from tracheobronchial lavage specimens was not significantly associated with concurrent Pasteurella spp isolation, septic inflammation, or bronchitis . Ureaplasmas were only isolated from a tracheobronchial lavage specimen in 1 cat with pulmonary disease and in no cats without pulmonary disease . Similar mycoplasmal recovery rates were found for pharyngeal swab specimens from cats with (39%) or without (35%) pulmonary disease . Seemingly, mycoplasmas are part of the normal pharyngeal flora in approximately a third of the feline population, but mycoplasmas are not normal inhabitants of the lower respiratory tract in cats . It is unknown whether mycoplasmas isolated from tracheobronchial lavage specimens in cats with pulmonary disease are primary pathogens or opportunistic invaders . Seemingly, ureaplasmas are seldom associated with pulmonary disease in cats, and are not normal inhabitants of the trachea and bronchi of cats.

Am J Vet Res, 1993 Jun, 54(6), 891 - 6
Effects of ampicillin and trimethoprim-sulfamethoxazole on the vaginal bacterial flora of bitches; Strom B et al.; Vaginal aerobic bacterial flora was studied in 5 healthy bitches before, during, and after a 10-day period of treatment with ampicillin and an equally long period of treatment with trimethoprim-sulfamethoxazole . Blood variables and antimicrobial drug susceptibility also were studied . Bacteria were isolated from all bitches before the first treatment period . Bitches from which only a sparse number of bacteria were isolated had flora that varied from day to day . In most instances when bitches were given an antibiotic to which their vaginal bacterial flora was susceptible, these bacteria were eradicated after only 1 day of treatment . This was true for pasteurella, streptococci, and, in all but one case, Escherichia coli . Staphylococcus intermedius was more difficult to eradicate, and, although susceptible in vitro, it was unaffected by antibiotic treatment in 1 bitch and it took 7 days to eradicate in another . Eradication of aerobic bacteria in the vagina was total only in the bitch that had sparse flora from the beginning . Bacteria colonized within 0 (in 4/5 bitches) to 4 days after termination of treatment with ampicillin and within 0 (in 4/5 bitches) to 3 days for trimethoprim-sulfamethoxazole . Mycoplasmas emerged during and after both treatment periods, and E coli became apparent during treatment with trimethoprim-sulfamethoxazole . Because mycoplasmas may be genital pathogens in bitches and E coli is a common uropathogen, their appearance should be an argument against widespread use of antibiotics in healthy breeding bitches . Two bitches developed a vaginal discharge during treatment or shortly after . Blood variables did not change during the study, nor did antimicrobial drug resistance of the isolated bacteria.

Am J Vet Res, 1993 Jun, 54(6), 856 - 61
Serum antibody response to purified Pasteurella haemolytica capsular polysaccharide in cattle; Tigges MG et al.; Capsular polysaccharide (CP) of Pasteurella haemolytica, biotype A, serotype 1, was purified and combined with saline solution, aluminum hydroxide, and Freund's incomplete (oil) adjuvant . Three groups of calves were administered the various antigen preparations . The CP in saline preparation was also administered to 5 mature cows . Second injections were given 4 weeks after the first . Weekly obtained serum samples were analyzed for P haemolytica-specific antibody, using the indirect hemagglutination assay, and CP-specific antibodies were detected, using an isotype-specific ELISA . Purified CP stimulated production of CP-specific IgM, IgG1, and IgG2 in calves and predominantly IgM and IgG1 in mature cows . Significant increases in CP antibody titers were not observed after the second injection of CP antigen in either calves or mature cows . The CP in oil adjuvant stimulated the highest mean CP-specific IgG1 and IgG2 responses, whereas the CP in aluminum hydroxide adjuvant stimulated the highest mean CP-specific IgM response.

Singapore Med J, 1993 Jun, 34(3), 271 - 3
Pasteurella multocida septicaemia following a dog bite; Sin Fai Lam KN et al.; Bite wounds are often mistakenly considered innocuous . However, they are frequently complicated by infection which may be serious . We describe a case of Pasteurella multocida septicaemia with myopericarditis following a dog bite . Treatment of the infection as well as active support of myocardial function led to a successful outcome.

Zentralbl Bakteriol, 1993 Jun, 279(2), 259 - 73
Hemagglutination by Pasteurellaceae isolated from rodents; Boot R et al.; Pasteurellaceae notably P . pneumotropica, have been associated with severe outbreaks of respiratory disease in several species of rodents . Host-specific parasitism of Pasteurellaceae in rodents has hardly been studied . Since host tropism in many bacteria involves adhesive mechanisms, we examined the hemagglutinating (HA) properties of 44 isolates from different rodent species (mouse (15) rat (8), hamster (9), gerbil (10) and Mastomys (2)) . Only 13 mouse isolates and the 2 Mastomys isolates hemagglutinated human (type O Rh+) and canine red blood cells (RBCs) . No HA was found using RBCs from 10 other animal species . HA was not inhibited by simple sugars and glycoconjugates, but was completely inhibited by heating of bacterial cells for 10 min at 80 or 100 degrees C, partially inhibited by glutaraldehyde and inhibited in a dose-dependent mode by NaIO4, suggesting the involvement of bacterial polysaccharide structures in the HA process . Enrichment procedures did not reveal the presence of HA- subpopulations in HA+ isolates or the presence of HA+ subpopulations in HA- isolates . Electron microscopy revealed the presence of fimbriae both in HA+ and HA- isolates . A regularly structured (RS) layer was detected on cells of part of the HA+ isolates only . Our results suggest that Pasteurellaceae of mice and Mastomys may be related and differ from isolates isolated from other rodent species.

Zentralbl Bakteriol, 1993 Jun, 279(1), 75 - 82
Phenotypical characters and ribotyping of Pasteurella aerogenes from different sources; Lester A et al.; On the occasion of five Danish human Pasteurella aerogenes from pig bite lesions, a comparison was made between 6 isolates from man and 15 animal isolates, mainly from pigs . The strains originated from 6 different countries (USA, Canada, Czechoslovakia, France, Belgium and Denmark) . The 21 isolates were characterized by conventional biochemical tests, antibiogram and the API 20 NE kit; finally ribotyping was carried out by hybridizing EcoRI-digested chromosomal DNA with a probe derived from E . coli ribosomal RNA . By ribotyping, 19 of the 21 strains clustered at a similarity level of 81% or more; both phenotypical tests and ribotyping indicated that the remaining two strains did not belong to the species P . aerogenes . In conclusion, despite minor differences our P . aerogenes isolates constituted a well-defined group and they could not be subdivided on basis of animal or geographical origin.

Ophthalmic Surg, 1993 May, 24(5), 346 - 8
Pasteurella multocida keratitis and corneal laceration from a cat scratch; Ho AC et al.; A 24-year-old woman was evaluated 12 hours after she sustained a cat scratch to her left eye . Slit-lamp examination revealed a Seidel-positive corneal laceration with a surrounding dense full-thickness corneal ulcer and severe inflammatory reaction . Since the anterior chamber was well formed, it was decided not to repair the laceration on an emergency basis . She was initially treated with intensive topical fortified tobramycin and vancomycin, and intravenous gentamicin and clindamycin . Cultures of the corneal ulcer revealed Pasteurella multocida and the antibiotic regimen was adjusted appropriately . The laceration healed without surgery, and the infection resolved well, with excellent visual acuity.

DNA Cell Biol, 1993 May, 12(4), 351 - 62
Molecular analysis of the Actinobacillus pleuropneumoniae RTX toxin-III gene cluster; Chang YF et al.; Actinobacillus pleuropneumonia strains that secrete three different exotoxins (ApxI, ApxII, and ApxIII) have been implicated in the etiology of porcine pleuropneumonia . To understand the role of these toxins in the pathogenesis of this disease, we have previously reported the cloning of the hemolysin gene (apxII) (Chang et al., 1989a), which encodes a 110-kD polypeptide with hemolytic and cytotoxic activity . To clone the third toxin gene (apxIII), a new genomic library using A . pleuropneumoniae serotype 2 chromosomal DNA was constructed . A series of five overlapping recombinant phage clones carrying the gene (apxIII) for this 120-kD antigen were identified using a DNA probe containing sequences from the Pasteurella haemolytica lktBD genes . Sequence analysis of a region of the cloned DNA reveals four open reading frames encoding proteins with predicted masses of 20.4, 112.5, 80.3, and 54.7 kD . These genes, designated apxIIC, apxIIIA, apxIIIB, and apxIIID, respectively, are similar in sequence to the RTX (repeat of toxin) toxin family . The toxin produced by the cloned gene kills BL-3 cells and is not hemolytic in vitro.

Proc Natl Acad Sci U S A, 1993 May 1, 90(9), 4211 - 5
Functional replacement of the hemolysin A transport signal by a different primary sequence; Zhang F et al.; Secretion of the 107-kDa hemolysin A (HlyA) from Escherichia coli is mediated by the membrane proteins hemolysin B and hemolysin D . Hemolysin B is a member of the so-called ATP binding cassette transporter superfamily, which includes the multidrug resistance P-glycoprotein, the cystic fibrosis CFTR protein, and the major histocompatibility complex-associated transporter of antigenic peptides . Recognition of HlyA by the hemolysin B/D transporter is dependent on a signal sequence mapped to the C-terminal 50 or so amino acids of the HlyA molecule . We show that the C-terminal 70 amino acids of leukotoxin from Pasteurella hemolytica can substitute functionally for the HlyA signal sequence . This 70-amino acid sequence contains no primary sequence similarity to the HlyA signal sequence; however, structural motifs of helix-turn-helix followed by strand-loop-strand can be deduced for both sequences . We also demonstrate by site-directed mutagenesis that changes to these predicted motifs affect transport function . It thus appears that the transport signal of HlyA may be defined by a higher-order structure and that the hemolysin transporter may recognize a much wider diversity of primary sequences than previously anticipated . This finding may have implications for understanding the basis of substrate specificity of other ATP binding cassette transporters.

Infect Immun, 1993 May, 61(5), 2089 - 95
Molecular characterization of a leukotoxin gene from a Pasteurella haemolytica-like organism, encoding a new member of the RTX toxin family; Chang YF et al.; A Pasteurella haemolytica-like organism, a new species of bacterium isolated from piglets with diarrhea, secretes a leukotoxin into the culture media . Western blot (immunoblot) analysis indicated that this leukotoxin cross-reacted with antileukotoxin antibody derived from cattle immunized with P . haemolytica . Five overlapping recombinant bacteriophages carrying the gene for this 105-kDa polypeptide were identified with a DNA probe containing sequences from the P . haemolytica lktCA genes from a P . haemolytica-like organism strain 5943 genomic library . Sequence analysis of a region of the cloned DNA revealed two open reading frames encoding proteins with predicted masses of 19.4 and 101.6 kDa . These genes, which we designate pllktC (P . haemolytica-like organism leukotoxin C gene) and pllktA (A gene), respectively, are similar in sequence to the RTX (repeat of toxin) toxin family . The structure of the 101.6-kDa protein derived from the DNA sequence shows three transmembrane domains in the N-terminal part of the protein, 13 glycine-rich repeat domains in the second half of the protein, and a hydrophobic C-terminal part . pllktC and pllktA are strongly homologous to P . haemolytica lktC and lktA genes . However, this leukotoxin kills both BL-3 and pig leukocytes and is not hemolytic.

Antimicrob Agents Chemother, 1993 May, 37(5), 1150 - 3
Comparative susceptibilities of 173 aerobic and anaerobic bite wound isolates to sparfloxacin, temafloxacin, clarithromycin, and older agents; Goldstein EJ et al.; The in vitro activities of sparfloxacin, temafloxacin, ciprofloxacin, ofloxacin, clarithromycin, erythromycin, tetracycline, cephalothin, penicillin G, and amoxicillin-clavulanic acid against 173 recent clinical bite wound isolates were determined by agar dilution . Sparfloxacin was active against all strains (MIC for 90% of strains tested, < or = 1 micrograms/ml) except for most fusobacteria and one-third of the Prevotella spp . The other fluoroquinolones had similar activities but higher MICs, especially for streptococci . Clarithromycin was more active against many isolates including Pasteurella multocida than erythromycin, with MICs of < or = 2 micrograms/ml (versus 4 micrograms/ml for erythromycin).

J Anim Sci, 1993 May, 71(5), 1247 - 55
Effect of copper deficiency on tissue, blood characteristics, and immune function of calves challenged with infectious bovine rhinotracheitis virus and Pasteurella hemolytica; Stabel JR et al.; Fourteen Holstein steers, averaging 30 d of age, were fed a semipurified diet (1.5 mg of Cu/kg) supplemented with 0 (-Cu) or 10 mg of Cu/kg of diet (+Cu) for 5 mo . Calves were then challenged by consecutive exposure to aerosol preparations of infectious bovine rhinotracheitis virus (IBRV) and Pasteurella hemolytica on d 0 and 7, respectively, of the 30-d study . Serum ceruloplasmin and plasma copper were higher in +Cu calves throughout the challenge period and increased in +Cu calves after microbial challenge . Heart weights were higher in -Cu calves, although weights of liver, spleen, and thymus were not different between treatments . Copper concentrations in all tissues as well as thymus zinc were higher in +Cu calves . Serum immunoglobulin M tended to be higher in +Cu calves and increased in both treatments after IBRV challenge . Serum IBRV antibody titers were higher in -Cu calves with detectable seroconversion by d 10 postinfection . In contrast, antigen-specific antibodies to P . hemolytica tended to be higher in +Cu calves on d 21 . Copper status did not affect blastogenic response, but phytohemagglutinin (PHA)-stimulated blastogenesis was higher in both treatments after IBRV challenge . Repletion of lymphocyte cultures with copper chloride increased proliferative responses to PHA in both +Cu and -Cu calves, and greater responses at all levels of copper (1 to 16 micrograms/mL) were noted in -Cu calves . These results indicate that copper deficiency affects various physiological characteristics that may be important in immunological defense to pathogenic challenge.

Res Vet Sci, 1993 May, 54(3), 366 - 71
Distribution of intramuscularly administered erythromycin into subcutaneous tissue chambers before and after inoculation with Pasteurella haemolytica; Clarke CR et al.; Distribution of erythromycin into subcutaneous tissue chambers was characterised pharmacokinetically and the effect of Pasteurella haemolytica infection on the extent of penetration was studied . Thermoplastic tissue chambers were implanted subcutaneously in the paralumbar fossae of six calves . Thirty-five days after implantation, the tissue chamber distribution of intramuscularly administered erythromycin (30 mg kg-1) was studied . Chambers were then inoculated with P haemolytica and the tissue chamber pharmacokinetics of erythromycin were again studied . Diffusion of erythromycin into tissue chambers was best described using a two-compartment model with tissue chambers representing a relatively inaccessible compartment . Despite changes in chamber fluid pH, the extent of erythromycin penetration into chambers was not affected by P haemolytica inoculation . Comparison of computer simulated concentration-time curves resulting from different routes of administration revealed that penetration of erythromycin into less accessible sites was more likely to be higher after intravenous administration than after intramuscular administration.

Am J Vet Res, 1993 May, 54(5), 738 - 42
Histomorphologic features of the nasal cavity of pigs exposed to Pasteurella multocida type-D dermonecrotic toxin; Ghoshal NG et al.; Microscopic examination of the nasal mucosa of clinically normal specific-pathogen-free pigs and of toxicogenic type-D Pasteurella multocida toxin challenge-exposed specific-pathogen-free pigs indicated that the surface epithelium in pigs of both groups was microscopically normal; erosions or appreciable inflammatory changes were not evident . In pigs of both groups and in all 3 regions of the nasal cavity, the endothelial lining of all blood vessels appeared normal without detectable changes to the walls at postinoculation day 10 . Vascular injury in the cartilage or the bone was not discernible in control or challenge-exposed pigs . There were marked differences in the osseous structures of the conchae when the 2 groups were compared . In control pigs, active bone formation and remodeling were observed, and the septal cartilage was normal . In toxin challenge-exposed pigs, there likewise was normal bone formation and remodeling in the vestibular region, and the septal cartilage was normal . In marked contrast, conspicuous changes were observed in the osseous core of the conchae of the respiratory and, sometimes, the olfactory regions . These changes consisted of bone necrosis and resorption by large numbers of osteoclasts with variable replacement by dense mesenchymal stroma, which resulted in conchal atrophy . In the absence of any discernible damage or injury (angiopathy) to the nasal vessels, it appears that the action of the dermonecrotoxin of P multocida serotype D is on the most active osteoblasts and the associated organic matrix of the bone, with subsequent disruption of normal bone formation and remodeling of the nasal conchae.

Rev Med Interne, 1993 May, 14(5), 313 - 6
{Infectious diseases transmitted by animal bites}; Bricaire F; The animal population, and most of all pets, entail a high risk of bites, some of them severe, which may lead to complications among which infection is a major one . Epidemiological data about the germs liable to grow (Pasteurellae, pyogenic germs, anareobes...) are helpful to guide curative, and even more, prophylactic approaches to treatment.

Zh Mikrobiol Epidemiol Immunobiol, 1993 May-Jun, (3), 63 - 70
{The development of a technology for the production of live dried vaccines against avian pasteurellosis and swine erysipelas}; Iartsev MIa et al.; The technology of the production of dried live vaccine against Pasteurella infection of fowl from Pasteur's 2nd avirulent strain, strains AB and K, has been developed . This technology includes the process of batch cultivation of Pasteurella cells, controlled in such parameters as eH, pO2 and glucose concentration, in fermenters in optimized culture medium, based on Hottinger hydrolysate and fermentative casein-yeast hydrolysate, and preservation in improved saccharose-gelatin medium prepared in potassium sulfate buffer solution . The new technology makes it possible to increase the yield of preparations with stable biological activity 5- to 13-fold in comparison with the traditional technology . Furthermore, the technology of the production of live dried vaccine against swine erysipelas from Erysipelothrix insidiosa strain BP-2 has been developed . This technology is based on maintaining the optimum conditions of the batch cultivation of E . insidiosa in meat medium based on Hottinger hydrolysate and media obtained from hydrolysate of pancreatic fermentation products of microbial biomass; the preparation thus obtained is stabilized in peptone-saccharose-gelatin medium prepared in potassium phosphate buffer solution . This increases the yield of the vaccine 8-fold in comparison with the traditional technology, while ensuring the stability of bacteria after drying and during prolonged storage.

Am J Vet Res, 1993 May, 54(5), 695 - 700
Purification of a Pasteurella haemolytica serotype 1-specific polysaccharide epitope by use of monoclonal antibody immunoaffinity; Austin FW et al.; A murine IgM monoclonal antibody causing bacterial agglutination was used in an immunoaffinity procedure to purify a serotype 1-specific polysaccharide epitope from Pasteurella haemolytica . The P haemolytica serotype 1-specific antibody was precipitated from peritoneal ascitic fluid, dialyzed, and covalently attached to cyanogen bromide-activated Sepharose 4B beads . Retention of purified antibody activity and coupling efficiency were > 99% when evaluated by ELISA, agglutination testing, and protein determination . Potassium thiocyanate was selected as an eluant on the basis of reversible dissociation of bacterial agglutination and was titrated for the lowest effective concentration . Immunobead activity was observed microscopically by immobilization of encapsulated P haemolytica serotype 1 and its reversible dissociation after elution with 0.4M potassium thiocyanate . Specificity of immobilization was visualized, using P haemolytica serotypes 2 and 5, which were not bound, and by blocking serotype-1 binding with homologous capsular material . Saline-extractable capsular material from P haemolytica serotype 1 was used as an antigen source . After elution of the serotype 1-specific polysaccharide epitope, the product was dialyzed and analyzed, using chemical and immunologic methods . The immunoaffinity product contained no detectable protein and greater than half the original hexosamine content . Using defined monoclonal antibodies in ELISA, titration of the original capsular material and the immunoaffinity product revealed specific retention of lipopolysaccharide, a 10- to 30-kd polysaccharide antigen common to all P haemolytica and P multocida serotypes, and serotype 1-specific capsular polysaccharide, indicating possible epitope sharing among polysaccharide antigens of P haemolytica serotype 1.

Can J Vet Res, 1993 Apr, 57(2), 136 - 8
Epidemiology of Pasteurella multocida in a farrow-to-finish swine herd; Zhao G et al.; Thirty-eight clinical isolates of Pasteurella multocida, recovered from a continuous flow, farrow-to-finish swine herd, were characterized by capsular serotyping and restriction endonuclease analysis (REA) in order to study the epidemiology of P . multocida pneumonia . Twenty-three of the 38 isolates obtained in the study belonged to serotype A . They displayed three REA patterns after digestion with HpaII, of which one designated A-3 represented 70% of the samples . The remaining 15 isolates were serotype D . Four different REA patterns were observed in the type D isolates . The REA type D-1 was most prevalent and accounted for 47% of the serotype D isolates . All serotype A isolates were nontoxigenic, whereas five (33%) of the serotype D isolates were toxigenic . Vertical transmission of P . multocida could not be demonstrated, and was probably not a major route of infection . The results of this study suggest that strains of P . multocida virulent for pigs exist and cause swine pneumonic pasteurellosis in continuous flow herds by horizontal transmission.

J Am Vet Med Assoc, 1993 Apr 1, 202(7), 1106 - 10
Comparison of computed tomography with radiography as a noninvasive diagnostic technique for chronic nasal disease in dogs; Codner EC et al.; Computed tomography was evaluated as a noninvasive technique for the diagnosis of chronic nasal disease in dogs . Computed tomographic images, radiographs, and histopathologic findings were compared in 11 dogs with chronic nasal disease . Definitive diagnosis was made following traumatic nasal flush, exploratory surgery, or necropsy . The study included 8 dogs with intranasal tumors, 2 dogs with bacterial rhinitis (Pasteurella sp), and 1 dog with mycotic rhinitis (Aspergillus sp) . Computed tomography was superior to radiography in defining the extent of the disease process and in differentiating infectious rhinitis from nasal neoplasms . It defined lesions in the palate, nasopharyngeal meatus, maxillary sinus, caudal ethmoturbinates, and periorbital tissues that were difficult to demonstrate by use of conventional radiography . Tumors appeared as space-occupying lesions that obliterated the turbinates, caused deviation of the nasal septum, and eroded bone . Rhinitis appeared as a cavitating lesion that spared the paranasal sinuses, thickened and distorted the turbinates, and widened the meatus . Although morphologically distinct on computed tomographic images, infectious rhinitis and nasal neoplasms could not be differentiated by attenuation measurements or degree of contrast enhancement . Computed tomography appeared to be a reliable, noninvasive technique for the diagnosis of chronic nasal disease in dogs, and a promising alternative to diagnostic techniques currently in use.

J Clin Microbiol, 1993 Apr, 31(4), 831 - 5
Use of DNA analysis of Pasteurella haemolytica biotype T isolates to monitor transmission in bighorn sheep (Ovis canadensis canadensis); Jaworski MD et al.; Pneumonia has been identified as a major cause of poor lamb survival in indigenous herds of Rocky Mountain bighorn sheep (Ovis canadensis canadensis) in central Idaho . Pasteurella haemolytica was isolated from five adult Rocky Mountain bighorn ewes captured from a free-ranging herd in central Idaho . The lambs from two of these ewes delivered by cesarean section were free of P . haemolytica until 40 days of age and after repeated contact with their dams . The lambs subsequently developed signs of pneumonia, and P . haemolytica was isolated from nasal, pharyngeal, and transtracheal wash samples from each lamb . All P . haemolytica biotype T isolates from the ewes and lambs, as well as those from a 9-month-old lamb of the same herd from which samples for culture were obtained 2 years earlier, were subjected to HaeIII restriction enzyme analysis (REA) and ribotyping . Two ribotypes and seven REA patterns were visually distinguishable by these procedures . Similarity coefficients (SAB) of 0.09 to 0.95 were calculated for the seven REA patterns . The REA patterns of the isolates from the lambs were identical (SAB = 1.0) . The isolates from the lambs also had SAB values of 1.0, which was indicative of identity with one of the seven isolates cultured from the ewes at the time of capture and with the organism isolated from the 9-month-old lamb . These procedures have the discriminatory capabilities necessary to monitor the transmission of specific strains of bacteria within and between animal populations.

Avian Dis, 1993 Apr-Jun, 37(2), 616 - 21
Characteristics of fowl cholera diagnosed in Georgia, 1989-91; Waltman WD et al.; One hundred seventy-six cases of fowl cholera were diagnosed at the Georgia Poultry Laboratory over a 3-year period . The disease occurred throughout the year, with peak incidence during March and April . Fowl cholera was diagnosed in flocks from 4 to 83 weeks of age, with a mean of 33 weeks of age . The Pasteurella multocida isolates were highly susceptible to all antimicrobial agents tested, except sulfonamides . The serotypic distribution of isolates showed that serotype 3,4 predominated (40%), followed by serotype 3 (22%), serotype 1 (18%), untypable (15%), serotype 5 (3%), and serotype 4 (2%) . Associations were found between the P . multocida serotypes isolated from birds of different ages and between the age of the bird and the sites of choice for isolating the organism.

Proc Natl Acad Sci U S A, 1993 Mar 15, 90(6), 2495 - 9
Epitopic structure of Tn glycophorin A for an anti-Tn antibody (MLS 128); Nakada H et al.; Glycophorin A was digested with glycoprotease (Pasteurella haemolytica) and the digest was fractionated by a combination of high-pressure column chromatographies to produce the glycopeptides GPA-1 to GPA-6 . Sequence analysis of the glycopeptides revealed that two serine residues (Ser-14 and Ser-15) are not glycosylated, Thr-17 and Ser-19 being glycosylated instead, in disagreement with the accepted structure . The glycopeptides thus obtained were treated with sialidase and beta-galactosidase . The Tn antigenicity, as assayed by the binding to a monoclonal anti-Tn antibody (MLS 128), was found exclusively in the glycopeptides including three (cluster I) or four (cluster II) consecutive residues of GalNAc-Ser/Thr, whereas the glycopeptide (GPA-2) containing two nonconsecutive GalNAc-Ser/Thr residues had practically no Tn antigenicity . The immunoreactivities of GPA-1 and GPA-3, containing both clusters I and II, and GPA-4, containing cluster II, were 63% (calcd . 67%), 81% (calcd . 86%), and 50% (calcd . 50%), respectively, of the immunoreactivity of GPA-5 or GPA-6, containing cluster I (the average being taken as the basis), based on the reactivity per GalNAc residue . These results indicate that clusters I and II react with the antibody to the same extent . The structure consisting of three consecutive glycosylated Ser/Thr residues may be essential for Tn antigenicity in the light of previous results for ovine submaxillary mucin.

Br Vet J, 1993 Mar-Apr, 149(2), 183 - 93
The role of induced virulence factors produced by Pasteurella haemolytica in the pathogenesis of bovine pneumonic pasteurellosis: review and hypotheses; Gonzalez CT et al.; In the pathogenesis of pneumonic pasteurellosis, there is an abrupt commensal to pathogen shift from a predominance of P . haemolytica serotype 2 (ST2) to serotype 1 (ST1) in the bovine upper respiratory tract (URT) microfloral population . This occurs following periods of stress associated with development of this disease . Data are reviewed from recent publications supporting the contention that surface-expressed ST1-specific factor(s) could be critical in mediating URT adhesion and colonization . Such factors may promote an increase in the number of ST1 organisms deposited through infective droplets into the lungs, beyond that efficiently cleared by normal lung defences . The seeding of these organisms into the lungs may provide numerous foci of infection that eventually progress into characteristic pneumonic lesions seen in the disease.

Zentralbl Mikrobiol, 1993 Mar, 148(2), 83 - 7
{The localization of toxin in the cells of Pasteurella multocida}; Erler W; The application of the degradation procedure for Gram-negative bacteria according to Bewick and Lo to Pasteurella multocida indicates that the obvious localization of the toxin is in the periplasm . The stability of the outer membrane and of the substances adhering to it is essential for the release of the toxin . The production of the toxin clearly depend on the media used.

Dtsch Tierarztl Wochenschr, 1993 Mar, 100(3), 99 - 102
{Comparison of EBL cells and ELISA in the culture and serological diagnosis of rhinitis atrophicans in swine}; Alt M et al.; Pasteurella multocida isolates from 271 nasal swabs of pigs were tested in EBL cell culture for toxin production . Mixed bacteria cultures of the same swabs were examined in the P . multocida toxin ELISA K462 (Dakopatts) . In the ELISA 114 swabs reacted positive, whereas toxigenic P . multocida were detected by the EBL cell test in 86 swabs . In a neutralization test (SNT) combined with EBL cell culture and with the ELISA 111 sera were examined for P . multocida antitoxin . The toxin had to be more concentrated for the ELISA than for the cell culture; therefore the SNT with EBL cells was more sensitive . Whereas 101 sera had titres of 1:4 or higher in the cell culture, 68 of these sera were positive in the ELISA.

Berl Munch Tierarztl Wochenschr, 1993 Mar, 106(3), 83 - 4
{The effect of toxins from Pasteurella multocida type D in calves in vivo}; Erler W et al.; By intratracheal injection of purified dermonecrotic toxin from a Pasteurella multocida type D strain, pneumonitis in calves could be produced . This demonstrates the important role of this toxin in the pathogenesis of pneumonitis in calves.

Vet Microbiol, 1993 Mar, 34(3), 287 - 302
Further characterization of Pasteurella haemolytica-like bacteria isolated from swine enteritis; Oberst RD et al.; DNA-DNA hybridization studies were conducted on six Pasteurella haemolytica-like (PHL) organisms recovered from cases of swine enteritis . Chromosomal-enriched fractions of PHL organisms served as the source of DNA for Southern blots or as whole-chromosomal DNA probes . Under stringent hybridization conditions, chromosomal DNA probes of a prototype PHL (strain 6213A) organism distinguished other PHL organisms from Pasteurella haemolytica types A1 and T3, Pasteurella multiocida types A:1 and A:3, Escherichia coli, Pseudomonas aeruginosa, Actinobacillus pleuropneumoniae type 1, and Salmonella cholerasuis . The guanine-cytosine content of the DNA of three PHL strains was 41.2 to 42.8 mol % as calculated from the thermal denaturation midpoint temperatures . The PHL strains are Gram-negative, nonmotile, beta-hemolytic, pleomorphic, oxidase-positive, urease- and indole-negative, fermentative rods with the key characteristics of the species Pasteurella haemolytica . None of the PHL strains reacted with the type-specific antisera of P . haemolytica types 1 through 12 as tested by an agglutination procedure . These swine strains differed in their biochemical differentiation from P . haemolytica types A1 and T3 in that all produced acid from M-inositol and failed to grow on MacConkey agar . Acid production from trehalose and L-arabinose was variable with PHL strains . Leukotoxicity of PHL strains was evaluated by a colorimetric micro-titration assay . Sterile culture supernatants of three of five PHL strains were toxic to bovine neutrophils . Results of these studies suggest that the PHL organisms may belong to a new group of organisms under the genus Pasteurella.

Res Commun Chem Pathol Pharmacol, 1993 Mar, 79(3), 389 - 92
Vaccination with live Pasteurella haemolytica alters vascular adrenergic reactivity; Weekley LB et al.; Rats were injected with sterile saline (controls) or 0.7 x 10(5) c.f.u . of Pasteurella haemolytica (biotype A) obtained from a commercial vaccine . Three days following vaccination animals were killed and aortas removed from the heart base . Isolated aorta was placed in a tissue bath and the biophysical responses to phenylephrine (alpha-1 agonist), clonidine (alpha-2 agonist) and isoproterenol (beta agonist) determined . The results indicate that the alpha-2 adrenergic contractile response to clonidine is significantly enhanced while the response to phenylephrine is not significantly altered . The relaxant response to isoproterenol is eliminated following vaccination . These results suggest that exposure to vaccine-derived strains of P . haemolytica cause an imbalance in the adrenergic responsiveness of vascular smooth muscle.

Rev Sci Tech, 1993 Mar, 12(1), 237 - 72
Report of the Thirteenth Meeting of the OIE Ad hoc Group on Non Tsetse-Transmitted Animal Trypanosomoses; Touratier L; There is increasing interest in many parts of the world in trypanosomoses other than those transmitted by tsetse flies, as shown by numerous research projects and field studies . The refinement of techniques for studying the behaviour of trypanosomes (techniques of molecular biology) in axenic culture or in the parasitised host has led to progress in diagnosis and immunology, and a rational approach to chemotherapy and chemoprophylaxis of these infections . Field trials of enzyme-linked immunosorbent assays in Africa, Asia and South America have shown that these tests may now be regarded as reliable in demonstrating antibodies or antigens for Trypanosoma evansi infection in buffalo, cattle and camels, and for mono-infection with T . equiperdum in equines . However, it is not yet possible to differentiate reliably between infections with T . evansi and T . equiperdum in equines . The card agglutination trypanosomosis test (CATT) has been adapted to T . evansi infection and can also be recommended . Immunosuppression induced by T . evansi infection inhibits the immune response to vaccination against Pasteurella haemolytica . In areas freed from tsetse flies (Cameroon, Central African Republic, Zambia) it has been observed that Trypanosoma vivax can be transmitted mechanically by other biting insects, which are at present being identified . Research on trypanocides has led to the toxic factor for Trypanosoma brucei or T . equiperdum present in human or simian serum being localised to the high density lipoprotein of serum lipoproteins . Various derivatives are being tested under laboratory conditions, and the efficacy of some (e.g . ronidazole) is being checked at present, while others are ready to pass to the development stage (e.g . IMOL 881) . Melarsomine, already available commercially (as Cymelarsan) for the treatment of T . evansi infection in camels, is being studied for possible use in other species of animals.

Carbohydr Res, 1993 Feb 24, 240, 277 - 85
Characterization of the O-polysaccharide of Pasteurella haemolytica serotype A1; Severn WB et al.; Lipopolysaccharide was isolated from Pasteurella haemolytica biotype A, serotype 1 by using the phenol-water extraction procedure . Hydrolysis with mild acid afforded a high-molecular-weight antigenic O-chain . On the basis of 1D and 2D NMR spectral studies and microanalytical chemical methods, the O-polysaccharide was determined to be a linear polymer of a trisaccharide repeating unit having the structure -->3)-beta-D-Galp-(1-->3)-beta-D-GalpNAc-(1-->4)-beta-D-Galp-(1--> This O-polysaccharide antigen is expressed by several P . haemolytica biotype A serotypes.

Nippon Jibiinkoka Gakkai Kaiho, 1993 Feb, 96(2), 192 - 6
{Paranasal sinusitis due to Pasteurella multocida}; Nakano H et al.; A case of paranasal sinusitis due to Pasteurella multocida (P . multocida) is reported . A 39-year-old woman presented with chief complaints of rhinorrhea and headache . The patient kept a cat in her house and kept such close contact with it as to wake up by being licked every morning . Bacteriological examination revealed P . multocida isolated from her nasal discharge and also from the saliva of the cat kept by the patient . The two isolates were compatible with respect to biochemical properties, serotype and drug susceptibility . Therefore, P . multocida infection in this case was considered to have originated from the pet cat . P . multocida infection has been increasing recently . One of the reasons is a pet boom . In order to prevent acquiring the infection from a pet animal, we should have knowledge about this infection, advise the patient to avoid close contact with pets, and provide valuable information concerning these problems to society from the viewpoint of zoonosis.

Vet Microbiol, 1993 Feb, 34(2), 167 - 73
Protection of Pasteurella multocida dermonecrotic toxin-challenged rats by toxoid-induced antibody; Pettit RK et al.; Two different doses of glutaraldehyde-treated Pasteurella multocida dermonecrotic toxin (PMDT) were used to immunize rats . Rats developed serum IgG antibodies specific for native PMDT, and IgG titers increased with dose and number of toxoid immunizations . Survival rates in both active immunization and passive serum neutralization experiments were dependent on dose of toxoid vaccination and serum levels of anti-PMDT IgG . Vaccination with toxoid prevented weight loss but not leukocytosis and increased complement titers in toxin-challenged rats . Toxoid, itself, induced minimal leukocytosis but no alterations in complement titers or weight gain.

J Clin Microbiol, 1993 Feb, 31(2), 364 - 7
Comparison of isolation methods for the recovery of Bordetella bronchiseptica and Pasteurella multocida from the nasal cavities of piglets; Lariviere S et al.; Nasal swabs from 241 piglets from 12 herds with clinical atrophic rhinitis and 283 piglets from 14 herds without clinical atrophic rhinitis were examined for the presence of Bordetella bronchiseptica and/or Pasteurella multocida . For B . bronchiseptica, swabs were streaked on three selective media . Blood agar supplemented with cephalexin was the most satisfactory selective culture medium for the isolation of B . bronchiseptica . For P . multocida, swabs were also streaked on three selective media . Mice were also used for isolation of P . multocida from the nasal cavities of pigs . The mouse inoculation test was not found to be the definitive test for the isolation of P . multocida . A significant number of P . multocida strains were avirulent in the mouse model . The modified Knight medium (without potassium tellurite) was the best single method for isolating P . multocida . However, a combination of mouse passage and direct culture on selective media increased the rate of isolation . There was no marked difference in the prevalence of B . bronchiseptica or P . multocida in swine herds with or without clinical atrophic rhinitis . Both capsular types A and D were present in the nasal cavities of the pigs with or without clinical atrophic rhinitis.

J Clin Microbiol, 1993 Feb, 31(2), 255 - 9
Comparison of DNA fingerprinting and serotyping for identification of avian Pasteurella multocida isolates; Wilson MA et al.; The DNA fingerprint profiles and serotypes of 63 avian Pasteurella multocida field isolates, 13 attenuated vaccine isolates (propagated from vaccines manufactured by five companies), and 16 somatic reference strains were compared . DNA fingerprinting established the relationship of isolates that could not be distinguished by serotyping . Of the 76 isolates, 28 DNA fingerprint profiles and 12 somatic types were recognized . One isolate was nonreactive with 16 reference somatic and 5 reference capsule-type antisera . Thirty-one field isolates and seven vaccine isolates were identified as capsule type A . Twenty-nine field isolates and six vaccine isolates were nonencapsulated . Three field isolates were capsule type F . Isolates of capsule types B, D, and E were not found . One field isolate, identified as somatic type 7, had a DNA fingerprint identical to that of the somatic reference type 6 profile . Twelve field isolates had profiles identical to the somatic reference type 3 strain profile; 11 of these were identified as somatic type 3, 4, and 1 was identified as somatic type 3 . The DNA fingerprint profiles of 50 field isolates and 13 attenuated vaccine isolates did not match profiles of the 16 somatic type reference strains . Twenty-five DNA fingerprint profiles were recognized from 30 of these field isolates . The DNA fingerprint profiles of 20 field isolates and 13 attenuated vaccine isolates were identical . Three somatic types (3; 3,4; and 4,16) were identified from the field isolates, and two somatic types (3 and 3,4) were identified from the attenuated vaccine isolates . DNA fingerprinting is useful for accurate identification and epidemiologic study of P . multocida isolates.

Zentralbl Hyg Umweltmed, 1993 Feb, 194(1-2), 214 - 22
{Pets as permanent excretors of zoonoses pathogens}; Mayr B; When scrutinizing zoonoses with regard to risks for human beings, the spectrum of pathogens with dogs, cats and birds leading to persistent infections and consequently to the fact that the animals become carriers and permanent excretors is relatively small . Most of the zoonoses cause clinical symptoms and will be taken care of correspondingly . With regard to dogs there is a multitude of persistent infections that are transferred from the pet to the human being and vice versa . In reality, however, the importance of the dog as permanent excretor of zoonosis pathogens endangering human health is minimal, except for some parasitoses . As far as cats are concerned, the situation is totally different . Cats are carriers and permanent excretors of pasteurella, the pathogens of the so-called cat-scratch disease, trichophyton and microsporum species, toxoplasmosis and orthopox viruses . The new zoonosis feline pox serves as an example of the necessity of a permanent observation of persistently infected pets . Healthy, but persistently infected birds form a source of infection not to be underestimated . Through the beat of their wings they constantly stir up dried infectious excrements and dust and thus favour the airborn infection of human beings . Chlamydia psittaci, the Newcastle disease virus and Mycobacterium avium are of major importance in this context . The risk of transferring zoonosis pathogens from persistently infected pets to human beings can be minimized through prophylactic diagnosis, strict measures of hygiene, observation of the schedule of vaccinations for the respective species and regular use of anthelmintica.

Microbiol Immunol, 1993, 37(2), 103 - 9
Drug resistance and broad geographical distribution of identical R plasmids of Pasteurella piscicida isolated from cultured yellowtail in Japan; Kim EH et al.; An MIC test of 12 chemotherapeutic agents performed on 175 strains of Pasteurella piscicida collected from cultured yellowtail (Seriola quinqueradiata) in different areas of Japan from 1989 to 1991 revealed 152 strains (87%) with resistance to combinations of ampicillin (AP), chloramphenicol (CP), kanamycin (KM), nalidixic acid (NA), sulfamonomethoxine (SA), tetracycline (TC), and/or trimethoprim (TMP) . The remaining 23 strains were sensitive to all the drugs tested: AP, cefazolin, CP, florfenicol (FF), furazolidone, KM, NA, novobiocin, SA, streptomycin, TC, and TMP . FF showed the most effective antibacterial activity against P . piscicida with MICs ranging from 0.004 to 0.6 microgram/ml . One hundred and forty-nine of the 152 resistant strains carried transferable R plasmids encoding one of the Cp Km Sa Tc, Km Sa Tc, Km Sa, and Sa resistance . The most common resistance marker of transferable R plasmids identified in P . piscicida was Km Sa Tc . R plasmids encoding three different resistant markers were very similar on the basis of their digestion patterns with restriction endonucleases . There was homology among the DNAs of nine transferable R plasmids selected . Our findings suggest that multiple drug resistant strains of P . piscicida carrying transferable R plasmids with the same DNA structure are common in yellowtail farms and that the R plasmid has been retained within the P . piscicida population without change in their DNA structure according to geography and year.

Vet Surg, 1993 Jan-Feb, 22(1), 27 - 30
Bacterial isolates from blood cultures of dogs undergoing dentistry; Harari J et al.; Bacteria in blood cultures in 30 dogs undergoing high-speed dental scaling and tooth extraction were examined . One or more positive blood cultures were identified in 9 of 30 (30%) dogs . Pasteurella spp . were most frequently (5 dogs) isolated and were sensitive to ampicillin, penicillin, cephalothin, chloramphenicol, tetracycline, amoxicillin with clavulanic acid, and sulfamethoxazole with trimethoprim . Two groups of 15 dogs each, anesthetized or sedated but not undergoing dental procedures, served as non-dentistry controls . There were no significant (p < .05) differences between the number of positive cultures in dentistry and non-dentistry groups . In healthy dogs undergoing high-speed dental scaling and tooth extraction, the occurrence of bacteria in blood cultures was much lower than previously reported . The clinical significance of positive blood cultures was uncertain.

J Comp Pathol, 1993 Jan, 108(1), 81 - 91
Effects of Pasteurella multocida toxin on the osteoclast population of the rat; Martineau-Doize B et al.; Pasteurella multocida type D toxin is a peptide shown to induce severe atrophic rhinitis in the pig as the result of an increased osteoclastic resorption of the ventral nasal turbinates . In the present study, the effects of the toxin on the histological, cytochemical and ultrastructural features of the osteoclast population of the rat were examined . Pasteurella multocida toxin induced atrophy of the ventral and dorsal nasal turbinates and thinning of the nasal bones . The number and size of the long bone metaphyseal osteoclasts were significantly increased, but not the number of nuclei per cell . Osteoclasts of toxin-treated rats had more developed clear zones and ruffled borders than those of the controls and their cytoplasmic vacuoles were more abundant and larger . We concluded that P . multocida toxin stimulates bone resorption by osteoclasts in the rat by increasing resorption activity and by increasing their number . Its action is not limited to the nasal turbinates but occurs also in the other bones, such as the long bones.

J Comp Pathol, 1993 Jan, 108(1), 65 - 72
Morphological and functional disturbances in pulmonary vascular endothelium following exposure of sheep to an "avirulent" strain of Pasteurella haemolytica; Weekley LB et al.; Sheep were vaccinated with a live attenuated strain of Pasteurella haemolytica and killed 3 days later . Segments of main intrapulmonic artery and vein were removed for biophysical and scanning electron microscopic studies . In the pulmonary artery, vaccination with Pasteurella haemolytica caused an increase in the number of endothelial cell surface blebs and, in some cases, those blebs appeared to be splitting open, suggesting cell damage or irritation . There was a surprising lack of platelet adherence to the lesions, suggesting that an antiplatelet factor is released by the damaged endothelium . The endothelial-dependent relaxant response to bradykinin was enhanced following vaccination . In the pulmonary vein, ultrastructural lesions similar to those in the artery were present in vaccinated animals . Bradykinin caused a contraction, an effect that was reduced following vaccination with Pasteurella haemolytica . These experiments demonstrate that a live, vaccine-derived strain of Pasteurella haemolytica causes both morphological and functional changes in the pulmonary vascular endothelium.

Avian Dis, 1993 Jan-Mar, 37(1), 6 - 9
Classification, pathogenicity, and drug susceptibility of hemolytic gram-negative bacteria isolated from sick or dead chickens; Lin MY et al.; Fifteen hemolytic gram-negative bacteria were isolated from the respiratory tracts of sick birds suffering from a long-lasting respiratory syndrome or from the bone marrow of dead birds distributed in the southern part of Taiwan . These were classified as Pseudomonas aeruginosa (10 isolates), Pseudomonas fluorescens (2 isolates), Pseudomonas stutzeri (1 isolate), Pasteurella haemolytica (1 isolate), and Proteus morganii (1 isolate) . Each isolate was inoculated intraperitoneally into one group of ten 4-week-old male white leghorn chickens . Mortality and lesions were scored daily for 1 week . Three of the 10 isolates of Pseudomonas aeruginosa caused 100% mortality . Six other isolates of Pseudomonas aeruginosa and the one isolate of Proteus morganii caused 50% mortality . The remaining isolates induced less than 30% mortality . The sole nonpathogenic sample was one isolate of Pseudomonas fluorescens . When therapeutic levels of 22 antibiotics or sulfa drugs were evaluated for their inhibitory activity against the 15 isolates, the most effective were apramycin (15/15), gentamicin (15/15), spectinomycin (13/15), oxytetracycline (8/15), and sulfachloropyrazine (7/15) . The least effective were ampicillin, cloxacillin, and tiamulin, which were not effective against any of the isolates . The 14 other drugs were of very low (> 4/15) effectiveness . Most of the isolates studied were virulent for chickens and very resistant to currently used drugs.

J Wildl Dis, 1993 Jan, 29(1), 30 - 5
Pasteurella haemolytica cytotoxin-dependent killing of neutrophils from bighorn and domestic sheep; Silflow RM et al.; Peripheral blood neutrophils from Rocky Mountain bighorn sheep (Ovis canadensis canadensis) and domestic sheep were exposed to culture supernatants from Pasteurella haemolytica isolates recovered from these two sheep species . Six culture supernatants from bighorn sheep isolates and two from domestic sheep isolates were tested for cytotoxicity as determined by the release of lactate dehydrogenase . Two of the bacterial culture supernatants from bighorn sheep were not cytotoxic, while the other four bighorn sheep culture supernatants were effective cytotoxins on both bighorn (> 95% cell death at 150 micrograms of cytotoxin) and domestic sheep neutrophils (55 to 95% cell death at 150 micrograms of cytotoxin) . Two culture supernatants of P . haemolytica from domestic sheep were effective cytotoxins on both bighorn (> 95% cell death at 150 micrograms of cytotoxin) and domestic sheep (70 to 75% cell death at 150 micrograms of cytotoxin) neutrophils . Potency of cytotoxins derived from P . haemolytica isolates from bighorn sheep was three to seven-fold higher when tested with bighorn sheep neutrophils as compared to domestic sheep neutrophils . Cytotoxins derived from P . haemolytica isolates from domestic sheep were five to six-fold more potent when tested with bighorn sheep neutrophils than when domestic sheep cells were used.

Histol Histopathol, 1993 Jan, 8(1), 97 - 104
Experimental reproduction of acute pneumonic pasteurellosis in rabbits; Redondo E et al.; A histomorphometric and physiopathological study was made of the lung parenchyma of Belgian White SPF rabbits infected experimentally with Pasteurella multocida type A . Symptoms observed were characteristic of the acute respiratory syndrome . Mean serum cortisol concentration and rectal temperature increased in all experimental groups . Histopathological changes included alveolitis and leukocytic bronchitis . Changes in alveolar and bronchial cytoarchitecture were attributed to the degeneration and necrosis of constituent epithelial cells.

Comp Immunol Microbiol Infect Dis, 1993 Jan, 16(1), 77 - 85
Characterization of Pasteurella from gingival scrapings of dogs and cats; Ganiere JP et al.; Gingival scrapings of 62 dogs and cats were examined for the presence of Pasteurella . Isolation was performed in a medium supplemented with thiostrepton . Twenty-eight and 37 strains were obtained from 21 dogs and 26 cats, respectively, and classified in recently described species or subspecies of the genus Pasteurella (P.): P . multocida subspecies multocida and septica, P . canis, P . dagmatis and P . stomatis . Twenty-one strains were classified as atypical P . stomatis and one strain obtained from a cat remained unclassified . All strains were susceptible to the antibiotics studied . P . multocida and P . stomatis (including atypical strains) represented 65 and 30% of feline isolates, and 14 and 68% of canine isolates, respectively . Assuming that P . multocida, P . canis and P . dagmatis are potentially pathogenic for humans, and that P . stomatis has a low pathogenicity or non-pathogenic, 77 and 28% of examined cats and dogs harboured one or several pathogenic strains . This difference could explain the fact that Pasteurella infections in man are lower in dog bites rather than cat bites.

Res Vet Sci, 1993 Jan, 54(1), 20 - 4
Capsular hyaluronic acid in Pasteurella multocida type A and its counterpart in type D; Pandit KK et al.; Hyaluronic acid was demonstrated in the capsule extract of 39/39 Pasteurella multocida type A strains by sodium chloride gradient chromatography followed by Alcian blue staining, and by a turbidometric method using acidified horse serum . Treatment with hyaluronidase from various sources eliminated these reactions . An Alcian blue staining substance of closely similar chromatographic properties occurred in capsule extracts of 14/16 type D strains but it resisted hyaluronidase and was thought to be an acidic polysaccharide differing from the hyaluronic acid of type A . Turbidometric values were lower than with type A strains, but were unaltered by hyaluronidase treatment . The type D substance could be precipitated from capsule extract by acriflavin . Both type A and D strains were mucoid and displayed large capsule zones in stained preparations . Photomicrographic measurements showed that hyaluronidase treatment of cell suspensions markedly reduced capsule dimensions of type A but not type D strains . When type A strains were cross streaked against a hyaluronidase + Staphylococcus aureus, their growth became non-mucoid at the intersection: mucoid type D strains were unaffected . Neither hyaluronic acid nor the hyaluronidase resistant type D substance could be detected in type B or E strains.

Vet Immunol Immunopathol, 1993 Jan, 35(3-4), 353 - 64
Serum levels of tumor necrosis factor-alpha in calves experimentally infected with Pasteurella haemolytica A1; Pace LW et al.; The purpose of this study was to document the levels of tumor necrosis factor-alpha (TNF) in serum of calves experimentally infected intratracheally with Pasteurella haemolytica A1 and to determine if elevated TNF levels correlate with development of pneumonic pasteurellosis in the bovine . Serum samples were collected at sequential time periods from 0 h to 72 h post inoculation with P . haemolytica . TNF levels in those sera were measured by a cytotoxicity assay utilizing the TNF-sensitive WEHI 164 mouse fibrosarcoma cell line . Serum TNF levels in infected cattle began to rise at 2 h post inoculation, peaked at approximately 8 h, and decreased to near control levels by 72 h . There was extreme variability in serum TNF among the inoculated animals with levels varying from 120 pg ml-1 to 5000 pg ml-1 at 8 h post inoculation . These levels did not correspond with the degree of lung involvement . All inoculated calves developed lesions of pneumonic pasteurellosis characterized by fibrinous pleuritis with necrotizing, hemorrhagic pneumonia . These results suggest that TNF is probably a significant inflammatory mediator involved in the pathogenesis of bovine pneumonic pasteurellosis.

Am J Vet Res, 1993 Jan, 54(1), 92 - 8
Identification of Pasteurella haemolytica A1 isolates from market-stressed feeder calves by use of enzyme and antimicrobial susceptibility profiles; Purdy CW et al.; An epidemiologic study of Pasteurella haemolytica serovar 1 (Ph1) in market-stressed feeder calves from 7 farms in eastern Tennessee was conducted . The nasal mucus of each calf was cultured sequentially at the farm of origin (day 0), at an auction market (day 133), and at a feedyard in Texas (days 141, 148, 155, and 169) . Of the 103 calves tested, 77 were culture-positive, including 1 on day 0, 1 on day 133, 20 on day 141, 57 on day 148, 50 on day 155, and 14 on day 169 . From the 143 Ph1 isolates, 20 enzyme profiles were determined by use of a commercial enzyme system that detects 19 enzymatic reactions; 4 antimicrobial susceptibility profiles were obtained, using the disk-diffusion method, which evaluated susceptibility to 11 antibacterial drugs . All isolates were positive for acid phosphatase and alkaline phosphatase, but were negative for alpha-galactosidase, alpha-mannosidase, beta-glucosidase, beta-glucuronidase, cystine aminopeptidase, N-acetyl-beta-glucosaminidase, and trypsin . Other positive enzyme reactions included: leucine aminopeptidase, 140 Ph1 isolates; phosphohydrolase, 90 isolates; alpha-fucosidase, 63 isolates; esterase (C4), 59 isolates; valine aminopeptidase, 30 isolates; esterase lipase (C8), 24 isolates; beta-galactosidase, 2 isolates; and alpha-glucosidase, chymotrypsin and lipase (C14), 1 isolate each . Thirty-four Ph1 profiles were identified, using combined enzyme and antimicrobial susceptibility profiles . The data indicate that the strains isolated during the feedyard period may have been determined more by farm of origin (P < or = 0.001) than by habitation with calves from other farms while in the feedyard.(ABSTRACT TRUNCATED AT 250 WORDS)

Infect Immun, 1993 Jan, 61(1), 253 - 9
Neuraminidase production by a Pasteurella haemolytica A1 strain associated with bovine pneumonia; Straus DC et al.; The properties of an extracellular neuroaminidase produced by a Pasteurella haemolytica A1 strain (isolated from a case of bovine pneumonia) during growth in a defined medium were examined in this investigation . This enzyme, isolated from concentrated culture supernatants of P . haemolytica A1, was active against N-acetylneuramin lactose, human alpha 1-acid glycoprotein, fetuin, and bovine submaxillary mucin . Neuraminidase production paralleled bacterial growth in a defined medium and was maximal in the stationary phase of growth . The enzyme was purified to homogeneity by a combination of salt fractionation, ion-exchange chromatography on DEAE-Sephacel, and gel filtration on Sephadex G-200 . These procedures yielded an enzyme preparation that possessed a specific activity of 100.62 mumol of sialic acid released per min per mg of protein against human alpha 1-acid glycoprotein . The Km value for this enzyme with human alpha 1-acid glycoprotein as the substrate was 1.1 mg/ml, and the enzyme possessed a pH optimum of 6.5 . The P . haemolytica A1 neuraminidase had a molecular weight of approximately 150,000 as estimated by gel filtration and approximately 170,000 when analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis . The enzyme was stable at 4 degrees C for 3 h . At 37 degrees C for 3 h, 25% of enzymatic activity was lost . Approximately 55% of the enzyme activity was lost within 30 min at 50 degrees C, with greater than 70% of the enzyme activity being destroyed within 10 min at temperatures of > or = 65 degrees C.

Infect Immun, 1993 Jan, 61(1), 170 - 81
Structural and serological specificities of Pasteurella haemolytica lipopolysaccharides; Lacroix RP et al.; Lipopolysaccharides (LPSs) from 16 serotypes of Pasteurella haemolytica were subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis and examined by silver staining and immunoblotting . Silver staining of proteinase K-digested cell lysates revealed two rough LPS serotypes (serotypes 2 and 8), which lacked demonstrable O-polysaccharide, while 14 others demonstrated a ladder pattern characteristic of smooth-type LPS . Purified LPSs from several serotypes yielded O-polysaccharide in addition to low-molecular-weight core oligosaccharide components when subjected to mild acid hydrolysis . Nuclear magnetic resonance spectroscopy revealed the O-chain polysaccharides of serotypes 1, 6, and 9 to be identical . Immunoblots using hyperimmune rabbit, mouse, bovine, and ovine sera from homologous and heterologous serotypes supported this finding and suggested that most of the A biotypes share common O-chain epitopes . Immunoblotting results also supported structural data which demonstrated that the O-polysaccharides of serotypes 3 and 15 and of serotypes 4 and 10 (T biotypes) are identical . Nuclear magnetic resonance analysis indicated that the core oligosaccharides of serotypes 1, 6, 8, 9, and 12 share similar structures, but that they are distinct from those of serotypes 3, 4, 10, and 15 . Immunoblots with hyperimmune antisera and monoclonal antibody having specificity for the core region of serotype 1 LPS revealed shared epitopes in the core oligosaccharides of several A biotypes . Characterization of the molecular structure and antigenic specificities of LPS has been an important consideration in the development of purity and potency assays for veterinary vaccines which contain P . haemolytica.

Chest, 1993 Jan, 103(1), 7 - 11
Pasteurella multocida pneumonia in a man with AIDS and nontraumatic feline exposure; Drabick JJ et al.; A case of acute pneumonia due to Pasteurella multocida ssp multocida occurred in a young man with AIDS and chronic sinusitis . The pneumonia was diagnosed by bronchoscopy and responded to treatment with aztreonam . Epidemiologic investigation revealed the case was temporally related to nontraumatic exposure to cat secretions that the patient presumably had acquired via an aerosol . The cat's oral cavity was cultured and an isolate of P multocida ssp multocida with identical biochemical reactions, DNA restriction patterns, and nearly identical fatty acid profile to that of the patient's isolate was obtained suggesting they were identical strains and therefore epidemiologically linked . A control strain with identical biochemical reactions and antibiotic sensitivities exhibited different patterns . To our knowledge, this is the first such reported infection in a patient infected with human immunodeficiency virus.

Acta Vet Scand, 1993, 34(1), 29 - 34
Aerobic bacteria occurring in the vagina of bitches with reproductive disorders; Bjurstrom L; A retrospective survey was performed of aerobic bacterial species found in the vagina of 203 bitches with genital disorders, e.g . infertility, vaginitis, pyometra and puppy death . Escherichia coli, beta-hemolytic streptococci, Staphylococcus intermedius and Pasteurella multocida were the species most often isolated . From bitches with pyometra E . coli in pure culture was the most frequent isolate . In contrast, the majority of infertile bitches gave rise to mixed cultures, and no specific bacterial species was consistently associated with infertility . Thus, bacterial sampling from infertile bitches was concluded to be of low diagnostic value . Bacterial species isolated from the bitches having vaginitis were present in pure culture in 26.9% of the samples while nonspecific mixed cultures were obtained from 34.6% of the samples from these bitches . E . coli was the most frequently isolated bacterial species from bitches with dead puppies . However, in such cases it is important to relate the vaginal bacterial findings to autopsy findings and the results of bacteriological cultures of the pups.

Vaccine, 1993, 11(7), 767 - 72
Clinical and serological evaluation of a Pasteurella haemolytica A1 capsular polysaccharide vaccine; Conlon JA et al.; The purified capsular polysaccharide (CPS) of Pasteurella haemolytica A1 was examined for its ability to protect cattle from experimental challenge with logarithmic-phase P . haemolytica . Several preparations of P . haemolytica antigens were utilized in the experiment including CPS, log-phase P . haemolytica culture supernatant, P . haemolytica recombinant leucotoxin (rLKt) and various combinations of the above . CPS alone or in combination with culture supernatant or rLkt elicited no protection; rather, administration of CPS was associated with a high incidence of anaphylaxis (36% of calves) . Although a classical biphasic humoral immune response to CPS could be detected in all calves that received this compound, this T-dependent response was not correlated with resistance to experimental challenge . The complexity of protective immunity in pneumonic pasteurellosis is emphasized by this study, and clinical anaphylaxis associated with response to CPS may be implicated in the pathogenesis of disease.

Scand J Infect Dis, 1993, 25(5), 655 - 8
Pasteurella multocida meningitis in a two-day old neonate; Hillery S et al.; A normal full-term baby boy, born by vaginal delivery, became ill on day 2 with fever and failure to feed . CSF examination revealed 260 x 10(6)/l leucocytes, mainly mononuclears, protein 2 g/l and glucose zero . Pasteurella multocida was isolated in pure culture from the baby's CSF, blood and umbilicus and from the mother's vagina . The baby was treated with i.v . penicillin for 7 weeks . Progress was complicated by mild hydrocephalus, which resolved, and prolonged low grade fever . Recovery was complete, without neurological sequelae . This case illustrates that P . multocida can infect the vagina where it presents a hazard to a newborn infant delivered vaginally . Early diagnosis is critical, intravenous high dose penicillin being the treatment of choice.

DNA Seq, 1993, 3(6), 357 - 67
Cloning, sequencing and expression of a Pasteurella haemolytica A1 gene encoding a PurK-like protein; Chang YF et al.; A membrane protein antigen of Pasteurella haemolytica A1 encoded on the recombinant plasmid pYFC13 is isolated and characterized . Nucleotide sequence analysis of the insert DNA in pYFC13 identified the gene mpa1, which codes a protein of approximately 45 kDa without signal sequence . The deduced amino acids from the DNA sequence are homologous to Bacillus subtilis PurK by 29.4%; to Schizosaccharomyces pombe Pur6 by 29.34%, to Saccharomyces cerevisiae Pur6 by 25.867%; and to E . coli PurK by 25.223% identity, respectively . The purK and pur6 from these organisms are responsible for the activity of 5'-phosphoribosyl- 5-amino-4-imidazole carboxylase which is involved in de novo purine biosynthesis . The protein was over-expressed in E . coli by its own promoter . The antigen we designated as Mpa1, could be localized to the cytoplasmic membrane of both P . haemolytica A1 and E . coli TB1 harbored pYFC13 . The Mpa1 was antigenic in rabbit and in cattle since both animals produced antibody against this protein.

Vet Res Commun, 1993, 17(2), 143 - 51
Oscillatory measurements, blood gas analysis and clinical observations after intravenous clenbuterol administration in healthy and acutely pneumonic calves; Reinhold P; The effects of clenbuterol (Ventipulmin, Boehringer Ingelheim) on respiratory functions were investigated in 6 calves aged 4-6 weeks prior to and after experimental infection with Pasteurella haemolytica A1 . On days 1-3 (prior to infection) and on days 7-9 (after infection), blood gas analysis, monofrequency forced oscillation techniques and clinical examinations (heart rate, respiratory rate) were conducted for 135 min after the intravenous administration of clenbuterol (0.8 microgram/kg body weight) . In healthy calves prior to Pasteurella infection, intravenous administration of clenbuterol induced a mild tachycardia and a reduction in the mean oscillatory respiratory resistance . Using the same dose of clenbuterol in diseased calves after infection, the statistically significant reduction in oscillatory respiratory resistance was more impressive and it was accompanied by a significant increase in the oxygen pressure of the arterialized blood . Heart rate and respiratory rate did not change significantly after the administration of clenbuterol in infected calves.

FEMS Microbiol Lett, 1992 Dec 15, 79(1-3), 125 - 31
An analysis of the codon usage of Pasteurella haemolytica A1; Lo RY; Analysis of approximately 17 kbp of nucleotide sequences from three different regions of the genome of Pasteurella haemolytica A1 showed that the mol% G+C of P . haemolytica A1 DNA is 38.5% . When only the coding sequences (approx . 10 kbp) were analysed, a similar value of 38.8% was obtained . A comparison of the relative synonymous codon usage values of the cloned genes showed that P . haemolytica A1 has a very different codon usage pattern from that of Escherichia coli.

J Biol Chem, 1992 Dec 15, 267(35), 25239 - 45
Interconversion of GRP78/BiP . A novel event in the action of Pasteurella multocida toxin, bombesin, and platelet-derived growth factor; Staddon JM et al.; Incubation of Swiss 3T3 cells with {2-3H}adenine, as in other cell types, reveals the ADP-ribosylation of GRP78 (the 78-kDa glucose-regulated protein, also known as BiP, the immunoglobulin heavy chain-binding protein), a resident endoplasmic reticulum protein that assists in the processing of proteins destined for secretion or cell surface expression . Here we show that Pasteurella multocida toxin, a potent growth factor for cultured fibroblasts, decreased the ADP-ribosylation of GRP78/BiP to 16 +/- 6% of the control value (n = 23) . The action of the toxin occurred after a lag period, was blocked by lysosomotrophic agents, and potentiated by increased incubation time (ED50 4 ng/ml and 1 ng/ml in 4 and 8 h, respectively), thus indicating that the toxin enters the cells to act . Bombesin and platelet-derived growth factor (PDGF) similarly decreased the ADP-ribosylation of GRP78/BiP (ED50 0.5 nM and 2.5 ng/ml, respectively) but acted more rapidly than the toxin . Signaling pathways activated by the toxin, bombesin, and PDGF had effects on the ADP-ribosylation of GRP78/BiP . Thus, activation of protein kinase C alone by phorbol 12,13-dibutyrate was partially effective, and down-regulation of protein kinase C attenuated but did not block the action of the toxin, bombesin, and PDGF . Agents that mobilize Ca2+ from the endoplasmic reticulum (A23187, ionomycin, and thapsigargin) caused a decrease in the ADP-ribosylation of GRP78/BiP that was similar in magnitude to that achieved by the toxin, bombesin, and PDGF, implicating a role for inositol 1,4,5-trisphosphate-mediated Ca2+ mobilization in the action of the mitogenic agents . The growth factor-induced decrease in the ADP-ribosylation of GRP78/BiP may represent its conversion from an inactive to an active state.

J Biol Chem, 1992 Dec 15, 267(35), 25296 - 303
Pasteurella multocida toxin selectively facilitates phosphatidylinositol 4,5-bisphosphate hydrolysis by bombesin, vasopressin, and endothelin . Requirement for a functional G protein; Murphy AC et al.; Treatment of Swiss 3T3 cells with a subsaturating concentration of recombinant Pasteurella multocida toxin (rPMT) markedly potentiated the production of inositol phosphates induced by bombesin, vasopressin, and endothelin but not by platelet-derived growth factor (PDGF) (AA and BB homodimers) . Similarly, the neuropeptides but not PDGF caused a shift in the dose-dependent increase in inositol phosphates induced by rPMT . The rate of accumulation of inositol phosphates induced by bombesin was increased 2-fold by rPMT treatment while that of PDGF was unaffected . rPMT treatment also enhanced bombesin-induced inositol(1,4,5)trisphosphate, the direct product of phosphatidylinositol 4,5-bisphosphate hydrolysis . In contrast, treatment of cells with rPMT had no effect on the tyrosine phosphorylation of phospholipase C gamma . Depletion of protein kinase C increased rPMT-induced inositol phosphates in a manner similar to that observed for bombesin but not PDGF . Thus, rPMT selectively potentiates neuropeptide-mediated inositol phosphate production . The action of rPMT on phosphatidylinositol 4,5-bisphosphate hydrolysis persisted in streptolysin O-permeabilized cells . Addition of guanosine 5'-O-(beta-thiodiphosphate) to permeabilized cells markedly reduced rPMT-induced inositol phosphates in a time- and dose-dependent manner . rPMT also increased the sensitivity of phospholipase C for free calcium . Our results strongly suggest that the action of rPMT facilitates the coupling of G protein to phospholipase C.

Vet Rec, 1992 Dec 5, 131(23), 528 - 31
Some aspects of the epidemiology and control of Salmonella typhimurium infection in outwintered suckler cows; Davies TG et al.; Two outbreaks of Salmonella typhimurium infections affected outwintered, spring-calving suckler cows in late pregnancy . The infections spread rapidly both within and between groups of stock on the affected farms, with morbidity in the infected groups varying from 14.5 per cent to over 60 per cent, and mortality in adult cattle varying from 0 to 14.3 per cent . Prophylactic measures included the use of antibiotics and killed vaccines against Escherichia coli, Salmonella dublin, S typhimurium, and Pasteurella multocida . In one outbreak, use was also made of a polyvalent serovaccine and hyperimmune serum against E coli, S typhimurium, and S dublin . In both outbreaks no new cases were reported in the affected groups after the administration of the second dose of vaccine, and there was no resurgence of disease on the affected farms within 18 months of the primary outbreaks.

J Gen Microbiol, 1992 Dec, 138 ( Pt 12), 2491 - 8
Pathogenic activities of live cells and extracellular products of the fish pathogen Pasteurella piscicida; Magarinos B et al.; The pathobiological activities in vivo and in vitro of live cells and extracellular products (ECP) of eleven Pasteurella piscicida strains of different origin were examined . Infectivity trials showed that P . piscicida did not possess strict host specificity since the majority of the isolates were virulent for gilthead seabream, rainbow trout and turbot, with LD50 values ranging between 10(3) and 10(6) live cells . However, none of the strains tested were pathogenic for mice (LD50 > 10(8) cells)) . In addition, the ECP were strongly toxic for fish (LD50 ranging from 1.0 to 4.6 micrograms protein per g fish), which clearly demonstrates their important role in the pathogenesis of pasteurellosis . All the ECP samples were cytotoxic for fish and homoiothermic cell lines, possessed notable phospholipase activity and displayed haemolytic activity for sheep, salmon and turbot erythrocytes (but not for trout erythrocytes) . However, the production of proteolytic enzymes differed among the P . piscicida strains . Although no strain displayed elastase activity, five isolates (the Japanese and Italian strains) hydrolysed casein and gelatin . All these biological activities in vivo and in vitro were lost after heat treatment (100 degrees C for 10 min) . The general enzymic patterns of both live cells and ECP evaluated by the API-ZYM system also revealed some variation among the P . piscicida isolates . Generally, whole cells showed a wider range of enzymic activities than ECP . The results presented here are important for the selection of strains in the development of effective polyvalent pasteurellosis vaccines containing both whole cells and ECP.

J Vet Med Sci, 1992 Dec, 54(6), 1225 - 7
The first outbreak of fowl cholera in Muscovy ducks (Cairina moschata) in Japan; Nakamine M et al.; The first outbreak of fowl cholera occurred in a flock of Muscovy ducks (Cairina moschata) in Okinawa Prefecture of Japan in November 1990 . Fifty (25%) of 200 birds in a farm died of an acute disease . Remaining birds recovered after treatment with oxytetracycline . Pasteurella multocida subsp . multocida was isolated in pure culture from all tissues tested from two dead birds . Serovars of the isolates were identified as Carter's capsular type A . Heddleston's type 3.4.12, and Namioka's type 5:A which have not been demonstrated in Japan . Pathologically, multiple necrosis and bacterial aggregates were prominent in several organs, particularly in the liver . The isolate killed chickens when inoculated intravenously at a concentration of 10(8) colony forming units.

J Vet Med Sci, 1992 Dec, 54(6), 1105 - 10
Interaction between Bordetella bronchiseptica and toxigenic Pasteurella multocida on the nasal mucosa of SPF piglets; Elias B et al.; The interaction between Bordetella bronchiseptica and type D toxigenic Pasteurella multocida was studied in five groups of 4 specific-pathogen-free (SPF) piglets each . At 28 days of age, piglets of groups 3 and 4 were inoculated into both nostrils with 10(8) colony-forming-units (CFU) of a non-dermonecrotic toxin (DNT)-producing, phase I strain of B . bronchiseptica . Piglets of groups 1 and 3 were treated intranasally with a sonic extract of the non-toxic strain of B . bronchiseptica and those of groups 2 and 4 with B . bronchiseptica DNT into the left nostril . Sonic extract and DNT treatment was started at 33 days of age and lasted for 5 days . Piglets of group 5 served as controls . At the age of 37 days, piglets of all groups except group 5 were inoculated into both nostrils with 5 x 10(7) CFU of toxigenic P . multocida . At slaughter at 50 days of age, P . multocida was recovered from the left nasal cavity of 3 piglets of group 2 and all piglets of group 4 . In piglets inoculated with B . bronchiseptica DNT the mucosal epithelial cells of the left nasal cavity showed loss of cilia, regressive lesions such as vacuolation, karyopycnosis and necrosis, hypertrophy of the epithelium, infiltration of the epithelium and submucosa by inflammatory cells, could also be seen . The results suggest that action of the B . bronchiseptica DNT on the nasal mucosa is a precondition of the growth of P . multocida in the nasal cavity.(ABSTRACT TRUNCATED AT 250 WORDS)

Appl Environ Microbiol, 1992 Dec, 58(12), 4072 - 5
Application of ozone disinfection to remove Enterococcus seriolicida, Pasteurella piscicida, and Vibrio anguillarum from seawater; Sugita H et al.; Survival of bacterial fish pathogens, including Enterococcus seriolicida, Vibrio anguillarum, and Pasteurella piscicida, in ozonated seawater was determined in a batch system . Bacterial counts of all fish pathogens decreased at more than 0.040 to 0.060 mg of total residual oxidants (TROs) per liter, whereas no decrease in viable counts was observed at less than 0.018 to 0.028 mg of TROs per liter . The 99% inactivation point was achieved at concentrations of 0.111 mg/liter for E . seriolicida, 0.063 mg/liter for P . piscicida, and 0.064 mg/liter for V . anguillarum within 1 min . Moreover, the mean 99 and 99.9% killing concentration-contact time (C.t) products were 0.123 and 0.186 mg.min/liter for E . seriolicida, 0.056 and 0.084 mg.min/liter for P . piscicida, and 0.081 and 0.123 mg.min/liter for V . anguillarum, respectively . However, the mean 99 and 99.9% C.t products for the mixed population in coastal seawater were 0.200 and 0.621 mg.min/liter . These results strongly suggest that ozone treatment at more than 1.0 mg of TROs per liter for several minutes is able to disinfect seawater for mariculture efficiently.

Mol Microbiol, 1992 Dec, 6(23), 3585 - 93
Molecular analysis of the aroA gene of Pasteurella multocida and vaccine potential of a constructed aroA mutant; Homchampa P et al.; The aroA gene from Pasteurella multocida was cloned by complementation of the Escherichia coli aroA mutant AB2829 with a DNA library constructed in pUC18 . The nucleotide sequence of the P . multocida aroA gene indicated an open reading frame encoding a protein of 441 amino acids, which showed a high degree of homology with the amino acid sequences of various other bacterial AroA proteins . The cloned P . multocida aroA gene was inactivated by insertion of a kanamycin-resistance gene and reintroduced by allelic exchange into the chromosome of P . multocida using the suicide vector pJM703.1 . The P . multocida aroA mutant was highly attenuated in a mouse model . Mice immunized intraperitoneally with two doses of live P . multocida aroA mutant were completely protected against a lethal parental strain challenge.

Am J Clin Pathol, 1992 Dec, 98(6), 565 - 8
Pasteurella multocida endocarditis; Hombal SM et al.; Human infection with Pasteurella multocida is the leading cause of animal bite wound infection . Life-threatening infection may occur in patients with a variety of underlying disorders and an immunocompromised state . Infective endocarditis with P . multocida is very rare and only a few clinically diagnosed cases have been reported . Described here is an autopsy case of a 61-year-old man with polycystic kidney disease who had P . multocida bacteremia and acute infective endocarditis with multiple bacterial clumps involving bicuspid aortic valve . The organisms were gram negative . Apparently the sepsis with P . multocida was acquired via licking of leg ulcers by his pet dog, establishing an animal-related causal relationship . Because P . multocida is a very common flora of many animals, infection with this organism probably occurs more frequently than is commonly appreciated . High index of suspicion and early diagnosis, especially in immunocompromised patients, are warranted because the disease is potentially life threatening, yet is a readily treatable infection.

Infect Immun, 1992 Dec, 60(12), 5182 - 9
Ovine pulmonary surfactant induces killing of Pasteurella haemolytica, Escherichia coli, and Klebsiella pneumoniae by normal serum; Brogden KA; Pulmonary surfactant has been shown to play an increasingly important role in bacterial clearance at the alveolar surface in the lung . This study describes a bactericidal mechanism in which ovine pulmonary surfactant induces killing of Pasteurella haemolytica by normal serum . To demonstrate killing, six bacterial species were incubated first with pulmonary surfactant for 60 min at 37 degrees C and then with serum for an additional 60 min at 37 degrees C . P . haemolytica type A1 strains 82-25 and L101, a P . haemolytica type 2 strain, Escherichia coli, and Klebsiella pneumoniae were susceptible and Pasteurella multocida, Serratia marcescens, and Pseudomonas aeruginosa were not susceptible to killing by ovine pulmonary surfactant and normal serum . No bacteria incubated with bovine pulmonary surfactant were killed by normal serum . Although the species origin of pulmonary surfactant was selective, the species origin of serum was not . P . haemolytica incubated with ovine pulmonary surfactant was killed by fetal calf serum, gnotobiotic calf serum, pooled normal sheep serum, pooled normal rabbit serum, and pooled guinea pig serum . Ultrastructurally, killed P . haemolytica suspensions contained dead cells and cells distorted with vacuoles between the cytoplasmic membrane and the cytoplasm . The mechanism of killing did not correlate with concentrations of complement or lysozyme or titers of residual antibody in either the pulmonary surfactant or the serum, and killing was reduced by preincubation of surfactant with P . haemolytica lipopolysaccharide . Preliminary characterization of both surfactant and serum implicate a low-molecular-weight proteinaceous component in the surfactant and serum albumin in the serum . This mechanism may help clear certain gram-negative bacteria from the lungs of sheep as a part of the pulmonary innate defense system.

Infect Immun, 1992 Dec, 60(12), 4984 - 8
Effect of Pasteurella multocida toxin on bone resorption in vitro; Felix R et al.; Pasteurella multocida toxin (PMT), which is the primary etiologic factor in the pathogenesis of progressive atrophic rhinitis in pigs, was found to stimulate bone resorption in vitro . This stimulation was observed both in cultures of murine calvaria by measuring the release of calcium and of the lysosomal enzyme beta-glucuronidase and in murine long bone cultures by measuring the release of calcium . Both systems showed the same dose response curve, with the maximal effect at a concentration of 5 ng/ml . The effect on calvaria was studied in more detail . PMT increased bone resorption 24 h after its addition and always had to be present to express an effect . Calcitonin was able to inhibit this increase of resorption completely, and inhibitors of prostaglandin synthesis suppressed it partially . Although the data show an effect of PMT on bone tissue, the results do not exclude an action on cells in the nasal cavity, which could indirectly stimulate bone resorption.

Rev Sci Tech, 1992 Dec, 11(4), 1163 - 8
Serological survey for bovine bacterial and viral pathogens in captive Arabian oryx (Oryx leucoryx Pallas, 1776); Greth A et al.; Tests for antibodies to bovine bacterial and viral pathogens were conducted on 239 sera from 128 Arabian oryx (Oryx leucoryx) from seven locations (Taif, Riyadh and Mahazat as Said, Saudi Arabia; San Diego, United States of America {USA}; Shaumari, Jordan; Qatar; and Bahrain) . No antibodies to Pasteurella multocida type E or epizootic haemorrhagic disease 1 virus were found . Antibodies to Brucella abortus, P . multocida type B, P . multocida type D, lumpy skin disease virus and Akabane virus were detected in 2, 1, 5, 2 and 1 animals, respectively . Evidence of P . multocida type A, Coxiella burnetti, Chlamydia psittaci and parainfluenza 3 virus was found in 3 herds (prevalence in the main herd {n = 78}: 8%), 3 herds (8%), 6 herds (7%) and 5 herds (15%), respectively . Evidence of antibodies against bluetongue virus was found in five oryx from the USA and in one oryx from the Taif herd . Antibody vaccinal titres against rinderpest virus (and the virus of peste des petits ruminants, due to cross-reactions) were found in almost all the herds . This is the first report of antibodies against B . abortus, C . burnetti, C . psittaci, parainfluenza 3 virus and Akabane virus in the genus Oryx.

Isr J Med Sci, 1992 Dec, 28(12), 847 - 51
Leishmania major: bacterial contamination of cutaneous lesions in experimental animals; el-On J et al.; No bacterial contamination has been demonstrated in cutaneous leishmaniasis (CL) nodule and in lesions caused by Leishmania major in Balb/c mice up to 20 days after infection . However, although many phagocytic cells (polymorphonuclear leukocytes and macrophages) were present in the CL lesion, 80% of the lesions showed bacterial contamination that developed within the first 70 days of infection . Topical treatment of the lesion with an ointment containing 15% paromomycin and 12% methylbenzethonium chloride in soft white paraffin for 20 days eliminated all the Leishmania parasites and several of the associated bacteria including: Proteus vulgaris, Pasteurella multocida, Staphylococcus albus and Staphylococcus aureus . This treatment did not affect Escherichia coli, Klebsiella spp . and Pseudomonas aeruginosa . Total elimination of these bacteria was achieved only during the healing process, and within 20 days following termination of treatment . The rate of disappearance of bacteria inoculated alone into the base of the tail of normal uninfected Balb/c mice was much faster than that of bacteria inoculated into either the CL nodule or the CL lesion . This study suggests the development of local immunosuppression in the CL lesion that may be mediated by the Leishmania parasites and their metabolites.

Zentralbl Veterinarmed B, 1992 Nov, 39(9), 649 - 61
Polymyxin B: pharmacokinetics of single doses given intravenously and intramuscularly to turkeys, and minimal inhibitory concentrations for Escherichia coli and Pasteurella multocida; Freidlin PJ et al.; The 50% and 90% minimal inhibitory concentrations (MIC50 and MIC90) of polymyxin B for avian Escherichia coli and Pasteurella multocida isolates were determined by the agar plate dilution method . Polymyxin B at approximate MIC level in serum was bactericidal for E . coli in 2 to 4 hours . Aqueous polymyxin B sulfate was administered by a single bolus intravenous injection into turkeys at 10,000 IU/kg, and by a single bolus intramuscular injection at 5,000, 10,000 or 20,000 IU/kg . Effective serum drug concentrations after intramuscular injection (MIC50 levels or greater) were maintained for E . coli for 7.0 hr (10,000 IU/kg) and 11.5 hr (20,000 IU/kg), and for P . multocida for 3.0 hr (10,000 IU/kg) and 4.1 hr (20,000 IU/kg) . Pharmacokinetic parameters were calculated by non-compartmental methods . Elimination time half-lives, mean residence time, clearance, and apparent volume of distribution at steady state (Vdss) were all much higher for i.m . injection of 20,000 IU/kg than for i.m . injection of 10,000 IU/kg . We postulate that there exists a minimal tissue-interaction threshold concentration (MTC) at which polymyxin B can enter previously unavailable compartments or bind to previously refractory tissue components . Bioavailability of polymyxin B injected i.m . was 0.904 for the 10,000 IU/kg dose and 0.675 for the 20,000 IU/kg dose . Dosage intervals necessary to produce minimal steady state concentrations (Cssmin) equal to the MIC were calculated . Certain aspects of the use of the parameter Vdss, and limitations on the use of dosage interval calculations for polymyxin B, are discussed . One week after i.m . injection of polymyxin B at 10,000 IU/kg, high tissue drug levels were present, especially in bound form in liver . Following single injections, no toxic effects on turkeys were observed.

Berl Munch Tierarztl Wochenschr, 1992 Nov 1, 105(11), 378 - 80
{Possibilities for the establishment of pneumonia-free swine herds by immunization and administration of cefquinome}; Schimmel D; The application of Bordetella and Pasteurella inactivated and adsorbed vaccines together with cefquinome to sows, piglets and weaners led to a significant reduction of the incidence of rhinitis atrophicans and pneumonia . The frequency of positive isolates of P . multocida, H . parasuis and A . pleuropneumoniae out of nasal swabs was reduced during the treatment.

J Clin Microbiol, 1992 Nov, 30(11), 2984 - 7
Characterization and distribution of Pasteurella species recovered from infected humans; Holst E et al.; During a 3-year period, all Pasteurella strains recovered at the Clinical Microbiological Laboratory, Lund, Sweden, were studied biochemically with respect to their relationship to the recently described taxa of this genus . Of 159 strains recovered from 146 infected humans, 95 were identified as Pasteurella multocida subsp . multocida, 21 as Pasteurella multocida subsp . septica, 28 as Pasteurella canis, 10 as Pasteurella stomatis, and 5 as Pasteurella dagmatis . The homology within and between the Pasteurella species regarding cellular fatty acids and enzymatic activities was also studied . Strains of the different Pasteurella species were indistinguishable from each other regarding fatty acid composition; all strains contained major amounts of C14:0, C16:1, C16:0, and 3-OH-C14:0 acids and minor amounts of C18:2, C18:1, and C18:0 acids . Neither did the enzymatic activities distinguish between strains belonging to different species . In addition, of 56 strains examined, toxin production was demonstrated only in 1 strain each of P . multocida subsp . multocida and P . canis . Except for one severe case of necrotizing cellulitis involving P . dagmatis, P . multocida subsp . multocida or P . multocida subsp . septica was recovered in the more serious cases of infection . Except for P . canis, which in all cases was associated with dog bites, most Pasteurella strains were recovered in cases of infection associated with cat bites or scratches . Pasteurella strains occurred in four infected patients without evident connections with animals.

J Leukoc Biol, 1992 Nov, 52(5), 558 - 64
Activation of bovine neutrophils by Pasteurella haemolytica leukotoxin is calcium dependent; Ortiz-Carranza O et al.; In this study, we used the fluorescent probe Fluo-3 to show that an increase in cytosolic free calcium, {Ca2+}i, occurred when suspensions of bovine neutrophils were incubated with sublethal concentrations of P . haemolytica leukotoxin . This increase in {Ca2+}i was dependent on the concentration of leukotoxin present in the medium and, at a given concentration of leukotoxin, dependent on the external calcium concentration . The calcium channel blocker verapamil and the beta-adrenergic antagonist propranolol inhibited leukotoxin-stimulated Ca2+ gain, as did a neutralizing antileukotoxin monoclonal antibody . As reported previously, incubation of bovine neutrophils with partially purified leukotoxin stimulated a vigorous luminol-dependent chemiluminescence response (LDCL) . The present study shows that LDCL stimulation was dependent on the presence of extracellular calcium and was inhibited by the addition of verapamil and propranolol . These data indicate that bovine neutrophils exhibit a considerable increase in cytoplasmic free calcium when they are incubated with P . haemolytica leukotoxin in the presence of external calcium . They also provide evidence that an increased {Ca2+}i is required for functional activation of the bovine neutrophil oxidative burst by P . haemolytica leukotoxin.

Biochem Biophys Res Commun, 1992 Oct 30, 188(2), 760 - 6
The glycoprotease of Pasteurella haemolytica A1 eliminates binding of myeloid cells to P-selectin but not to E-selectin; Steininger CN et al.; HL-60 cells and neutrophils treated with the glycoprotease from Pasteurella haemolytica A1, an enzyme which is specific for O-sialoglycoproteins, were found to be incapable of binding P-selectin but still bound E-selectin . Comparative analysis of {35-S} cysteine labeled proteins from HL-60 cells by 2-dimensional electrophoresis indicated that two major proteins with M(r) 100 and 115 kd were significantly removed from cells which had been treated.

Avian Dis, 1992 Oct-Dec, 36(4), 986 - 91
Turkey macrophage and heterophil bactericidal activity against Pasteurella multocida; Harmon BG et al.; Bactericidal activity of turkey macrophages and heterophils was demonstrated in an in vitro colorimetric bactericidal assay . Two vaccine strains and one field isolate of Pasteurella multocida A:3,4 and a single isolate each of Escherichia coli and Staphylococcus aureus were compared for susceptibility to the bactericidal activity of turkey macrophages and heterophils . Only P . multocida A:3,4-strain M-9 (the least virulent strain) was susceptible to macrophage bactericidal activity in the absence of specific immune serum, whereas all three P . multocida A:3,4 organisms were killed when opsonized with specific immune serum . E . coli was susceptible to the bactericidal activity of macrophages, and S . aureus was resistant . All bacteria tested were highly sensitive to the bactericidal activity of intact turkey heterophils, regardless of the opsonin treatment . Electron microscopic findings suggested that heterophils may kill extracellular P . multocida . Only S . aureus and E . coli were killed by lysed heterophils.

Avian Dis, 1992 Oct-Dec, 36(4), 975 - 85
Vaccination of turkeys with cell-free culture filtrate of Pasteurella multocida: effects of dilution, iron chelation, and heterologous challenge; Ficken MD et al.; Two experiments were done to further define cell-free culture filtrate (CCF) from Pasteurella multocida and its endotoxin content in protecting turkeys against challenge . In the first experiment, the greater-than-30,000-molecular-weight fraction of P . multocida strain R44/6 (serotype 3/4/9/12) CCF was used in 10-fold dilutions given by air-sac inoculation or aerosol to vaccinate turkeys, which were subsequently challenged with either homologous (P-1059, serotype 3) or heterologous (X-73, serotype 1) strains . Endotoxin content of the CCF fraction was high . Compared with positive controls given either live Clemson University vaccine or a commercial bacterin, homologous protection was provided by undiluted CCF and 1:10 dilutions of CCF, but there was no heterologous protection . In the second experiment, CCF of strain R44/6 in regular and iron-limiting media and CCF of strain FC127B (serotype 1/4) were used alone or in combination to vaccinate turkeys, which were challenged as in the first experiment . Homologous but not heterologous protection occurred, even though growth of strain R44/6 in iron-limiting media reduced endotoxin content of CCF by approximately 93% . These results indicate that endotoxin levels of less than 10% but greater than 1% of those in CCF from regular media are sufficient to induce protection in turkeys against homologous challenge but that CCF from either regular or iron-limiting medium does not provide protection against heterologous challenge.

Cell Mol Neurobiol, 1992 Oct, 12(5), 499 - 510
The macrophage-activating properties of growth hormone; Edwards CK 3rd et al.; 1 . We compared the ability of growth hormone (GH) and a well-characterized macrophage-activating factor, interferon-gamma (IFN-gamma) to activate highly purified populations of alveolar macrophages . Both GH and IFN-gamma primed macrophages triggered with opsonized zymosan to secrete superoxide anion (O2-) in vitro, but IFN-gamma was effective at a 40-fold lower concentration . Antibody blocking studies demonstrated that the priming activity of GH was independent of IFN-gamma, and the activity of IFN-gamma was distinct from that of GH . 2 . Both IFN-gamma and GH increased the capability of macrophages to kill Pasteurella multocida in vitro . 3 . Hypophysectomized rats challenged with Salmonella typhimurium were significantly protected by injections of either GH or recombinant rat IFN-gamma in vivo compared to vehicle-treated controls, and the protective effect of GH was increased by incorporation into liposomes . 4 . Insulin-like growth factor-I (IGF-I) also primed alveolar macrophages in vitro, which is consistent with the idea that the protective effects of GH in vivo might be mediated by augmenting the synthesis of IGF-I . These data support the concept of reciprocal systems of communication between the neuroendocrine and immune systems.

J Vet Diagn Invest, 1992 Oct, 4(4), 419 - 22
Use of ELISA to detect toxigenic Pasteurella multocida in atrophic rhinitis in swine; Bowersock TL et al.; The use of an enzyme-linked immunosorbent assay (ELISA) as a means of detecting dermonecrotoxin-producing strains of Pasteurella multocida was investigated . The assay was evaluated as a means to identify toxigenic P . multocida isolates recovered from nasal secretions of swine with atrophic rhinitis . The sensitivity and specificity of the ELISA for detecting dermonecrotoxin-producing P . multocida strains were compared to those of mouse-inoculation and cytotoxicity assays . The ELISA was highly sensitive and more specific than animal inoculation or tissue culture assay and is thus a more effective method for screening swine herds for the presence of toxigenic strains of P . multocida . The ELISA is a rapid, effective, economical way to identify toxigenic P . multocida isolates.

Am J Vet Res, 1992 Oct, 53(10), 1889 - 94
Systemic and pulmonary antibody responses of calves to Pasteurella haemolytica after intrapulmonary inoculation; McBride JW et al.; Systemic and pulmonary antibody responses of calves to Pasteurella haemolytica were evaluated by measuring immunoglobulin production in blood for 9 days and in pulmonary lavage fluid for 7 days after intrapulmonary inoculation . Clinical signs, pulmonary lesions, pulmonary and systemic inflammatory response, and amount of antigen in lavage fluid were used to evaluate the response of calves to challenge with P haemolytica . The pulmonary response consisted of production of IgG, IgE, and IgM antibodies to P haemolytica antigens and a 17- to 68-fold increase of cells in lavage fluid 8 hours after inoculation, with a gradual decrease toward normal . Antibodies of the IgM isotype to P haemolytica were demonstrated as early as 8 hours through 7 days after inoculation in 3 of 3 calves . Of the anti-P haemolytica isotypes, IgM was found in the highest concentration . In all of the inoculated calves, IgE was found 1 to 2 days after inoculation, and IgG was found in 2 of 3 inoculated calves from day 1 through 7 after inoculation . Detection of IgG correlated with smaller pulmonary lesions . Immunoglobulin A was not detected in lavage fluid . Serum was evaluated for IgG and IgM antibody response to P haemolytica . Specific IgM was detectable 5 days after inoculation, and IgG was detectable 7 days after inoculation . Pasteurella haemolytica antigens were not detected in serum or plasma . A transient increase in neutrophil count was found 8 hours after inoculation, with return to baseline values by 24 hours after inoculation.(ABSTRACT TRUNCATED AT 250 WORDS)

Appl Environ Microbiol, 1992 Oct, 58(10), 3316 - 22
Phenotypic, antigenic, and molecular characterization of Pasteurella piscicida strains isolated from fish; Magarinos B et al.; We compared Pasteurella piscicida strains isolated from different fish species in several European countries with strains isolated in Japan and the United States . The taxonomic analysis revealed that, regardless of the geographic origin and source of isolation, all the strains exhibited the same biochemical and physiological characteristics . Serological assays with different rabbit antisera demonstrated a high level of antigenic similarity among strains, with cross-agglutination titers of 20,480 to 40,960 . This serological homogeneity was supported by the lipopolysaccharide (LPS) and membrane protein profiles . All the P . piscicida strains had the same electrophoretic LPS pattern, showing O side chains with a ladder-like structure, and shared at least four major outer membrane proteins, of 20, 30, 42, and 53 kDa . Western blot (immunoblot) analysis with LPS and protein indicated that all the P . piscicida strains are immunologically related . In addition, the chromosomal DNA fingerprint patterns obtained for the European strains with the enzymes EcoRI and BamHI were practically identical to those of the Japanese and U.S . strains . Although some differences were found in the plasmid profiles of P . piscicida, a large number of strains possessed in common plasmid bands of 20 and 7 MDa . In addition, a plasmid of 50 MDa was present in the majority of the European strains . Restriction endonuclease analysis demonstrated the genetic homology of the plasmid bands shared by most of the European strains . All the P . piscicida strains had the same drug resistance patterns, indicating that a correlation between plasmid carriage and resistance to a specific antimicrobial agent cannot be established . The high levels of phenotypic, serological, and genetic homogeneity found among the P . piscicida strains should facilitate the development of DNA probes with diagnostic purposes as well as the design of effective vaccines.

Antimicrob Agents Chemother, 1992 Oct, 36(10), 2093 - 8
Effects of sub-MICs of antibiotics on cell surface characteristics and virulence of Pasteurella multocida; Lebrun A et al.; The effects of sub-MICs of certain antibiotics, namely, penicillin G, tetracycline, and trimethoprim-sulfamethoxazole, on the cell surface characteristics and the virulences of two toxigenic isolates of Pasteurella multocida representing capsular types A and D were evaluated . Expression of proteins, in particular, outer membrane proteins and iron-regulated proteins, was not affected by exposure of bacterial cells to low concentrations of antibiotics . However, exposition of surface antigens was modified by sub-MICs of the antibiotics tested . The lipopolysaccharide profile of one isolate (capsular type D) was altered by penicillin G . Sub-MICs of penicillin G and tetracycline diminished the virulence of the capsular type A isolate and adherence to porcine tracheal rings of the capsular type D isolate . Production of dermonecrotic toxin was not affected by sub-MICs of the antibiotics tested . Our results indicate that growth of P . multocida in the presence of low concentrations of antibiotics seems to have, depending on the isolate, profound effects on cell surface characteristics, with concomitant effects on adherence or virulence . Our results also indicate that production of dermonecrotic toxin, an important virulence factor of P . multocida isolates associated with porcine atrophic rhinitis, was not affected by sub-MICs of the antibiotics studied.

Avian Dis, 1992 Oct-Dec, 36(4), 964 - 7
In vitro susceptibility of avian Escherichia coli and Pasteurella multocida to danofloxacin and five other antimicrobials; Raemdonck DL et al.; The in vitro susceptibility of Escherichia coli and Pasteurella multocida isolated from poultry was determined to danofloxacin, a novel fluoroquinolone, and five other commonly used antimicrobials . A total of 1737 E . coli field isolates and 107 P . multocida isolates were tested by veterinary diagnostic laboratories in Europe, Japan, South Africa, and North America during the period 1989-91 . The antimicrobial susceptibility of these isolates was determined using the Sensititre broth microdilution technique . The minimum inhibitory concentrations (MIC) of danofloxacin, furaltadone, lincomycin, oxytetracycline, spectinomycin, and trimethoprim:sulfamethoxazole that prevented growth of 90% of the bacteria were 0.25 > 64, > 64, > 64, > 128, and > 16 micrograms/ml, respectively, against E . coli isolates and 0.25, 64, 64, 16, 128, and 8 micrograms/ml, respectively, against P . multocida isolates . Danofloxacin demonstrated considerable in vitro potency against these important poultry pathogens, many of which showed extensive resistance to the other antimicrobials tested.

Can J Vet Res, 1992 Oct, 56(4), 281 - 8
Serological titers to bovine herpesvirus 1, bovine viral diarrhea virus, parainfluenza 3 virus, bovine respiratory syncytial virus and Pasteurella haemolytica in feedlot calves with respiratory disease: associations with bacteriological and pulmonary cytological variables; Allen JW et al.; Acute and convalescent serum samples were taken from 59 calves with signs of respiratory disease (cases) and 60 clinically normal animals (controls) during their first month in the feedlot . Sera were analyzed for antibodies to bovine parainfluenza 3 (PI3) virus by hemagglutination inhibition, to bovine viral diarrhea (BVD) virus, bovine respiratory syncytial (BRS) virus and bovine herpesvirus 1 (BHV1) by virus neutralization, and to Pasteurella haemolytica by indirect agglutination (PhIA) and cytotoxin neutralization (PhCN) tests . There was minimal evidence of serological activity to BHV1 . Serological activity to the other agents occurred commonly and the prevalence of acute titers and their mean values was similar in case and control groups . Mean convalescent PI3 and P . haemolytica (PhIA) titers were higher in controls than cases (p < 0.01) but, otherwise, convalescent titers did not differ between groups . The incidence of seroconversion was similar in both groups for all agents except for PI3 virus which was more frequent in controls than cases (p < 0.0001) . There was a positive association between PhIA and CN seroconversion and isolation of P . haemolytica from bronchoalveolar lavage (BAL) fluid (p < 0.1) . The measure of agreement (kappa) between seroconversion with the P . haemolytica PhIA and PhCN tests was 0.51 . Bacteriological and cytological evaluations of the respiratory tract were made using BAL . No associations were evident between serological titers and pulmonary cytology . A multivariate logistic analysis was used to evaluate associations between disease status and serological, bacteriological and cytological data . Cases were positively associated with the presence of neutrophils and Pasteurella multocida in BAL fluid and negatively associated with PI3 virus and PhIA seroconversion.

Trop Geogr Med, 1992 Oct, 44(4), 359 - 61
Pasteurella multocida infection in Singapore; Tay L et al.; A case of Pasteurella multocida infection in Singapore is presented . The patient was a 21-year-old Chinese male who developed fever and cellulitis with abscess formation of his right index finger after it was bitten by a stray cat . The organism was isolated in pure culture and identified as Pasteurella multocida subspecies septica . The patient responded to antibiotic therapy and had an uneventful recovery.

J Gen Microbiol, 1992 Oct, 138 ( Pt 10), 2185 - 95
Lipopolysaccharide heterogeneity in Pasteurella haemolytica isolates from cattle and sheep; Ali Q et al.; Lipopolysaccharide (LPS) from 40 isolates of Pasteurella haemolytica, comprising 23 serotype A1, seven serotype A2, one serotype T4, one serotype T10 and eight untypable isolates, obtained from diseased and healthy cattle or sheep, was characterized by SDS-PAGE and Western blotting . Ten different SDS-PAGE LPS profiles, five smooth and five rough, were identified among the biotype A and untypable isolates and designated LPS types 1-10 . LPS types 1 and 2 were smooth, had similar O-antigen banding-patterns but differed in the low-molecular-mass or core-oligosaccharide regions; type 3 LPS was rough but had a core-oligosaccharide region similar to that of LPS type 1 . No similarities were observed between these LPS types and types 6, 7 and 9, which were smooth, and types 4, 5, 8 and 10, which were rough . Most serotype A1 isolates (19/23) were of LPS type 1, whereas two isolates each had LPS of types 2 and 3 . The majority (5/7) of serotype A2 isolates possessed type 3 LPS, whereas the remaining two isolates each had LPS of types 4 and 5 . There was much greater heterogeneity within the untypable group of isolates, which comprised LPS of types 1 and 9 (two isolates each), and 6, 7, 8 or 10 (one isolate each) . Western blotting analysis demonstrated that LPS types 1 and 2 had immunologically identical O-antigen side-chains but differed in their core-oligosaccharide regions, whereas the core-oligosaccharide region of rough LPS type 3 was immunologically very similar to that of LPS type 1 . The other LPS types were immunologically unrelated to these three LPS types . The majority (20/23) of serotype A1 isolates originated from cattle and possessed LPS types 1 or 2, different from most (5/7) of the serotype A2 isolates which originated from sheep and possessed LPS of types 3 or 4 . However, two of the three ovine serotype A1 isolates had the same type 3 LPS as occurred in most of the ovine serotype A2 isolates, suggesting a possible correlation between LPS type and host specificity . This study has demonstrated that LPS diversity within different serotypes of P . haemolytica is greater than was previously thought and that certain LPS types might be host-specific.

Vet Microbiol, 1992 Oct, 32(3-4), 327 - 42
A characterization of monoclonal antibodies prepared against Pasteurella haemolytica serotype 1 surface antigens; Austin FW et al.; The production and characterization of monoclonal antibodies against Pasteurella haemolytica serotype 1 is described . Ten monoclonal antibodies were produced and divided, on the basis of their properties, into six different groups . One produced bacteria agglutination only of P . haemolytica serotype 1 . Three antibodies bound with P . haemolytica serotypes 1, 5-8 and 12 and the antigen was identified in immunoblots as lipopolysaccharide . Two antibodies bound P . haemolytica serotypes 1, 2, 5-8 and 12 and P . multocida serotypes 1-7, 9, 12, 15 and 16, recognizing an epitope present on a 29 kDa outer membrane protein . One antibody bound all P . haemolytica and P . multocida serotypes . The antigen was a hexosamine less than 30 kDa which contained a formalin sensitive epitope . One antibody bound only to P . haemolytica serotype 1 and the antigen was identified as a 66 kDa outer membrane protein . Two antibodies bound P . haemolytica serotypes 1, 2, 5-9 and 12 and the antigen, while not identified, was localized on the outer membrane . This study identified antigens which contribute to the cross-reactions among P . haemolytica and P . multocida serotypes and the antibodies may be useful in investigating the pathogenesis of pneumonic pasteurellosis.

FEMS Microbiol Immunol, 1992 Sep, 5(1-3), 29 - 36
The synthesis and function of the Escherichia coli hemolysin and related RTX exotoxins; Welch RA et al.; The RTX group of exotoxins represents a branch of a family of exoproteins produced by Gram-negative bacteria which share the properties of being secreted by a leader-independent pathway and a tandemly-repeated nine-amino-acid sequence that is responsible for calcium binding . The Escherichia coli hemolysin (HlyA) is the prototype for the RTX exotoxin family which includes the leukotoxins of Pasteurella haemolytica and Actinobacillus actinomycetemcomitans and hemolysins from four Gram-negative genera . A review of the genetics, synthesis, export and target cell reactivity of the E . coli hemolysin is given . An evolutionary tree of the RTX toxin family based on amino acid sequence similarity is presented.

Southeast Asian J Trop Med Public Health, 1992 Sep, 23(3), 520 - 5
Characteristics of Pasteurella multocida isolated from humans, swine and poultry in Thailand; Unchitti K et al.; Pasteurella multocida is a pathogen of animals and humans . Most of the patients have been associated with animals but many cases had not contacted them . The failure to diagnose P . multocida infections is mostly due to misidentification on gram stained smears and inadequate laboratory identification techniques . In order to compile detailed characteristics of the organism we studied the physical and biochemical properties of 70 isolates of P . multocida - 17 human, 23 swine and 30 poultry . All isolates produced catalase, oxydase, indol, nitrate reduction and ornithine decarboxylase . They failed to produce urease, gelatinase, methyl red, acetoin and could not grow on MacConkey agar, SS-agar, in nutrient broth with 0% or 6% NaCl . With respect to fermentable sugars, all isolates consistantly produced acid from glucose, mannitol and mannose . None of the cultures fermented lactose, maltose and dulcitol . Marked variations in the patterns of fermentation of arabinose and xylose were found . The characteristics tested are important to facilitate identification of P . multocida but could not be used to differentiate the host of the bacterium.

Rev Sci Tech, 1992 Sep, 11(3), 917 - 23
Serological evaluation of Pasteurella multocida antigens associated with protection in buffalo calves; Afzal M et al.; Different antigens of Pasteurella multocida Carter's type 6:B including whole bacterium, antigen heated at 56 degrees C, antigen heated at 100 degrees C, sonicated antigen, capsular antigen, potassium thiocyanate extract, lipopolysaccharide and sodium salicylate extract were evaluated to assess protection in buffalo calves against haemorrhagic septicaemia . Sera from calves with known protection status in experimental challenge were titrated by enzyme-linked immunosorbent assay (ELISA) against all antigens . Capsular antigen extracted with 2.5% sodium chloride was superior to other antigens for assessing protection status of buffalo calves against P . multocida by ELISA . This capsular antigen was able to differentiate clearly between well-protected, protected and unprotected animals.

J Arthroplasty, 1992 Sep, 7(3), 309 - 10
Pasteurella multocida infection of a total hip arthroplasty . A case report; Braithwaite BD et al.; The authors report a case history of a diabetic woman requiring revision hip arthroplasty of a Charnley total hip prosthesis that was infected with Pasteurella multocida . The infection of the loose prosthesis followed a cat bite to the same leg . Advice is given on the management of patients with infection following animal inoculations, and the subject of increased risk with a loose prosthesis is discussed.

Leukemia, 1992 Sep, 6(9), 926 - 34
Retention of progenitor cell function in CD34+ cells purified using a novel O-sialoglycoprotease; Marsh JC et al.; We previously showed that the sialoglycoprotein, CD34, which is expressed on primitive human hematopoietic progenitor cells, is cleaved by a unique glycoprotease from Pasteurella haemolytica (P.h . glycoprotease) . This proteolytic enzyme specifically cleaves glycoproteins rich in O-sialoglycans . Glycoproteins containing only N-linked glycans are not cleaved . Cleavage of the CD34 antigen results in the loss of epitopes detected by five of seven CD34-designated antibodies . In this study, we investigated the role of the P.h . glycoprotease in isolating CD34+ cells from unfractionated normal human bone marrow mononuclear cells (MNCs), and determined the effect of the glycoprotease on the proliferative capacity of the progenitor-enriched fraction . CD34+ cells were isolated from MNCs using immunomagnetic beads attached via a CD34 antibody whose epitope is susceptible to removal by the cleavage with the glycoprotease . Subsequent cleavage with P.h . glycoprotease for 30 min at 37 degrees C released the CD34+ cells from the beads with a recovery of up to 78% . Using a CD34 antibody whose epitope was not removed by the glycoprotease, up to 95% of the recovered cells expressed CD34 . Compared to unseparated MNCs, the CD34+ cells showed the following enrichment of committed hematopoietic progenitors, as assayed in semi-solid media: CFU-GM, 45-fold; CFU-M, 13-fold; BFU-E, 26-fold and CFU-GEMM, 81-fold . Hematopoiesis was also studied in two-stage long-term bone marrow cultures in which the CD34+ cells were co-cultured over irradiated, allogeneic adherent layers . Output of CFU-GM over a seven week period from these cultures was similar to that from control cultures with autologous adherent-cell-depleted marrow MNCs . These data suggest that the loss of O-sialo-glycosylated peptide moieties from P.h . glycoprotease-released CD34+ cells neither affects the functional capacity of committed progenitors, nor impairs the proliferation of long-term culture-generating cells . The P.h . glycoprotease can be used to facilitate the isolation and recovery of functionally competent CD34+ cells at high yield and purity, without prior removal of other adherent cells . The ability to rapidly purify CD34+ cells using this non-cytotoxic enzyme has important implications for bone marrow transplantation as well as for gene transfer studies in vitro.

J Med Primatol, 1992 Sep-Oct, 21(7-8), 387 - 8
Pasteurella haemolytica infection in a Goeldie's monkey; Gozalo A et al.; An adult male Callimico goeldii died spontaneously . At necropsy, small whitish foci were found randomly distributed on the liver surface . Histologically, the foci were composed of mixed inflammatory cells with predominant polymorphonuclear cell infiltration and central areas of necrosis . Microbiological cultures revealed a Gram-negative coccoid-bacilli with bipolar staining . Biochemical analysis revealed that the microorganism was Pasteurella haemolytica.

J Med Microbiol, 1992 Aug, 37(2), 128 - 32
Cleavage of immunoglobulin A1, A2 and G by proteases from clinical isolates of Pasteurella multocida; Pouedras P et al.; Several Pasteurella multocida strains were examined for their ability to produce extracellular enzymes that cleave immunoglobulin A and G (Ig A and Ig G) molecules . Two strains isolated from human pulmonary and genital infections produced proteases that cleaved human IgA and IgG, colostral IgA and human myeloma IgA1 and IgA2 . Human IgM was not degraded by these enzymes . Examination of cleavage digests showed two main fragments with different electrophoretic mobilities . The two P . multocida strains produced a protease that cleaved IgA and IgG heavy chains outside the hinge region, and differed in this respect from the hinge-cutting proteases of other bacteria . Protease production may be a virulence mechanism for P . multocida strains.

Infect Immun, 1992 Aug, 60(8), 3238 - 43
Pasteurella haemolytica leukotoxin enhances production of leukotriene B4 and 5-hydroxyeicosatetraenoic acid by bovine polymorphonuclear leukocytes; Henricks PA et al.; The influence of the leukotoxin of Pasteurella haemolytica on the generation of arachidonic acid metabolites by bovine polymorphonuclear leukocytes (PMNs) was investigated . PMNs released 5-, 12-, and 15-hydroxyeicosatetraenoic acids (5-, 12-, and 15-HETE) and leukotriene B4 (LTB4) upon stimulation with arachidonic acid . The leukotoxin preparations dose dependently enhanced the release of the 5-lipoxygenase products 5-HETE and LTB4 in arachidonic acid-stimulated PMNs, whereas the release of 12- and 15-HETE was not affected . The enhanced release of LTB4 and 5-HETE was not due to a decreased cellular retention of the 5-lipoxygenase products . In addition, leukotoxin preparations by themselves were also able to induce LTB4 and 5-HETE production in the absence of exogenous arachidonic acid . Generation of 5-lipoxygenase products by PMNs stimulated by leukotoxin may represent an important cellular event that occurs during infections with P . haemolytica.

J Am Vet Med Assoc, 1992 Jul 15, 201(2), 326 - 8
Chronic frontal sinusitis in dairy cattle: 12 cases (1978-1989); Ward JL et al.; Chronic frontal sinusitis in 12 dairy cattle most often was associated with a history of dehorning, in which the sinus was entered (67%), or with respiratory tract disease (25%) . The most common organisms isolated were Actinomyces pyogenes and Pasteurella multocida . Signs of infection did not develop for months in some cattle and were often intermittent . The most common clinical signs included anorexia, lethargy, fever, frontal bone distortion, exophthalmos, abnormal posture, nasal discharge, and neurologic abnormalities . Treatment consisted of trephination at 2 sites, drainage and lavage of the sinus cavity, and administration of antibiotics and analgesics . Eight cattle responded well to treatment and were discharged, but 4 others had signs of CNS involvement and died or were euthanatized . Trephination of the frontal sinus cavity at carefully chosen sites and antibiotic treatment are indicated when sinusitis is suspected . Drainage of the sinus cavity is imperative to avoid extension of the infection into the CNS.

Infect Immun, 1992 Jul, 60(7), 2726 - 32
Molecular cloning and expression of ptxA, the gene encoding the 120-kilodalton cytotoxin of Actinobacillus pleuropneumoniae serotype 2; MacDonald J et al.; The genetic determinants of the 120-kDa cytotoxin of Actinobacillus pleuropneumoniae serotype 2 were isolated from a lambda DNA library by a plaque immunoblot technique . Expression of the 120-kDa polypeptide was confirmed by Western immunoblot analysis of infected Escherichia coli cell lysates, which were shown to be toxic for porcine alveolar macrophages in vitro . The genetic determinants of the toxin were subcloned into the plasmid vector pUC18 . This plasmid (pPTX1) directed the synthesis and secretion of the active 120-kDa cytotoxin in E . coli . The recombinant toxin was indistinguishable from native cytotoxin from A . pleuropneumoniae serotype 2 with respect to molecular size, reaction in Western blot analysis, heat lability, cytotoxic activity, and neutralization by serum antibody . A restriction endonuclease cleavage map of pPTX1 was prepared, and deletion mutants were used to locate the minimal region of DNA required for production of intracellular toxin; this gene was termed ptxA . Southern hybridization analysis with a 1.7-kb PvuII fragment located within the ptxA gene revealed sequences with a high degree of homology in serotype reference strains 2, 3, 4, 6, and 8 . Other reference strains did not contain sequences that were recognized by this probe . However, related sequences (greater than 71% homology) were detected in Actinobacillus actinomycetemcomitans and A . equuli . Weak hybridization was observed between the ptxA probe and pLKT5, which carries the lktAC genes of Pasteurella haemolytica, and between the ptxA probe and pAPH1, which carries the structural gene for type II hemolysin from A . pleuropneumoniae . The isolation of the genetic determinants of this cytotoxin will enable investigations of the structure and organization of the ptx DNA region and further analysis of its role in the pathogenesis of pleuropneumonia.

J Wildl Dis, 1992 Jul, 28(3), 435 - 42
Serologic survey for selected arboviruses and other potential pathogens in wildlife from Mexico; Aguirre AA et al.; During 1988 and 1989, a serologic survey of wildlife was conducted in northeastern Mexico to determine the presence, prevalence, and distribution of arboviruses and other selected disease agents . Eighty mammal specimens were tested . Antibodies to vesicular stomatitis-Indiana, Venezuelan equine encephalitis-Mena II, Rio Grande virus, and vesicular stomatitis-New Jersey were detected predominantly in small mammals . Deer and mouflon (Ovis musimon) had antibodies to bluetongue and epizootic hemorrhagic disease . Two species had serologic evidence of recent exposure to Francisella tularensis . A white-tailed deer (Odocoileus virginianus) had antibodies to Anaplasma marginale . All specimens tested for antibodies against Yersinia pestis and Brucella abortus were negative . Sera from 315 birds were tested for antibody against five equine encephalitis viruses and six avian pathogens . During 1988, antibodies to Venezuelan equine encephalitis-Mena II, Venezuelan equine encephalitis-TC83, St . Louis encephalitis, eastern equine encephalitis, and western equine encephalitis were detected in birds of several species . Antibodies to Pasteurella multocida and Newcastle disease virus were also detected . Birds from five species presented antibodies to Mycoplasma meleagridis . Specimens tested for M . gallisepticum, M . synoviae, and Chlamydia psittaci were negative . To the best of our knowledge, this survey represents the first serologic evidence of bluetongue, Cache Valley virus, epizootic hemorrhagic disease, Jamestown Canyon virus, vesicular stomatitis-Indiana, vesicular stomatitis-New Jersey, Rio Grande virus, and tularemia reported among wildlife in Mexico.

Berl Munch Tierarztl Wochenschr, 1992 Jul 1, 105(7), 233 - 5
{The importance of dermonecrotoxin and dermonecrotoxoid of Pasteurella multocida for the calf}; Schimmel D et al.; The i . m . application of dermonecrotoxin of P . multocida var . D led to an atrophy of nasal conchae, liver-swelling and to induration of liver and spleen . The intratracheal application caused a lobular pneumonia . The vaccination with toxoid resulted in an immunity against a challenge with toxin but not with P . multocida var . D . Correlations between antitoxic serum-titers and immunity have to be investigated in future.

Am J Vet Res, 1992 Jul, 53(7), 1108 - 12
Polypeptides associated with Pasteurella multocida infection in rabbits; Zimmerman TE et al.; Polypeptides from whole cell preparations of Pasteurella multocida serotypes A:12 and A:3 were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and transferred to nitrocellulose paper . Antigens were detected by immunoblot analysis, using sera from 3 groups of rabbits . Sera were obtained from rabbits inoculated intranasally with P multocida serotype A:12 or A:3, from rabbits maintained in a rabbitry with enzootic P multocida A:12 infection, and from rabbits maintained in a rabbitry with enzootic P multocida A:3 infection . Immunoblot analyses of pre- and postinoculation sera from experimentally infected rabbits, using serotype A:12 antigen, revealed 3 polypeptides with approximate molecular mass of 28, 30, and 37 kDa that consistently detected antibodies after P multocida-induced infection . Sera from rabbits naturally infected with either serotype, tested against serotype A:12 and A:3 antigens, detected the same polypeptides in both serotypes . Thus, immunologic reactivity to these polypeptides may be useful for serologic detection of P multocida infection.

Can J Vet Res, 1992 Jul, 56(3), 260 - 4
Enhanced adherence of Pasteurella multocida to porcine tracheal rings preinfected with Bordetella bronchiseptica; Dugal F et al.; Adherence of 25 isolates of Pasteurella multocida to porcine tracheal rings was evaluated . Results indicated that adherence was not related to the isolate's origin, capsular or somatic types, dermonecrotoxin production or hemagglutination activity . The effect of a preinfection with Bordetella bronchiseptica on the colonization by P . multocida was then studied . On rings infected with P . multocida alone, bacteria initially adhered to the epithelium, but within a few hours, the level of colonization decreased progressively . On rings preinfected with B . bronchiseptica, or pretreated with a cell-free B . bronchiseptica culture supernate (or filtrate), a high level of P . multocida colonization was maintained for at least 24 hours . Results indicate that B . bronchiseptica appears to facilitate upper respiratory tract colonization by P . multocida by a process which involves a low molecular weight (less than or equal to 1000) heat-stable substance, possibly the tracheal cytotoxin.

Can J Vet Res, 1992 Jul, 56(3), 199 - 203
Effects of Pasteurella haemolytica leukotoxic culture supernatant on bovine neutrophil aggregation; Conlon P et al.; Pasteurella haemolytica A1 leukotoxic culture supernatant was evaluated for its ability to cause aggregation of bovine peripheral neutrophils . Neutrophils were isolated by a hypotonic lysis method and incubated with zymosan-activated plasma (ZAP), leukotoxic culture supernatant, antileukotoxin serum, calcium and magnesium-free media, p-bromophenacyl bromide and protein kinase C inhibitors . Aggregation was evaluated by changes in infrared light transmittance . Leukotoxic culture supernatant caused neutrophils to aggregate, and this effect was significantly removed by preincubation with antileukotoxin serum . Aggregation to ZAP and leukotoxin was dependent on the presence of extra-cellular calcium . Activation of protein kinase C by phorbol myristate acetate induced aggregation which was reduced by staurosporine; however, aggregation to leukotoxin did not involve protein kinase C activation . Phospholipase A2 inhibition did not alter the aggregation response to ZAP or to leukotoxin . The in vitro measurement of neutrophil aggregation induced by the leukotoxin of P . haemolytica reflects cytoskeletal and other activation events that may contribute to the intense inflammatory process which this organism induces in the lungs of cattle.

Avian Dis, 1992 Jul-Sep, 36(3), 803 - 7
Severe mortality in broiler chickens associated with Mycoplasma synoviae and Pasteurella gallinarum; Droual R et al.; Severe economic loss due to high mortality and condemnation rates occurred on two commercial broiler facilities . Chickens had moderate-to-severe airsacculitis, pericarditis, perihepatitis, tracheitis, and synovitis . Pasteurella gallinarum was isolated from 16 of 18 pericardia, four of 14 livers, 11 of 16 air sacs, six of seven joints and one of 28 tracheas in pure culture . In addition, Mycoplasma synoviae was isolated from trachea and air sac . Lesions were suggestive of an Escherichia coli septicemia, but E . coli was isolated from only four of 28 tracheas and one of 14 livers in pure culture . A coronavirus was isolated from trachea and lung . Whether this coronavirus represented a vaccine or field strain of infectious bronchitis was not determined . These findings suggested that the severe lesions were due to a concomitant infection with an atypical strain of P . gallinarum.

Avian Dis, 1992 Jul-Sep, 36(3), 693 - 9
An atypical strain of Pasteurella gallinarum: pathogenic, phenotypic, and genotypic characteristics; Droual R et al.; The pathogenicity of a strain of Pasteurella gallinarum isolated in Fresno County, Calif., was compared with the American Type Culture Collection (ATCC) strain . Broiler chickens were inoculated intranasally with 10(7) colony-forming units (CFU) and intramuscularly with 10(5) CFU of each strain . The only notable lesions were in chickens inoculated intramuscularly with 10(5) CFU of the Fresno strain, which developed severe myositis at the inoculation site, pericarditis, perihepatitis, airsacculitis, and synovitis . P . gallinarum was reisolated from these lesions . Phenotypic characteristics of the two strains were identical except in reactions in ONPG broth and fermentation of xylose . Protein-banding patterns for the two strains were identical except for a single band difference in the 35-kilodalton region . Restriction endonuclease analysis confirmed that the Fresno strain was a distinct one . Plasmid analysis revealed that the ATCC strain had two plasmids and the Fresno strain had none.

Jikken Dobutsu, 1992 Jul, 41(3), 379 - 81
{A trial designed to obtain a specific pathogen free Syrian hamster colony by administration of chemicals}; Shibuya M et al.; Conventional Syrian hamsters, contaminated with Giardia spp., Spironucleus muris, Trichomonas spp., Pasteurella pneumotropica and Pseudomonas aeruginosa were treated with chemicals in order to obtain specific pathogen free animals . Hamsters kept in the laminar flow rack were treated orally with metronidazole several times to obtain a flagellate-free colony . After all flagellates had been eradicated, one pair of animals were kept in an isolator and mating was allowed to occur . When their offspring reached the age of seven weeks, they were intramuscularly injected daily with netilmicin sulfate for 10 consecutive days . Following these treatments, all of the hamsters were free of Pasteurella and Pseudomonas . Further breeding of these animals was continued in isolators . To confirm the absence of selected pathogens, they were placed in a barrier room for further breeding as specific pathogen free animals.

Acta Obstet Gynecol Scand, 1992 Jul, 71(5), 384 - 7
Pasteurella multocida chorioamnionitis from vaginal transmission; Wong GP et al.; A 21 year old primigravida with a twin pregnancy developed Pasteurella multocida chorioamnionitis . Infection occurred at 27 weeks gestational age after prolonged rupture of membranes . The twin in the separate sac presenting proximal to the cervix suffered infection and died shortly after birth whereas the other twin was not infected . The bacterium is believed to have caused ascending infection from asymptomatic colonization of the vaginal tract.

J Wildl Dis, 1992 Jul, 28(3), 347 - 54
Using ribosomal RNA gene restriction patterns in distinguishing isolates of Pasteurella haemolytica from bighorn sheep (Ovis canadensis); Snipes KP et al.; Pasteurella haemolytica isolates (n = 31) from two isolated captive herds of Rocky Mountain bighorn sheep (Ovis canadensis canadensis) were characterized and compared phenotypically (biotype, serotype, hemolytic activity) and by a genomic fingerprinting method known as ribotyping . Seven to nine distinct phenotypes were observed . Depending on the method used for serotyping, one to three phenotypes were common to both herds . Eighteen isolates, recovered from both herds, were non-hemolytic, biotype T, indirect hemagglutination assay serotype 4 . Ribotyping, a method for highlighting genetically conserved deoxyribonucleic acid restriction site heterogeneity with a 32P-labelled Escherichia coli ribosomal ribonucleic acid probe, produced six to eight distinct ribotype pattern groups within the 31 P . haemolytica isolates, depending on the restriction enzyme used . In contrast to phenotypes, ribotypes appeared unique to each herd, and ribotyping helped to further differentiate some isolates of the same biotype and serotype . In addition, ribotyping provided an alternative means for evaluating relationships between isolates differing in hemolytic activity but which were otherwise phenotypically identical . We propose that ribotyping may be a useful adjunct to other bacterial characterization methods in studying the epizootiology of pasteurellosis in bighorn sheep.

Vet Rec, 1992 Jun 20, 130(25), 549 - 53
Evaluation of an atrophic rhinitis vaccine under controlled conditions; Voets MT et al.; A vaccine containing inactivated cultures of Bordetella bronchiseptica, toxigenic Pasteurella multocida type D and dermonecrotic P multocida type D toxoid in an oil-in-water adjuvant was given to seven sows, with seven others acting as controls . Half the piglets in each litter were exposed intranasally when four days old to B bronchiseptica and when eight days old to toxigenic P multocida type D . There was considerably less sneezing in the litters of the vaccinated sows and when the piglets were 10 weeks old, only 18 per cent had deformed snouts compared with 74 per cent in the litters of the control sows . The average liveweight gain of the piglets born to vaccinated sows was significantly better (P less than 0.05) between two and 10 weeks of age than that of the piglets born to unvaccinated sows, although there were no significant lower respiratory tract lesions in either group . The conchal atrophy scores were significantly lower (P less than 0.001) in the piglets from the vaccinated sows and were negatively correlated (r = -0.37) with increasing liveweight gain . In the liters of the vaccinated sows, P multocida was not isolated from the nasal passages of the in-contact piglets and from only 7 per cent of those deliberately exposed compared with 65 per cent and 79 per cent, respectively, in the litters of the control sows . P multocida was isolated post mortem from the tonsils of 23 per cent of the piglets of vaccinated sows and from 87 per cent of those from unvaccinated sows.

Vet Microbiol, 1992 Jun 15, 31(4), 369 - 78
Factors affecting endotoxin release from the cell surface of avian strains of Pasteurella multocida; Lee MD et al.; Two avian strains of Pasteurella multocida, a vaccine strain and a virulent field isolate, were investigated to determine their propensity to release endotoxin from the cell surface . Both organisms released comparable amounts of endotoxin when plasma complement proteins were present, however the virulent strain did so without the loss of viability that occurred in the vaccine strain . Blocking complement activity decreased the ability of plasma to elicit endotoxin release from the bacteria . When the cells were treated with divalent metal chelators such as trans-1, 2-diaminocyclohexane-N,N,N1,N1-tetraacetic acid (CDTA), more endotoxin was released from the vaccine strain than from the virulent isolate . Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) of purified lipopolysaccharide (LPS) from both strains revealed virtually identical patterns . Both had patterns considered typical of rough LPS . Challenge studies in 8 weeks old turkeys showed that the field strain induced endotoxemia of longer duration than the vaccine strain and produced greater mortality.

Vet Immunol Immunopathol, 1992 Jun, 33(1-2), 155 - 62
Use of rats to compare atrophic rhinitis vaccines for protection against effects of heat-labile protein toxin produced by Pasteurella multocida serogroup D; Thurston JR et al.; Four bacterin-toxoid and three bacterin commercial vaccines against atrophic rhinitis were tested in rats for their capacity to immunize against the lethal and systemic effects of purified heat-labile protein toxin (D-toxin) produced by Pasteurella multocida serogroup D . Only one bacterin-toxoid vaccine stimulated sufficient immunity to prevent the death of all rats challenged with D-toxin . None of the vaccines prevented weight loss, leukocytosis or increases in serum complement titers in rats challenged with D-toxin . Rats provide an inexpensive animal model for testing the capacity of vaccines to generate antitoxic immunity against the lethal and systemic effects of D-toxin.

Vet Microbiol, 1992 Jun 1, 31(2-3), 161 - 8
Characterisation and biological activity of monoclonal antibodies specific for Pasteurella haemolytica A1 capsule and lipopolysaccharide; Wilson CF et al.; Monoclonal antibodies (mAb) against both Pasteurella haemolytica A1 capsule and lipopolysaccharide (LPS) were produced . Anti-capsule mAb reacted with the homologous A1 serotype only, whereas mAb against LPS reacted with P . haemolytica serotypes A2, A5, A8, A12, A14 and A16 but not with 33 bacterial species or rough LPS mutant strains tested . Both capsule and LPS antigens were visualised on the surface of bacteria by immunogold electron microscopy . Neither of the mAbs demonstrated antibody-dependent complement-mediated killing in vitro but both facilitated phagocytosis in vitro.

J Clin Microbiol, 1992 Jun, 30(6), 1398 - 401
Characterization of Pasteurella multocida from nasal cavities of piglets from farms with or without atrophic rhinitis; Lariviere S et al.; A total of 137 strains of Pasteurella multocida isolated from the nasal tracts of pigs with and without clinical atrophic rhinitis (AR) were studied for their biochemical, antigenic, and toxigenic characteristics . There were no major biochemical differences among the P . multocida isolates . Capsular antigen types A and D were both present in the nasal cavities of the pigs with or without clinical AR . However, the prevalence of type D was higher on farms with pigs with AR . Types A and D with different somatic antigens could both be present in the same pig . There was no correlation between somatic types and/or capsular types with the clinical AR status of the pigs on the farm . Toxigenic isolates were found only in pigs which had a problem of clinical AR, and a great majority of these isolates belonged to type D . Since there was a high level of heterogeneity of the strains in the P . multocida population on a farm, several strains should be characterized before the diagnosis of AR could be excluded on the basis of the absence of isolation of rhinopathogenic P . multocida strains.

J Arthroplasty, 1992 Jun, 7(2), 157 - 60
Pasteurella multocida infection in total knee arthroplasty . Case report and literature review; Guion TL et al.; Pasteurella multocida, a small gram-negative bacterium, is part of the normal mouth flora of many animals, including domestic cats and dogs . While commonly associated with infections in animals, it is a rare cause of human disease . The majority of Pasteurella infections in humans occur with percutaneous inoculation of the organism following a bite by a cat or dog, although disease without antecedent animal exposure or with causal animal contact does occur . The spectrum of disease produced ranges from localized, including abscess, cellulitis, lymphadenopathy, and osteomyelitis, to systemic, with septicemia, septic arthritis, respiratory, and central nervous system involvement . Altered host defenses and underlying chronic disease, such as rheumatoid arthritis, corticosteroid therapy, and severe hepatic or renal disease, may predispose to more serious systemic manifestations of infection . The authors report a case of P . multocida infection in a total knee arthroplasty as a result of a dog scratch and review the literature reporting P . multocida infections in total knee arthroplasty.

J Leukoc Biol, 1992 Jun, 51(6), 579 - 85
Tumor necrosis factor alpha and interleukin 1 alpha enhance lipopolysaccharide-mediated bovine endothelial cell injury; Sharma SA et al.; Alveolar macrophages (AMs) are important in the host response to aerogenous pulmonary bacterial infections, such as Pasteurella haemolytica-induced pneumonia in cattle . Previous work has shown that AMs enhance P . haemolytica-mediated pulmonary endothelial cell (EC) damage in vitro . The purpose of this study was to determine the mechanism of AM-enhanced EC damage using an in vitro AM-EC coculture system consisting of AMs cultured on culture plate insert membranes and ECs in the underlying chamber . The addition of lipopolysaccharide (LPS) to the culture plate insert chamber resulted in EC damage indicated by 51Cr release, which was enhanced in the presence of AMs . To determine the role of AM-secreted cytokines, recombinant human interleukin 1 alpha (IL-1) or tumor necrosis factor alpha (TNF) was added to ECs simultaneously with varying concentrations of LPS . Although TNF and IL-1 alone had only marginal toxic effects on ECs, the simultaneous treatment of TNF or IL-1 with LPS greatly increased the LPS cytotoxic effect on ECs . In addition, IL-1 receptor antagonist eliminated the IL-1 enhancement of LPS-mediated EC toxicity . These results suggest that macrophage-secreted cytokines synergistically enhance LPS-mediated pulmonary EC damage.

Infect Immun, 1992 Jun, 60(6), 2166 - 73
Molecular characterization of an RTX toxin determinant from Actinobacillus suis; Burrows LL et al.; RTX cytolysins are a family of calcium-dependent, pore-forming, secreted toxins found in a variety of gram-negative bacteria . The prototypical member of this family is the alpha-hemolysin of Escherichia coli . The RTX genetic determinants from seven members of the family Pasteurellaceae, Pasteurella haemolytica, Actinobacillus actinomycetemcomitans, and A . pleuropneumoniae serotypes 1,5,7, and 9 were previously cloned and sequenced . Using the leukotoxin determinant from P . haemolytica serotype A1 as a probe, we detected the presence of RTX-type determinants in Actinobacillus suis, A . equuli, and A . lignieresii of the family Pasteurellaceae . All three species elaborate proteins of approximately 104 to 110 kDa that are recognized by polyclonal antisera against the 104-kDa hemolysin of A . pleuropneumoniae serotype 1 . An RTX determinant of A . suis isolate 3714 was cloned and sequenced and was found to be almost identical to the RTX determinant of A . pleuropneumoniae serotypes 5 and 9 . In addition, the determinant is not composed of four contiguous genes, as had been reported for most other RTX determinants; instead, the genes encoding the two proteins responsible for secretion of the toxin are at a locus distinct from that containing the toxin structural and activation genes.

Microb Pathog, 1992 Jun, 12(6), 459 - 63
Pasteurella haemolytica leukotoxin inhibits mitogen-induced bovine peripheral blood mononuclear cell proliferation in vitro; Czuprynski CJ et al.; In this study we demonstrate that partially purified Pasteurella haemolytica leukotoxin inhibits the proliferative response of bovine peripheral blood mononuclear cells (PBMC) to mitogens in vitro . Inhibition of PBMC proliferation did not appear to be due to cell death . Addition of a neutralizing anti-leukotoxin monoclonal antibody restored a normal proliferative response.

J Vet Med Sci, 1992 Jun, 54(3), 403 - 7
Toxigenic type A Pasteurella multocida as a causative agent of nasal turbinate atrophy in swine; Sakano T et al.; Although no clinical signs of atrophic rhinitis (AR) were recognized in 2- and 5-week-old pigs, approximately 60% of 2- to 6-month-old pigs showed clinical signs of AR in an affected pig farm . None of the pigs had normal turbinate at slaughter . Bordetella bronchiseptica was not isolated from any of the pigs before onset and incipient stage of the outbreak (2-week to 2-month-old) . Pasteurella multocida of capsular type D was not isolated from any of those pigs . However, toxigenic P . multocida of capsular type A was isolated from a number of the pigs immediately before onset and incipient stage of the outbreak . Thirty-six-day-old primary specific-pathogen-free pigs were inoculated intranasally with a toxigenic type A P . multocida isolated from a 5-week-old pig . Severe nasal turbinate atrophy was observed in those pigs which were necropsied at 3 weeks post-inoculation . This is the first report on outbreak of severe nasal turbinate atrophy induced by toxigenic type A P . multocida in Japan.

Vet Microbiol, 1992 Jun 1, 31(2-3), 197 - 206
Experimental atrophic rhinitis in 2 and 4 month old pigs infected sequentially with Bordetella bronchiseptica and toxigenic type D Pasteurella multocida; Sakano T et al.; Experimental infections with Bordetella bronchiseptica and/or toxigenic type D Pasteurella multocida were studied in 2- and 4-month-old primary specific-pathogen-free pigs . None of the 2-month-old pigs inoculated with B . bronchiseptica or P . multocida alone developed turbinate atrophy . All the pigs inoculated with B . bronchiseptica (10(7) CFU/head) and P . multocida (10(9) CFU/head for 5 consecutive days) together, however, developed clinical and post-mortem signs of atrophic rhinitis (AR) similar to the naturally occurring disease . Slight to severe turbinate atrophy was observed in the 4-month-old pigs inoculated with B . bronchiseptica and P . multocida (at the same concentration as above) at necropsy.

Am J Vet Res, 1992 Jun, 53(6), 971 - 5
Comparison of antibody responses in cattle to outer membrane proteins from Pasteurella haemolytica serotype 1 and from eight untypeable strains; Simons KR et al.; Membrane associated proteins from 8 untypeable Pasteurella haemolytica strains were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and compared with those of P haemolytica serotypes 1 and 2 . Cattle antisera obtained from P haemolytica serotype 1 vaccine trials were used in immunoblotting assays to compare the membrane proteins from the 8 untypeable strains with those from P haemolytica serotypes 1 and 2 . Densitometry was used to identify bands, and using linear regression analyses, the peak area optical densities (measuring antibody response) were correlated to lesion scores from the vaccinated calves . Significant antibody responses to proteins of 99, 69, 60, 55, 47, 45, 39, 33, 30, 16, and 14.5 kDa were detected for 4 or more of the 8 P haemolytica untypeable strains . Serotypes 1 and 2 of P haemolytica contained a comigrating 30-kDa protein . Antibody responses to proteins of 39, 33, and 32.5 kDa were significant for 3 of the untypeable strains and had significant correlation to lesion scores . Antibody responses to various other proteins were significant for 2 untypeable strains each.

J Clin Microbiol, 1992 Jun, 30(6), 1518 - 24
Comparison of DNA fingerprints and somatic serotypes of serogroup B and E Pasteurella multocida isolates; Wilson MA et al.; The DNA fingerprint profiles and somatic serotypes of 71 Pasteurella multocida capsule serogroup B isolates, 13 capsule serogroup E isolates, and 16 somatic reference serotype strains were compared . Each of the 16 reference somatic serotypes had a unique DNA fingerprint profile with the HhaI restriction endonuclease . Fifty-four serogroup B isolates (isolated from classical cases of hemorrhagic septicemia) reacted with somatic serotype 2 or 5 antiserum and had DNA fingerprint profiles which resembled that of the serotype 2 reference strain . Seven DNA fingerprint profiles were found among 16 serogroup B strains representing other somatic serotypes . The DNA fingerprints of these isolates were different from the fingerprints of the 16 somatic reference serotype strains . All 13 serogroup E isolates had identical somatic serotypes and identical DNA fingerprint profiles when the HhaI endonuclease was used . The HhaI fingerprint profile of the serogroup E isolates did not match any fingerprint profile of the reference somatic serotype strains . Following DNA profiling with the HhaI endonuclease, the 13 serogroup E isolates were differentiated sequentially with HpaII restriction endonuclease . A descriptive identification epithet for P . multocida isolates was constructed . The descriptive epithet consists of serologic identification and sequential DNA profiles with restriction endonucleases HhaI and HpaII, respectively . DNA fingerprinting of P . multocida is a precise characterization method . In conjunction with serologic typing, it can further classify P . multocida isolates for epidemiologic studies.

Exp Hematol, 1992 Jun, 20(5), 590 - 9
Differential sensitivity of CD34 epitopes to cleavage by Pasteurella haemolytica glycoprotease: implications for purification of CD34-positive progenitor cells; Sutherland DR et al.; Our previous studies have shown that a unique glycoprotease from Pasteurella haemolytica specifically cleaves only proteins containing sialylated O-linked glycans . The hematopoietic progenitor cell antigen, CD34, which is heavily glycosylated with both N- and O-linked glycans, is readily cleaved by this protease . In this study, we demonstrate that the epitopes detected by five of the seven CD34 monoclonal antibodies are removed by the glycoprotease . The differential sensitivity of the CD34 epitopes to cleavage with either neuraminidase and/or glycoprotease establishes three classes of epitopes: 1) (class I) those identified by MY10, B1.3C5, 12.8, and ICH3 that are differentially affected by neuraminidase and removed by the glycoprotease; 2) (class II) the epitope detected by QBEND 10 that is removed only by the glycoprotease; and 3) (class III) those identified by TUK3 and 115.2 that are not removed by either enzyme . Cleavage of the 110-kd CD34 structure by the glycoprotease generates a major cell-bound fragment of about 75 kd, identified by the class III antibodies . We have also used the enzyme to improve the rapid recovery of CD34+ cells selected by immunomagnetic affinity techniques . In a preclinical model, we separated CD34+ KG1 cells with high yield (90%-95%) and high purity (94%-98%) from sham mixtures containing 50% CD34- cells . We also separated CD34+ blast cells from a patient in megakaryoblastic crisis of chronic myelogenous leukemia . In this case, the purity and yield were 93% and 94%, respectively . Enzyme treatment had no detrimental effect on cell viability, and the treated cells showed a normal quantitative expression and distribution of CD34 antigen as assessed with class III antibodies . We conclude that the P . haemolytica glycoprotease has potential to improve the isolation, from human bone marrow, of primitive hematopoietic cells that carry the CD34 antigen.

Vet Immunol Immunopathol, 1992 Jun, 33(1-2), 51 - 68
Effects of Pasteurella haemolytica A1 leukotoxin on bovine neutrophils: degranulation and generation of oxygen-derived free radicals; Maheswaran SK et al.; To further define the role of Pasteurella haemolytica A1 leukotoxin in the pathogenesis of bovine pneumonic pasteurellosis, its in vitro effects on bovine neutrophils were investigated . Leukotoxin-containing culture supernatant, from P . haemolytica, stimulated a neutrophil respiratory burst as measured by the generation of oxygen-derived free radicals O2- and H2O2 . This effect was immediate because preincubation of neutrophils with the culture supernatant for 5 min or longer substantially suppressed this respiratory burst . This suppression was due to cytolysis of the neutrophils . Prolonged incubation of neutrophils with the same culture supernatant caused further cytolysis and degranulation . Heat-inactivated P . haemolytica culture supernatant that had lost its cytotoxic properties failed to stimulate respiratory burst by neutrophils . Furthermore, the respiratory burst, cytolysis and degranulation were abrogated only by leukotoxin-neutralizing monoclonal and polyclonal antibodies, but not by antibodies against the lipopolysaccharide . These studies show that the leukotoxin component in the culture supernatant was responsible for the generation of oxygen-derived free radicals and proteolytic enzymes from neutrophils which may participate in direct lung injury.

Proc Natl Acad Sci U S A, 1992 May 15, 89(10), 4240 - 4
Pasteurella multocida toxin is a potent inducer of anchorage-independent cell growth; Higgins TE et al.; The growth of many normal cells requires contact with an adhesive substratum, a requirement that is frequently abrogated in the transformed phenotype . We have explored pathways that can lead to the anchorage-independent growth of cultured Rat-1 fibroblasts . Pasteurella multocida toxin (PMT), a 146-kDa mitogenic protein, caused a striking increase in the formation of colonies (greater than 200 microns) from single cells in soft agar . The magnitude of the effect of PMT was greater than that achieved by epidermal growth factor or platelet-derived growth factor . The toxin was extremely potent, with half-maximal and maximal effects observed at 1 and 10 pM PMT, respectively . This concentration dependence of the action of the toxin is similar to that for the stimulation of DNA synthesis in adherent cultures of the cells . Stimulation of colony formation could be achieved by a transient exposure of the cells to PMT and it was blocked by methylamine, indicating that the toxin enters the cells to act . Colony formation was stimulated equally by native and recombinant PMT, but a truncated version (33.5 kDa) of the recombinant toxin was ineffective . PMT antiserum blocked colony formation in response to PMT . In the Rat-1 cells, PMT stimulated the phospholipase C-mediated hydrolysis of inositolphospholipids, as indicated by the stimulation of inositol phosphate release, Ca2+ mobilization, and phosphorylation of a protein kinase C substrate . The results indicate that the deregulation of signal-transduction pathways as elicited by an intracellularly acting bacterial toxin can induce a malignant phenotype.

Infect Immun, 1992 May, 60(5), 1734 - 9
Characterization of a neutralizing monoclonal antibody to Pasteurella haemolytica leukotoxin; Gerbig DG Jr et al.; Six hybridoma clones producing monoclonal antibodies (MAbs) reactive with Pasteurella haemolytica A1 leukotoxin were derived from mice immunized with leukotoxin excised from sodium dodecyl sulfate-polyacrylamide gels . Of the six MAbs, only one, Ltx-2, neutralized leukotoxin in a BL-3 cell cytotoxicity assay . MAb Ltx-2 blocked association of A1 leukotoxin to BL-3 cells, as measured by flow cytometric analysis . The epitope recognized by Ltx-2 was localized to the carboxyl half of the native protein, between residues 450 and 939, by Western immunoblot analysis of CNBr fragments . Further analysis with leukotoxin deletion proteins indicated either that the Ltx-2-reactive epitope was localized in the carboxyl portion of the leukotoxin between amino acids 768 and 939 or that this region influences MAb recognition of the epitope . MAb Ltx-2 was tested for neutralizing activity against leukotoxin produced by P . haemolytica serotypes 1 through 12 . The MAb neutralized leukotoxin produced by all of the A biotype isolates (serotypes 1, 5, 6, 7, 8, 9, and 12), with the exception of serotype A2, but did not neutralize any T biotype leukotoxin tested (T3, T4, or T10) . The results indicate that MAb Ltx-2 neutralizes leukotoxin by interfering with target cell association and that the MAb-specific epitope is either not present or not critical for function in the leukotoxin produced by P . haemolytica serotypes A2, T3, T4, and T10.

J Gen Microbiol, 1992 May, 138 ( Pt 5), 909 - 22
Outer-membrane protein and lipopolysaccharide variation in Pasteurella haemolytica serotype A1 under different growth conditions; Davies RL et al.; Growth characteristics, as well as outer-membrane protein (OMP) and lipopolysaccharide (LPS) profiles in SDS-polyacrylamide gels, of two serotype A1 isolates of Pasteurella haemolytica were examined under different in vitro growth conditions . The two isolates were chosen as representatives of disease (S/C 82/1) and non-disease (W/D 83/4) isolates, respectively . The growth rates and final cell densities of both isolates increased as the degree of aeration increased . In particular, the final cell densities varied significantly according to the degree of aeration . Under anaerobic conditions, however, both the growth rate and final cell density were significantly reduced . There was reduced expression of a 40.5 kDa protein under anaerobic conditions in both isolates, whereas in S/C 82/1 expression of the 71, 77 and 100 kDa iron-regulated proteins increased as aeration decreased . There were also differences in low-molecular-mass components of LPS between cells grown anaerobically and those grown aerobically . Growth in the presence of 5% CO2 did not significantly alter the growth rate and had little, if any, affect on OMPs or LPS . Differences in the expression of certain proteins occurred as growth progressed from the exponential to the stationary phase . Growth in the presence of the iron chelators 2,2'-dipyridyl, ethylenediamine-dihydroxyphenylacetic acid (EDDA), desferrioxamine mesylate (desferal), ovotransferrin (conalbumin) and bovine transferrin was inhibited within a very narrow concentration range . In the presence of 2,2'-dipyridyl, EDDA or desferal, 71 and 100 kDa iron-regulated OMPs increased in both isolates whereas a 77 kDa protein increased in isolate S/C 82/1 only . In the presence of ovotransferrin or bovine transferrin there was, in both isolates, increased expression of the 71 kDa protein, a slight increase in expression of the 100 kDa protein but no expression of the 77 kDa protein; there was also increased production of the 40.5 kDa protein, and synthesis of two additional proteins of 23 and 26 kDa . Other differences occurred after growth in foetal and newborn calf sera . In foetal calf serum there was enhanced expression of the 71 but not of the 100 kDa protein . In newborn calf serum there was no enhanced expression of the 71, 77 or 100 kDa proteins, but expression of novel proteins of 97 and 98 kDa as well as a high-molecular-mass protein occurred . There was also slight quantitative differences in the LPS profiles of cells grown in foetal or newborn calf sera compared to those of cells grown in other media.(ABSTRACT TRUNCATED AT 400 WORDS)

Dtsch Tierarztl Wochenschr, 1992 May, 99(5), 213 - 6
{The effect of the lactation period on the cell content of sheep milk}; Baumgartner W et al.; Milk samples of 201 ewes were examined in 6 week intervals during a complete lactation period . Those samples were analyzed for the presence of pathogenic bacteria and the somatic cell count was determined . Besides, the California Mastitis Test (CMT) was performed and the udder was clinically examined . The cell counts were found to depend on the lactation period . During 6 weeks following parturition the cell count was 63,000 cells/ml . This number decreased towards the 24th week of lactation to 32,000 cells/ml . At the end of lactation this value increased again to 425,000 cells/ml . The median value of ewes with normal udder health was 56,000 cells/ml milk . For samples from which pathogenic bacteria were isolated this value was 159,000 cells/ml . The most frequent pathogens isolated from the milk samples were coagulase-negative cocci (59.6% of bacteriologically positive samples), the median number being 88,000 somatic cells/ml in these sheep . Coagulase-positive cocci were isolated in 25.3% of the samples, the median value of the cell count was 295,000 cells/ml . In 12.1% of the samples streptococci were found . The median value was 167,000 cells/ml . From the remaining 3.0% of bacteriologically positive samples Pasteurellae, E . coli and Actinomycetae were isolated . The median value of the somatic cell count was 184,000 cells/ml . We consider coagulase-positive cocci therefore as the most pathogenic bacteria for the ovine udder.

Dtsch Tierarztl Wochenschr, 1992 May, 99(5), 204 - 6
{Results of experimental immunization of calves with different Pasteurella antigens}; Schimmel D et al.; Calves immunised with different Pasteurella antigens (inactivated whole cells, sodium chloride extract) where challenged two weeks after the second immunization with the homologous strain . The intracutaneous application of whole cells of P . haemolytica A1 and P . multocida A was effective . The incidence of pneumonia was reduced and the pneumonic lesions were less severe . The sodium chloride extract was not effective.

Aust Vet J, 1992 May, 69(5), 101 - 3
Vaccine efficacy for reducing turbinate atrophy and improving growth rate in piggeries with endemic atrophic rhinitis; Kabay MJ et al.; Two vaccines, based on formalin-killed whole cells of toxigenic Pasteurella multocida type D and Bordetella bronchiseptica combined with a partially toxoided cell extract of P multocida, were prepared with Freund's incomplete adjuvant (vaccine 1) or by alum precipitation (vaccine 2) . Each was tested for safety and efficacy in reducing the severity of nasal turbinate atrophy and improving the growth rate of pigs in three Western Australian commercial piggeries with endemic atrophic rhinitis . In safety experiments with vaccine 1, no adverse clinical effects were observed in vaccinated sows or their progeny . Piglets receiving vaccine 2 showed no injection site abnormalities, pyrexia or turbinate atrophy . In field trials, vaccine 1 significantly reduced the prevalence of moderate to severe nasal turbinate atrophy (Done score 3 to 5) when used in two piggeries (A and B) . Progeny from vaccinated sows in piggery B also grew significantly faster than controls . When vaccine 2 was used in piggery A at a later date and in another piggery (C), growth rate was not improved in either piggery and the prevalence of moderate to severe turbinate atrophy was reduced only in piggery C.

FEMS Microbiol Lett, 1992 May 1, 71(3), 211 - 6
Characterization of lipopolysaccharides from four Pasteurella haemolytica serotype strains: evidence for presence of sialic acid in serotypes 1 and 5; Utley SR et al.; Highly purified lipopolysaccharides (LPS) obtained from four strains of Pasteurella haemolytica representative of four different serotypes were studied to ascertain their overall structural elements and sugar and fatty acid compositions . SDS-PAGE analysis revealed that each LPS was of the smooth-type although they differed in migration patterns . Somewhat unusual features of these LPS included the presence of: (a) rhamnose in the core oligosaccharides of serotypes 2 and 3; and (b) sialic acid in the LPS of serotypes 1 and 5 . The fatty acids, myristic, hydroxymyristic and palmitic occur in essentially equivalent amounts in each of these LPS . In addition, stearic acid was present in small amounts of serotypes 1 and 5.

Orthop Rev, 1992 May, 21(5), 601, 604 - 5
Pasteurella infection in a total knee arthroplasty; Gabuzda GM et al.; Hematogenous infection of a total joint arthroplasty is a serious complication that has well-known etiologies . One of the most unusual inciting events is a bite wound . Pasteurella multocida is an anaerobic organism found in the mouths of mammals that has rarely been found to infect total knee arthroplasties . Prompt recognition of such an infection and prophylactic treatment with a penicillinase-resistant penicillin should maximize the patient's opportunity to eradicate such an infection.

Am J Vet Res, 1992 May, 53(5), 684 - 8
Response of Pasteurella haemolytica to erythromycin and dexamethasone in calves with established infection; Clarke CR et al.; A subcutaneous soft tissue infection model in calves was used to study the in vivo response of Pasteurella haemolytica to erythromycin and dexamethasone . Two tissue chambers were implanted SC in each of 12 calves . At 45 days after implantation, all tissue chambers were inoculated with an erythromycin-sensitive strain of P haemolytica . Starting 24 hours after inoculation, calves were allotted to 4 groups of equal size and a 2 x 2-factorial arrangement of treatments was applied: 3 calves were given erythromycin (30 mg/kg of body weight, IM, for 5 days), 3 calves were given dexamethasone (0.05 mg/kg, IM, for 2 days), 3 calves were given erythromycin and dexamethasone, and the remaining calves served as nontreated controls . Chamber fluids were tested daily, and the response to treatment was measured . Neither erythromycin nor dexamethasone affected viability or growth of bacteria within tissue chambers . Dexamethasone had no effect on the influx of neutrophils into infected chambers . Despite repeated administration of a high dose of erythromycin and attainment of adequate concentration in serum, erythromycin concentration in chamber fluids did not exceed the minimal inhibitory concentration established in vitro . These results indicate that the clinical efficacy of erythromycin against P haemolytica sequestered in consolidated pneumonic lesions may not be well correlated with predictions based on serum pharmacokinetic and in vitro susceptibility data.

Am J Vet Res, 1992 May, 53(5), 679 - 83
Use of an indwelling bronchial catheter model of bovine pneumonic pasteurellosis for evaluation of therapeutic efficacy of various compounds; Paulsen DB et al.; A model of bovine pneumonic pasteurellosis, using an indwelling bronchial catheter for inoculation and subsequent lavage of a single main stem bronchus of the lung, was evaluated in a preliminary efficacy trial of an experimental therapeutic compound . Inoculation of 10(7) Pasteurella haemolytica organisms into the bronchus consistently induced a focal pneumonic lesion with typical morphology of pneumonic pasteurellosis in the left or right caudal lung lobe . The experimental treatment caused significant (P less than 0.05) reduction in lung lesion volume, compared with that of a saline-treated control . It also caused significant (P less than 0.05) reduction in lavage fluid bacterial counts at 48 hours after inoculation, compared with counts in the controls . The inflammatory cell count and the percentage of neutrophils increased markedly in lavage fluids 8 hours after inoculation, but differences were not detected between treatments . Significant differences between treatments were not found in clinical signs, rectal temperature, or histologic changes . This model appears to be a sensitive indicator of treatment efficacy and has the advantage over previous models of pneumonic pasteurellosis of allowing sequential monitoring of the primary lesion site.

Am J Vet Res, 1992 May, 53(5), 670 - 3
Long-term study of aerobic bacteria of the genital tract in stud dogs; Bjurstrom L et al.; The aerobic bacterial flora of the genital tract was characterized in 15 stud dogs in an 18-month study . The dogs represented 4 breeds and were from 3 kennels . Bacterial samples from the prepuce and semen were collected every month, except in connection with matings, when they were collected weekly (464 samples) . The dogs that were included all mated at least once during the study . The mean pregnancy rate, litter size, and pup mortality for the bitches with which they had mated were all within normal limits . The most frequent bacteria isolated from the prepuce and semen were Pasteurella multocida, beta-hemolytic streptococci, and Escherichia coli . There was a tendency for breeds to differ in frequency of the most common bacterial species . Bacterial culture yielded no aerobic growth in 14.2% of the preputial samples and 69.8% of the semen samples . Bacteria were transferred between dog and bitch at mating . In this study of healthy breeding dogs, neither the fertility of the dog nor that of the bitch was affected by the bacteria transferred.

Am J Vet Res, 1992 May, 53(5), 665 - 9
Long-term study of aerobic bacteria of the genital tract in breeding bitches; Bjurstrom L et al.; The aerobic bacterial flora of the genital tract was characterized in 59 bitches in an 18-month study . The bitches represented 4 breeds and were from 3 kennels . Collection of vaginal swab specimens for bacterial culturing was performed every month, except during estrus when specimens were collected every week (n = 826) . The capsule of the swab containing transport media was broken before specimen collection to moisten the tip, which helped to reduce the number of negative cultures . All bitches helped at least once during the study and, thus, had known reproductive functions . Pregnancy rates, litter sizes, and pup mortality were within normal limits . Pasteurella multocida, beta-hemolytic streptococci group G, and Escherichia coli were the most common bacteria isolated . Although these species generally were isolated from mixed cultures, pure cultures were obtained from 18% of the specimens . There was a tendency for the various breeds to differ in their vaginal bacterial flora . The flora also varied during the reproductive cycle . Pasteurella multocida was isolated significantly more often during proestrus, estrus, metestrus, and pregnancy, than during anestrus and the postpartum period, and beta-hemolytic streptococci were isolated significantly more often during proestrus than during estrus, pregnancy, or the postpartum period . Staphylococcus intermedius was almost exclusively found after parturition . Culture results were negative for only 5.2% of specimens cultured . On the basis of our findings, bacterial culturing of vaginal swab specimens from bitches without signs of genital disease is of little value.

Am J Vet Res, 1992 May, 53(5), 646 - 52
Sequential development of antigens and toxins of Pasteurella haemolytica serotype 1 grown in cell culture medium; Confer AW et al.; Pasteurella haemolytica was grown in nonsupplemented cell culture medium, or in medium supplemented with bovine serum albumin (BSA) for 24 hours . The production of leukotoxin (LKT) and endotoxin was sequentially evaluated, as were bacterial antigens associated with bacterial cell lysates and culture supernates . Supplementation of medium with BSA had no effect on bacterial growth curves; however, LKT activity was detected earlier and was greater in culture supernates from BSA-supplemented media than from nonsupplemented medium . Leukotoxin antigen (105 kDa) was detected in culture supernates, using a monoclonal antibody, immunoblot analysis, and densitometry . The relative concentrations of LKT antigen were proportional to LKT activity . Endotoxin activity was initially lowest in the culture supernates from nonsupplemented medium, but increased during the incubation period, whereas endotoxin activity in BSA-supplemented culture supernates decreased with time in culture . In culture supernates from nonsupplemented medium, the number of antigenic bands identified by immunoblot analysis with hyperimmune anti-P haemolytica and densitometry was greater than in culture supernates from supplemented media . In bacterial lysates, a 95-kDa antigen was the major antigen detected, using the anti-LKT monoclonal antibody . The concentration of that antigen varied among lysates from nonsupplemented medium and BSA-supplemented media . Using hyperimmune anti-P haemolytica serum, minor differences were seen in the relative quantities of lysate-associated antigens dependent on time in culture and medium used . Among the major antigens seen, differences were most apparent for 150-, 100-, and 87-kDa antigens, whereas differences were not obvious for 42- 40-, and 30-kDa antigens.(ABSTRACT TRUNCATED AT 250 WORDS)

Am J Vet Res, 1992 May, 53(5), 631 - 5
Increased elastase activity in nasal mucus associated with nasal colonization by Pasteurella haemolytica in infectious bovine rhinotracheitis virus-infected calves; Briggs RE et al.; Four healthy calves were inoculated with Pasteurella haemolytica serotype 1 by instillation of a broth culture into the middle nasal meatus of the left nostril . Four weeks later, calves were exposed to infectious bovine rhinotracheitis virus by aerosol into both nostrils . All calves became ill, from approximately day 3 through day 10 after virus exposure, and shed increased amounts of nasal mucus . Two calves were induced to shed P haemolytica by the virus infection, and 2 calves required reinoculation with P haemolytica for nasal passages to become actively colonized . Elastase activity in nasal mucus increased about 15-fold within 3 days and peaked about 60-fold over baseline by 7 days after virus exposure . Activity of N-acetyl-beta-D-glucosaminidase, a measure of cell damage and serum leakage, increased slightly by day 3 and reached plateau on day 5, almost threefold over baseline activity . Protein and carbohydrate content increased at a rate similar to that of N-acetyl-beta-D-glucosaminidase activity with about 12-fold and sixfold increases, respectively . None of the variables returned to baseline by 19 days after virus exposure . Increased elastase activity preceded colonization by P haemolytica and decreasing elastase activity preceded decreasing P haemolytica concentration in the nasal secretions . A causal relation between elastase activity and P haemolytica colonization could be mediated by cleavage of epithelial cell surface fibronectin and exposure of receptors.

Rinsho Byori, 1992 May, 40(5), 547 - 51
{Clinicobacteriological study of Pasteurella multocida infection as a zoonosis . (2)--Comparisons of P . multocida separated from patients and their pet dogs and cats}; Arashima Y et al.; A total of 17 strains of Pasteurella multocida, of which 13 were isolated from patients treated at Nihon University Itabashi Hospital or Nihon University Surugadai Hospital between April, 1984 and March, 1991 and 4 from 1 dog and 3 cats kept by the patients, were evaluated with respect to their biochemical properties, sensitivity to drugs, and serotype . The isolated strains were all considered to be Pasteurella multocida subsp . multocida because of the agreement of their responses to indole, sorbitol and dulcitol with those of this subspecies, except for 1 sorbitol-negative strain of Pasteurella multocida subsp . septica isolated from 1 patient who had been bitten by a cat . All the isolated strains showed high sensitivities to various drugs . The serotype was capsular type A, which is often observed in cats and dogs, in 7 strains, which consisted of 6 of the 7 strains derived from the airway of the patients and 1 of the 6 strains derived from bit or scratch wound . The remaining strains could not be classified . Five morphological types, namely 1, 3, 3.8, 6, and 8 were observed . In 2 patients, Pasteurella multocida subsp . multocida of the same serotype was also isolated from their cats . One of these patients had intimate contact with the cat including kissing . Our findings suggest that: 1) Pasteurella multocida subsp . multocida has been responsible for most conventional cases of Pasteurella multocida infection . 2) Strains isolated from patients differ in the capsular type according to the disease.(ABSTRACT TRUNCATED AT 250 WORDS)

Pathol Biol (Paris), 1992 May, 40(5), 471 - 8
{Antibiotic sensitivity of forty-four strains of group EF4 bacteria: study of minimum inhibitory concentrations using the agar dilution method}; Lion C et al.; EF4 bacteria are found in animal saliva and may contaminate bite wounds . Minimum inhibitory concentrations of 36 antimicrobials against 44 EF4 strains were determined using dilution in Mueller-Hinton agar . EF4 bacteria were susceptible to aminopenicillins, carboxypenicillins, ureidopenicillins, third-generation cephalosporins, fluoroquinolones, rifampicin, and trimethoprime-sulfamethoxazole . Susceptibility was intermediate for penicillin G, low for macrolides and variable for aminoglycosides . EF4 bacteria were resistant to lincomycin and trimethoprime . Routine prophylactic treatment of bite-induced Pasteurella infections using an aminopenicillin or a cycline also protects against EF4 infection of the wound.

Trop Anim Health Prod, 1992 May, 24(2), 97 - 102
An outbreak of haemorrhagic septicaemia (septicaemic pasteurellosis) in cattle in Zimbabwe; Lane EP et al.; An outbreak of haemorrhagic septicaemia caused by Pasteurella multocida in beef cattle in Zimbabwe grazing effluent-irrigated pastures, is described . The outbreak occurred during the wet summer months and predisposing stress factors included excessive rainfall and unusual cold weather during the preceding month . History, clinical features and post-mortem findings were consistent with reports of the disease from other countries, except that meningitis was also a constant feature . Morbidity approached 77% and mortality 5 per cent . Prophylactic treatment and vaccination with a killed bacterin together with a return of warmer and drier weather were probably important in halting the outbreak.

Hear Res, 1992 Apr, 59(1), 1 - 6
Incidence of otitis media in CBA/J and CBA/CaJ mice; McGinn MD et al.; The inbred CBA/J mouse has become a standard experimental animal for auditory study because of its lifelong good hearing . In a newly established mouse breeding colony that housed CBA/J and CBA/CaJ mice to reared as auditory subjects, otitis media frequently afflicted CBA/J mice, reaching an incidence of 90% in animals greater than 400 days of age . Otitis media was not found in CBA/CaJ mice . Three attempts to establish a colony that was free of otitis were unsuccessful . Although the primary pathogen was not clearly established, Pasteurella pneumotropica was isolated from infected bullae . Partial control of otitis media followed the introduction of tetracycline prophylaxis . The CBA/CaJ mice may be suitable replacements for CBA/J mice in studies that require inbred mice with good hearing, since their auditory thresholds did not differ significantly from those of otitis-free CBA/J mice.

Avian Dis, 1992 Apr-Jun, 36(2), 423 - 31
Vaccination of turkeys with cell-free culture filtrate of Pasteurella multocida: effects of ultrafiltration and endotoxin removal; Ficken MD et al.; Cell-free culture filtrate (CCF) of Pasteurella multocida strain R44/6 (serotype 3/4/9/12) was fractionated by ultrafiltration into fractions of less than 10,000, greater than 10,000, greater than 30,000, and 10,000 to 30,000 molecular weight (MW) . The less-than-10,000-MW fraction contained little endotoxin comparable to bacteriologic medium; the 10,000-to-30,000-MW fraction had a moderate amount of endotoxin, whereas the greater-than-10,000- and greater-than-30,000-MW fractions contained high levels of endotoxin . Following ultrafiltration, each fraction, except the less-than-10,000-MW fraction, was divided into two equal parts, and endotoxin was removed from one part . Turkeys were vaccinated with the various MW fractions of CCF, with and without endotoxin, via the air sacs at 6 and 9 weeks of age and compared with negative controls given bacteriologic medium and positive controls vaccinated with a commercial bacterin . Before oral challenge with strain P-1059 (serotype 3) at 12 weeks of age, antibody titers were detected only in positive control turkeys . Protection against challenge, as measured by post-challenge mortality and body-weight gain, was provided by the greater-than-10,000-, greater-than-30,000-, and 10,000-to-30,000-MW fractions containing endotoxin and the commercial bacterin . Turkeys that had been vaccinated with bacteriologic medium and the four different fractions without endotoxin were not protected . Results indicated that endotoxin in CCF of P . multocida is critical in protecting turkeys from pasteurellosis.

Avian Dis, 1992 Apr-Jun, 36(2), 290 - 5
Spondylitis in turkeys associated with experimental Pasteurella anatipestifer infection; Cooper GL et al.; Nine previously vaccinated turkeys were inoculated intravenously with Pasteurella anatipestifer, and blood samples were taken periodically to evaluate the potential of chronically infected turkeys to serve as reservoirs of infection for blood-feeding arthropod vectors . Vertebral osteomyelitis (spondylitis), as yet unreported in the literature in association with infection with the organism, was found in the thoracic vertebrae of five out of nine inoculated turkeys, and P . anatipestifer was isolated from the thoracic vertebrae of three of the five . The organism was isolated from the peripheral blood of six turkeys 24 hours postinoculation and from the peripheral blood of one turkey 7 days postinoculation . The organism was also isolated from the heart blood of two birds at necropsy--from one at 21 days and, following an intramuscular injection of dexamethasone, from the other turkey at 38 days postinoculation.

Avian Dis, 1992 Apr-Jun, 36(2), 272 - 81
Restriction endonuclease analysis of Pasteurella multocida isolates from three California turkey premises; Christiansen KH et al.; Three California turkey premises that had repeated outbreaks of fowl cholera were studied for periods of 2 to 4 years . Using biochemical, serologic, plasmid DNA, and restriction endonuclease analyses of isolates of Pasteurella multocida from turkeys and wildlife on the premises, strains of the organism were found to be enzootic on two of the premises . On the third, a variety of strains of P . multocida were isolated from fowl cholera outbreak flocks.

Avian Dis, 1992 Apr-Jun, 36(2), 262 - 71
Transmission of Pasteurella multocida on California turkey premises in 1988-89; Christiansen KH et al.; Restriction endonuclease analysis (REA) of whole-cell DNA was used to determine possible sources of Pasteurella multocida for each outbreak of fowl cholera occurring in turkey flocks in eight commercial poultry companies in California from October 1988 to September 1989 . Over this period, 179 isolates of P . multocida were obtained from dead turkeys in 80 meat and breeder flocks on 43 premises . P . multocida was isolated from wildlife on five premises . Isolates were characterized by subspecies, serotype, presence of plasmid DNA, and REA type . In 52 (65%) flocks, all isolates of P . multocida had the same REA pattern as the M9 live vaccine strain following digestion of DNA with the restriction enzyme SmaI . Field strains of P . multocida were obtained from 27 (34%) flocks, and one flock (1%) yielded both M9 and a field strain of the organism . REA of field strains of P . multocida revealed 17 different SmaI REA types . Based on matching SmaI REA types, potential sources of P . multocida were identified for 15 of the 28 flocks infected with field strains of the organism, and transmission between turkey premises was a possibility in only seven flocks.

J Comp Pathol, 1992 Apr, 106(3), 221 - 8
The effect of inoculation of Pasteurella haemolytica into the lactating mammary gland of mice, rats, rabbits, sows and cows; Watkins GH et al.; An isolate of Pasteurella haemolytica (A9), which consistently produced severe mastitis in ewes, was inoculated into the lactating mammary glands of a variety of species . Mastitis did not develop after the inoculation of log-phase bacteria into the mammary gland of lactating mice, rats, rabbits or sows but did so in the mammary gland of two cows . Another A9 isolate from a ewe with mastitis and an A1 isolate from a bovine pneumonic lung also induced mastitis in cows . Thus, in this study, P . haemolytica produced mastitis only in ruminant animals.

Can J Vet Res, 1992 Apr, 56(2), 142 - 7
Pulmonary immunity in calves following stimulation of the gut-associated lymphatic tissue by bacterial exotoxin; Bowersock TL et al.; Antibodies in serum and pulmonary lavage fluids were measured in calves following stimulation of the gut-associated lymphatic tissue (GALT) by inoculation of crude leukotoxin of Pasteurella haemolytica into the duodenum through a surgically placed catheter . Nine calves free of P . haemolytica were divided into two groups . Group 1 received an intraduodenal (ID) inoculation of leukotoxin and group 2 received an ID inoculation of phosphate buffered saline . Serum and pulmonary lavage fluids were collected weekly and assayed for antibodies specific to P . haemolytica including immunoglobulin (Ig)G, leukotoxin neutralizing antibodies (LNA), and IgA (lavage fluids only) . The multiplicative increase (over baseline) in each class of antibody titer following ID inoculation of leukotoxin, the composite geometric mean increase of all antibodies together, and the composite number of the five antibody titers which increased at least fourfold were computed . Results showed that the geometric mean of each antibody titer and the two composite indices was higher in the GALT-primed groups than in the sham-primed group . The differences were statistically significant (p less than 0.05) for serum IgG and for the two composite indices . This experiment demonstrates for the first time that GALT stimulation by bacterial exotoxins results in increased pulmonary antibody levels in calves.

Can J Vet Res, 1992 Apr, 56(2), 122 - 6
Cytological findings in bronchoalveolar lavage fluid from feedlot calves: associations with pulmonary microbial flora; Allen JW et al.; Samples obtained by bronchoalveolar lavage (BAL) were used to evaluate pulmonary cytology in 59 feedlot calves with clinical signs of respiratory disease (cases) and 60 clinically normal comparison calves (controls) . Many calves in both case and control groups had inflammatory changes in the lower respiratory tract, as determined by changes in proportions in the BAL differential cell count . Approximately 35% of cases and 40% of controls showed a normal differential cell count . It therefore appeared that the criteria used to select cases for treatment, which were similar to those often used in the field, were poor predictors of lower respiratory tract disease . A positive association was found between an increased proportion of neutrophils in BAL fluid and isolations of Pasteurella multocida and Mycoplasma bovis from BAL fluid.

Am J Vet Res, 1992 Apr, 53(4), 481 - 4
Colonization of the tonsils of calves with Pasteurella haemolytica; Frank GH et al.; Tonsils of 10 calves were inoculated with Pasteurella haemolytica (PH) and the degree of colonization was followed by collecting sequential tonsil . wash specimens . Tonsils were colonized for at least 3 weeks after instillation of PH into the tonsillar sinus . Calves with colonized tonsils responded with serum and nasal secretion antibody responses to PH and to leukotoxin . Pasteurella haemolytica was detected in nasal mucus specimens of 2 calves during the week after inoculation of the tonsils, but all other specimens were culture-negative . Infectious bovine rhinotracheitis virus-induced respiratory tract disease 25 days later did not elicit a population increase of PH in the tonsils, and did not elicit shedding of PH in nasal mucus.

Am J Vet Res, 1992 Apr, 53(4), 472 - 6
Comparison of antigens of Pasteurella haemolytica serotype 1 grown in vitro and in vivo; Confer AW et al.; To determine whether antigenic differences exist in Pasteurella haemolytica serotype 1 grown in different culture conditions, the bacteria was grown on solid enriched medium, in broth culture, and in tissue chambers subcutaneously implanted in the flanks of calves . The organisms obtained by each culture method were comparable with respect to encapsulation and lipopolysaccharide content . In the bacteria grown in vivo, several unique high molecular-mass (greater than 150 kDa) protein antigens were found by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and protein immunoblotting . Bacteria grown in vitro had higher concentrations of a 49- and a 26-kDa protein than the organisms grown in vivo . The concentration of several major proteins (30, 42, 55, 71, and 100 kDa) were similar among the organisms grown by the three cultural conditions . Although the high molecular-mass antigens were unique for the chamber-grown bacteria, they were recognized by serum from a calf that had been vaccinated with formalin-killed, solid medium-grown P haemolytica and were resistant to challenge exposure with the live organism . This recognition of antigens by serum from the P haemolytica-resistant calf that had been vaccinated with solid-medium-grown bacterium, indicates that the high molecular-mass antigens from chamber-grown P haemolytica may be precursors of or share antigenic determinants with other P haemolytica proteins and may not be important for consideration in vaccine formulation.

South Med J, 1992 Apr, 85(4), 442 - 3
Bacteremia due to Pasteurella multocida; Morris JT et al.; Pasteurella multocida should be considered as a possible etiologic agent in any infection that is the result of an animal bite or scratch . Because of its opportunistic capability, it should be included among the possible pathogens in bacteremia, particularly in any patient with immunosuppression or liver cirrhosis, especially if there is a history of animal exposure.

J Infect Dis, 1992 Apr, 165(4), 651 - 7
Alveolar macrophage and neutrophil interactions in Pasteurella haemolytica-induced endothelial cell injury; Sharma SA et al.; Pasteurella haemolytica, the cause of fibrinous pleuropneumonia in cattle, produces extensive microvascular endothelial cell damage . This study investigated endothelial cell-leukocyte interactions by using a Millicell coculture assay system that modeled the bovine pulmonary alveolar defense system and showed that P . haemolytica-mediated endothelial cell damage was enhanced by the presence of alveolar macrophages, presumably due to soluble alveolar macrophage products . The alveolar macrophage-enhanced endothelial cell damage occurred regardless of the presence of anti-P . haemolytica immune serum; however, neutrophils and immune serum effectively prevented endothelial cell damage . These results suggest that alveolar macrophages are ineffective in controlling P . haemolytica growth and actually promote endothelial cell damage.

Infect Immun, 1992 Apr, 60(4), 1401 - 5
Use of restriction endonuclease analysis and ribotyping to study epidemiology of Pasteurella multocida in closed swine herds; Zhao G et al.; One hundred and sixty-four clinical isolates of Pasteurella multocida recovered from two swine herds in Minnesota were characterized by restriction endonuclease analysis (REA) and rRNA gene restriction fragment length patterns . Bacterial DNA was digested with HpaII and electrophoresed in 0.55% agarose . Restriction fragments were transferred by Southern blot to nylon membranes and then hybridized with digoxigenin-dUTP-labeled Escherichia coli rRNA . Four different REA patterns were observed among the 156 serotype A strains isolated from herds A and B . The two most common REA types (1 and 2) represented 92% of the strains analyzed, while REA types 3 and 4 were observed only in lung samples and accounted for 8% of the isolates . Two different ribotypes were observed for these serotype A isolates . Ribotype I consisted of the most common types, 1 and 2, found by DNA fingerprinting . Ribotype II included REA types 3 and 4 . Results from both herds suggest that in closed swine populations, a single strain of P . multocida predominates and causes disease . It is concluded that these genomic fingerprinting techniques were highly discriminatory and that capsular serotyping in combination with REA or ribotyping is an appropriate technique for epidemiological studies of P . multocida of swine origin.

Vet Pathol, 1992 Mar, 29(2), 93 - 103
Bovine focal proliferative fibrogranulomatous panniculitis (Lechiguana) associated with Pasteurella granulomatis; Riet-Correa F et al.; In southern Brazil, cattle are affected by a disease known locally as Lechiguana and characterized by large subcutaneous swellings . Eighteen cases were examined clinically; 17 of the cattle had a single swelling, and one had two swellings . In 14 of the 18 cases, the swellings were located over the scapula and adjacent regions . The subcutaneous masses reached maximum dimensions of 45 x 50 cm, with heights above the skin surface of 5-25 cm . Growth was rapid, often taking place in 15 to 60 days . Histologically, all lesions were focal proliferative fibrogranulomatous panniculitis and consisted of focal proliferation of fibrous tissue that was infiltrated by plasma cells, eosinophils, lymphocytes, and sometimes neutrophils . An eosinophilic lymphangitis was also present, which sometimes resulted in the destruction of the lymphatics and the formation of eosinophilic microabscesses . Small granulomas, sometimes containing radiating clubs, and Splendore-Hoeppli material were present in the regional lymph node . Pasteurella granulomatis was isolated from the subcutaneous masses of 14 of the 18 natural cases . All 11 of these cases recovered following treatment with 3 g of chloramphenicol daily for 5 days . Untreated animals died . Because the area of anatomic distribution is similar to that infested by Dermatobia hominis, we postulate that this insect may transmit the causative agent . In one steer, a subcutaneous injection of P . granulomatis caused a large subcutaneous swelling consisting of interlacing bundles of collagen infiltrated by neutrophils, eosinophils, and some lymphocytes . Microabscesses, but not lymphangitis and granulomas, were detected . In all 11 cattle inoculated either intramuscularly or subcutaneously with P . granulomatis, purulent abscesses were produced at the sites of the injection, and P . granulomatis was recovered from all lesions.

Berl Munch Tierarztl Wochenschr, 1992 Mar 1, 105(3), 87 - 9
{The significance of antibodies to Pasteurella haemolytica A1 in the colostrum of cows and blood serum of calves}; Schimmel D et al.; Antibodies to P . haemolytica were detected in colostral sera of cows and blood sera of calves by means of indirect haemagglutination (IHA) and ELISA . The ELISA titres of the colostral sera of the dams and of the blood sera of the calves showed a significantly positive correlation . There is a positive correlation between the titres of the two serological methods . The results of the challenge infection with P . haemolytica A 1 do not show a correlation between the titres and the severity of the disease . Heifer calves, however, developed more severe pneumonias than calves from older cows.

Appl Environ Microbiol, 1992 Mar, 58(3), 932 - 6
Survival of toxigenic Pasteurella multocida in aerosols and aqueous liquids; Thomson CM et al.; The survival of toxigenic Pasteurella multocida in air and liquids was studied to identify possible risk factors in the etiology of atrophic rhinitis . In aerosols, at low relative humidity (28%), the viability of toxigenic P . multocida 5 min after aerosolization was at least 22% of its initial value . Viability at low relative humidity declined to 8% after 45 min . Viability at high relative humidity (79%) was 69% after 5 min and declined to 2% after 45 min . Survival of toxigenic P . multocida in liquids depended on storage and constituents in the liquid . Toxigenic P . multocida became nonculturable 1 to 14 days after inoculation in water and artificial seawater, depending on the storage temperature . Toxigenic P . multocida stored at 37 degrees C could be detected for up to 6 days in pig slurry and more than 36 days in Bacto Tryptose broth and nasal lavages . However, in Bacto Tryptose broth and nasal lavages stored at 4 degrees C, P . multocida was detected for up to 14 days whereas at 15 and 37 degrees C it was detected for more than 49 days . These results suggest that aerosols and fomites can play a role in the transmission of atrophic rhinitis.

South Med J, 1992 Mar, 85(3), 329 - 30
Septic arthritis due to Pasteurella multocida; Kumar A et al.; We have reported two cases of septic arthritis caused by Pasteurella multocida, the first, septic polyarthritis unrelated to animal contact and the other, septic monarthritis after a cat scratch . Serious infections with this gram-negative bacillus must be treated with intravenous penicillin, ampicillin, or a third-generation cephalosporin; aminoglycosides are not effective . A high index of suspicion is appropriate when there is a history of a recent animal-inflicted wound, though infection may also occur with either nontraumatic animal contact or no recognized animal contact at all.

Res Microbiol, 1992 Mar-Apr, 143(3), 263 - 9
Construction of a broad host range shuttle vector for gene cloning and expression in Actinobacillus pleuropneumoniae and other Pasteurellaceae; Frey J; We have constructed a pair of broad host range expression vectors, pJFF224-NX and pJFF224-XN, based on plasmid RSF1010, which enable cloning and efficient expression of genes in Actinobacillus pleuropneumoniae and Pasteurella haemolytica and in Escherichia coli . The vectors consist of the minimal autonomous replicon of the broad host range plasmid RSF1010 and a type II chloramphenicol acetyl transferase gene for chloramphenicol resistance selection . In addition, they contain a gene expression cassette based on the E . coli bacteriophage T4 gene 32 promoter region and a transcription stop signal, which are separated by a segment of multiple cloning sites in both orientations . Electroporation and subsequent selection for chloramphenicol resistance was used for the introduction of the vectors in A . pleuropneumoniae and P . haemolytica . A promoterless xy/E gene from the Pseudomonas putida TOL plasmid was cloned onto pJFF224-NX . This plasmid enabled efficient expression of active catechol2,3oxygenase in A . pleuropneumoniae and P . haemolytica . It was stably maintained in A . pleuropneumoniae without antibiotic selection, showing less than 0.1% loss after 100 generations, while native RSF1010 and other RSF1010-based vectors were unstable in this host.

J Immunol, 1992 Mar 1, 148(5), 1458 - 64
Cleavage of the cell-surface O-sialoglycoproteins CD34, CD43, CD44, and CD45 by a novel glycoprotease from Pasteurella haemolytica; Sutherland DR et al.; The study of structural/functional characteristics of the cell-surface glycoproteins of leukocytes has led to a better understanding of the differentiation and maturation of hematopoietic cells . We have assessed the ability of a unique metalloprotease that is secreted by the bovine fibrinous pneumonia pathogen Pasteurella haemolytica, to cleave cell-surface glycoproteins expressed on human leukocytes . Biochemical analysis shows that the O-glycosylated cell surface Ag CD34, CD43 (leukosialin), CD44 (hyaluronic acid receptor), and CD45 (leukocyte common Ag), are all cleaved by this protease . Although these enzyme-sensitive structures contain N-linked glycans, they are all extensively glycosylated with O-linked carbohydrates, which are especially abundant on CD34 and CD43 . In contrast, the glycoproteins CD18/11a,b,c (leukocyte integrins), CD71 (transferrin receptor), HLA class I, and 8A3 Ag, which contain N-linked glycans but no O-sialo-glycans, were resistant to the action of the enzyme . Inasmuch as previous studies using glycophorin A had indicated that the substrate specificity of this enzyme may be uniquely restricted to the cleavage of O-sialoglycoproteins, we have designated this activity, P . haemolytica glycoprotease . Immunofluorescence analysis with a variety of antibodies to different epitopes of the P . haemolytica glycoprotease-sensitive structures indicate that this enzyme may have widespread applications in epitope-mapping studies, and represents a novel tool with which to study structure/function relationships for O-sialoglycosylated cell-surface proteins . However, most significantly these results suggest that the P . haemolytica glycoprotease may be of use in the affinity purification and recovery of clinically important leukocyte subsets, such as primitive hematopoietic progenitors that express CD34.

Infect Immun, 1992 Feb, 60(2), 565 - 70
Molecular chimerization of Pasteurella haemolytica leukotoxin to interleukin-2: effects on cytokine and antigen function; Hughes HP et al.; A chimeric recombinant protein composed of the lktA gene product from Pasteurella haemolytica fused to bovine interleukin-2 (IL-2) was made . The LKT-IL-2 chimera was compared with recombinant bovine IL-2 with regard to the ability to induce proliferative responses and LAK cell activity in bovine peripheral blood mononuclear cells in vitro . In both instances, chimerization had no effect on IL-2 activity . Similarly, the LKT component was unaffected in its ability to induce an effective immune response after immunization . The adjuvant properties of IL-2 have been established in a number of models, and this effect was tested by using the chimera . A multiple-injection protocol of LKT-IL-2 was compared with single-dose administration of LKT . The results obtained indicate that while there was no increase in specific antibody production, the IL-2 component of the chimera may be able to affect antigen-specific proliferation, as assessed by limiting-dilution analysis . Use of cytokine-antigen chimeras may provide a valuable antigen-adjuvant formulation that is simple to produce and purify and thus have economic advantages over conventional preparations . Furthermore, chimerization will also ensure that the adjuvant acts at the same site as the antigen, thus optimizing immunostimulatory activity.

Zentralbl Veterinarmed B, 1992 Feb, 39(1), 10 - 8
Further investigations on Pasteurella multocida infections in feral birds injured by cats; Korbel R et al.; A total of 64 Pasteurella multocida strains (46 out of 11 different feral bird species, partly injured by cat bites, and 18 strains originating from clinically healthy cats) were biochemically differentiated . As a result, 67.4% of the strains from feral birds and 61.1% from the cats were classified as the subspecies multocida, whilst 21.7% and 27.8% were identified as the subspecies septica . The percentage of frequency for both the subspecies was of a comparable order of magnitude from the birds injured by cat bites and from cats (58.6% and 61.1%, 24.1% and 27.8% resp.), whereas the frequency from other feral birds differed considerably (82.4% and 17.6%) . Maltose-positive strains were only demonstrable in birds with wounds inflicted by cats . To date, maltose-positive strains have only been obtained from one cat and one human being with an injury caused by a cat . The results of this investigation confirm the possibility of the direct transmission of Pasteurella multocida via cat bites . 19 strains from feral birds and 15 strains from cats were tested for their capability to produce toxins . The results of these tests were negative . The present paper also describes the pathologic-anatomical and histopathological lesions caused by the infection in feral birds.

Zentralbl Bakteriol, 1992 Feb, 276(3), 366 - 73
Cell surface characteristics and virulence in mice of Pasteurella multocida; Dubreuil JD et al.; The virulence of three avian strains of Pasteurella multocida was evaluated in mice . Strains P-1059I (serotype A:3) and its uncapsulated variant P-1059B and strain 2723 (serotype A:16) were compared . Capsular material thickness after polycationic ferritin labelling of dextrose starch agar (DSA)-grown P . multocida was shown to vary with the strain and was not always related to virulence . Addition of alpha,alpha' bipyridyl (BIP) (160 microM) to the culture medium did not affect capsule production but increased virulence of strains P-1059B and 2723 . None of the strains tested showed dermonecrotic activity . Using sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE), outer membrane protein (OMP) profiles indicated for strains P-1059I and P-1059B three proteins of 30, 35, and 38 KDa with the 30 KDa protein being the major one . Strain 2723 showed the same OMP profile but the 38 KDa protein was the major one . DSA + BIP-grown strains showed the same OMP profiles . Whole cell profiles were similar for all strains tested . However, addition of BIP to the culture media increased the virulence of strains P-1059B and 2723 and for all strains a 39 KDa protein was induced by the iron chelator . The results indicate that encapsulation may be important for virulence, but other surface components such as OMPs may be required as well.

Dtsch Tierarztl Wochenschr, 1992 Feb, 99(2), 63 - 6
{Detection of the nucleolar activity in alveolar macrophages of swine}; Kohler H; With the help of the toluidine blue-staining-technique different characteristic shapes of alveolar macrophage nucleoli were detected, indicating differences in the rRNA synthesis activity of these cells . After application of a Pasteurella multocida aerosol the percentage of alveolar macrophages with a high rRNA synthesis increased . No correlation between the nucleolar activity and the efficacy of the clearance of aerogenously administered Pasteurella could be measured.

Clin Infect Dis, 1992 Feb, 14(2), 497 - 500
In utero infection due to Pasteurella multocida in the first trimester of pregnancy: case report and review; Waldor M et al.; To our knowledge, we report the first case of in utero infection in humans due to Pasteurella multocida in the first trimester of pregnancy . After antibiotic therapy was initiated, the mother recovered rapidly from bacteremia but the fetus was spontaneously aborted . Pathological examination of the fetus and placenta revealed acute deciduitis and chorionitis, indicating an infectious etiology for the abortion . The clinical and pathological features of this case are compared with those of the five previously reported cases of P . multocida bacteremia in humans that occurred during pregnancy.

Kansenshogaku Zasshi, 1992 Feb, 66(2), 232 - 5
{The first report of human chronic sinusitis by Pasteurella multocida subsp . multocida in Japan}; Arashima Y et al.; A 53-year-old male visited our hospital due to nasal obstruction persisting for 6 months and constant rhinorrhea . Pasteurella multocida subsp . multocida was isolated from his nasal discharge and lavage fluid of the maxillary sinus, and also from the oral cavity of the dog he kept . The bacterial strains isolated from the patient and dog were identical in terms of biochemical properties, and drug sensitivity . Although serotype was different, the strain from the patient showed (A:6) and that from his dog showed (A:5) . The microorganism is not present in the general environment . The patient had contact with his dog such as he kissed it frequently, gave it food with his chopsticks et al. . From the mouth of the people who kiss one's dog, we detected Pasteurella of the same character of bacteria as from the mouth of the dog . We detected two Pasteurella multocida of different character from only one mouth of a cat . Pasteurella multocida was checked in only one colony for sero type . Sero type A is the popular type for dogs and cats . The above suggest that their was a high possibility that the Pasteurella multocida subsp . multocida found in the patient was from his dog . In Japan, the incidence of Pasteurella multocida subsp . multocida infection has been increasing . In 1969, the Japanese Ministry of Health and Welfare officially communicated this infection as a zoonosis to related institutions . At both medical and surgical departments, wither the patient keeps a pet should be confirmed during interview, and guidance of pet keeping methods is important in some cases from the aspect of clinical bacteriology.(ABSTRACT TRUNCATED AT 250 WORDS)

Kansenshogaku Zasshi, 1992 Feb, 66(2), 221 - 4
{Clinicobacteriological study of Pasteurella multocida as a zoonosis (1) . Condition of dog and cat carriers of Pasteurella, and the influence for human carrier rate by kiss with the pets}; Arashima Y et al.; Pasteurella multocida is a gram-negative short rod-shaped bacteria, which is a part of the indigenous flora of the oral cavity of many animals other than man . The number of reports on cases of infections with this bacterium due to animal bites and/or scratches, bacterial infections of the respiratory tract, sepsis due to this organism and death caused by the bacteria have been increasing in recent years . We investigated P . multocida in the hair and oral cavity of 3 dogs and 29 cats according to the classification of Mutters et al. . We also studied the relationship between the carrier rate for Pasteurella in the oral cavity and kissing of pets in 24 pet owners (3 dogs and 11 cats) . No P . multocida was isolated from the hair of neither dogs nor cats . One strain of P . multocida subsp . multocida and two strains of P . stomatis, were isolated from the oral cavity of dogs, and 35 strains of Pasteurella were isolated from the oral cavity of cats . Two strains of P . multocida subsp . multocida, whose biochemical properties were different, were detected in the oral cavity of one cat . In three cats, Pasteurella other than P . multocida subsp . multocida was isolated from the same oral cavity . No Pasteurella was detected in the oral cavity of 19 pet owners who had not kissed their cats, whereas P . stomatis was isolated from the oral cavity of one of 2 pet owners who had kissed their cats and in 2 of 3 pet owners who had kissed their dogs (the same bacteria was isolated from a dog that was being kept by some of these positive pet owners).(ABSTRACT TRUNCATED AT 250 WORDS)

J Gen Microbiol, 1992 Feb, 138 ( Pt 2), 283 - 8
Antigenic analysis of iron-regulated proteins in Pasteurella haemolytica A and T biotypes by immunoblotting reveals biotype-specific epitopes; Murray JE et al.; The antigenic relationships of the iron-regulated proteins (IRPs) in Pasteurella haemolytica A and T biotype strains were examined by SDS-PAGE and immunoblotting . P . haemolytica cells of the A biotype, grown under conditions of iron-limitation, expressed two IRPs, of 35 and 70 kDa . All T biotype strains expressed IRPs with slightly different molecular masses of 37 and 78 kDa . Immunoblotting of all 16 P . haemolytica serotypes was carried out using a panel of polyclonal and monoclonal antibodies raised against serotype A2 antigens . Polyclonal antibodies revealed inter-serotype cross-reactivity towards the 35 and 70 kDa IRPs within the A biotype but no cross-reactivity against a T biotype protein in the 78 kDa region . Monoclonal antibody against the 35 kDa antigen reacted only with the A biotype 35 kDa IRP . Identical profiles were obtained for 10 field isolates of serotype A2, further emphasizing the antigen conservation within the A biotype . These findings reinforce the view that the A and T biotypes of P . haemolytica should be considered as separate species and suggest that IRPs from single A and T biotype strains incorporated into a vaccine might provide cross-protection against all P . haemolytica serotypable strains . Similar studies on the IRPs of 10 untypable strains revealed some of these to have different antigenic reactivities from those observed within the A and T biotypes.

Lab Anim Sci, 1992 Feb, 42(1), 13 - 8
Colonization of rabbits by Pasteurella multocida: serum IgG responses following intranasal challenge with serologically distinct isolates; DeLong D et al.; Enzyme-linked immunosorbent assays (ELISAs) and immunoblots were used to measure serum IgG responses in rabbits which were intranasally challenged with Pasteurella multocida . The responses to two serologically distinct isolates (isolate 1, serotype 3:A and isolate 10, serotype 1:D) were compared and then correlated with the ability of the isolates to colonize the nasal passages . Five rabbits were challenged with each isolate (10(5) CFU); nasal washings and sera were collected weekly for 8 weeks . Serum IgG levels were measured by ELISA and immunoblots, using bacterial whole cells and lipopolysaccharides (LPSs) as antigens . The serum IgG response to isolate 1 was evident earlier and was significantly stronger than the response to isolate 10 (P less than 0.025) . Immunoblots supported this observation and confirmed that both isolates elicited antibodies which reacted with bacterial protein and LPS antigens, with antibody to protein detectable before antibody to LPS . Results of weekly nasal cultures suggested that the antibody response data could be explained by a difference in the ability of the isolates to colonize the nasal passages: isolate 1 was recovered from four of five rabbits for 8 weeks, whereas isolate 10 was recovered for a maximum of 2 weeks, even when the challenge dose was increased tenfold . The strong response elicited by isolate 1 was therefore probably a result of persistent colonization, whereas the weak response to isolate 10 may have resulted from an inability to persistently colonize the nasal passages . The results of this study demonstrate that isolates of P . multocida elicit antibody responses of differing intensities and vary in their ability to colonize the nasal passages.(ABSTRACT TRUNCATED AT 250 WORDS)

Vet Immunol Immunopathol, 1992 Jan 31, 30(4), 341 - 57
Release of tumor necrosis factor-alpha from bovine alveolar macrophages stimulated with bovine respiratory viruses and bacterial endotoxins; Bienhoff SE et al.; The release of tumor necrosis factor-alpha (TNF-alpha) from cultured bovine alveolar macrophages (BAM) was evaluated following stimulation of BAM with bovine herpesvirus-1 (BHV-1), parainfluenza-3 (PI-3) virus, bovine respiratory syncytial virus (BRSV), Escherichia coli 0111:B4 endotoxin, Pasteurella haemolytica type 1 endotoxin, Pasteurella multocida endotoxin, and virus/endotoxin combinations . A cytotoxic assay system using Georgia bovine kidney cells as targets was used to measure TNF-alpha activity . The cytotoxic activity was neutralized by an anti-human TNF-alpha monoclonal antibody . Stimulation of BAM with 1 median tissue culture infectious dose (TCID50) of live or ultraviolet (UV)-inactivated PI-3 virus/cell resulted in release of TNF-alpha in significantly (P less than 0.05) higher amounts than sham-induced BAM . The quantities of TNF-alpha released after live or UV-inactivated BHV-1 or BRSV induction were not significantly higher than sham-induced BAM . E . coli 0111:B4, P . haemolytica type 1 and P . multocida endotoxins stimulated TNF-alpha release in a dose-dependent manner . Sequential exposure of BAM to 1 TCID50 per cell of either live BHV-1, PI-3 virus or BRSV and then 5 micrograms ml-1 of either E . coli 0111:B4, P . haemolytica type 1 or P . multocida endotoxin caused a significant (P less than 0.05) reduction in detectable TNF-alpha in seven of nine virus/endotoxin combinations tested, when compared with 5 micrograms ml-1 of endotoxin alone . Parainfluenza-3 virus/endotoxin combinations stimulated higher TNF-alpha release when compared with other virus/endotoxin combinations . Five out of six test animals had serum-neutralizing antibodies to PI-3 virus, one out of six had serum-neutralizing antibodies to BHV-1, and two out of six had serum-neutralizing antibodies to BRSV, suggesting a possible relationship between serum neutralizing antibodies and TNF-alpha release from in vitro cultivated BAM.

Tijdschr Diergeneeskd, 1992 Jan 15, 117(2), 35 - 7
{Pasteurella haemolytica serotypes in cattle}; Visser IJ et al.; Eight-two bovine Pasteurella haemolytica strains were serotyped . The majority of the strains were isolated from calves which had died from fibrinous pneumonia and small numbers from cases of pleuritis, sepsis and abortion . A total of eight different serotypes were noticed . Serotype A1 was found to be the most prevalent 37.8 per cent, followed by serotype A2 with 20.7 per cent . Antibiotic resistance was found for sulfonamide, tetracycline and penicillin in about half of all strains . Conventional combination of biotype to serotype per strain was not confirmed by the results of arabinose and trehalose fermentation.

Toxicol Pathol, 1992, 20(1), 103 - 11
Light microscopic and ultrastructural pathology of seminiferous tubules of rats given multiple doses of Pasteurella multocida group D protein toxin; Ackermann MR et al.; Male Holtzman rats were given subcutaneous doses of a purified Pasteurella multocida group D heat-labile toxin on alternate days for up to 22 days . Rats were necropsied at 18 days or 36 days (14 days after last dose of toxin) or when moribund, and testicles were taken for histologic and ultrastructural examination . Other selected tissues, including liver and spleen, were taken for histologic examination . Histologically, testicular and splenic lesions occurred more consistently and at much smaller doses when compared with lesions in other target organs such as liver . Testicular and splenic lesions were present in all rats (6/6) given 0.8 micrograms/kg toxin and were seen in some rats (1/6) given as little as 0.2 micrograms/kg toxin . Only 3/6 rats given 0.8 micrograms/kg toxin had hepatic lesions; no hepatic lesions were seen at doses of 0.2 micrograms/kg . Testicles from toxin-treated rats were smaller and weighed less than controls . Seminiferous tubules were moderately dilated and lined by polygonal sertoli cells . The normal spermatogenic maturation sequence and mature spermatids were absent, and many tubules contained multinucleate spermatocytes . Severely affected tubules were necrotic and mineralized . Ultrastructurally, there was necrosis of adluminal spermatocytes, multinucleate cell formation, and spaces between Sertoli cell plasma membranes . Testicular lesions were similar to those described for vitamin D-deficient rats, vitamin A-deficient rats, vasectomized rats, and rats given intravenous tumor necrosis factor; however, rats given lethal doses of toxin did not have elevated levels of TNF alpha activity.

J Bacteriol, 1992 Jan, 174(1), 291 - 7
Separable domains define target cell specificities of an RTX hemolysin from Actinobacillus pleuropneumoniae; McWhinney DR et al.; The leukotoxin (LktA) from Pasteurella haemolytica and the hemolysin (AppA) from Actinobacillus pleuropneumoniae are members of a highly conserved family of cytolytic proteins produced by gram-negative bacteria . Despite the extensive homology between these gene products, LktA is specific for ruminant leukocytes while AppA, like other hemolysins, lyses erythrocytes and a variety of nucleated cells, including ruminant leukocytes . Both proteins require activation facilitated by the product of an accessory repeat toxin (RTX) C gene for optimal biological activity . We have constructed six genes encoding hybrid toxins by recombining domains of ltkA and appA and have examined the target cell specificities of the resulting hybrid proteins . Our results indicate that the leukocytic potential of AppA, like that of LktA, maps to the C-terminal half of the protein and is physically separable from the region specifying erythrocyte lysis . As a consequence, we were able to construct an RTX toxin capable of lysing erythrocytes but not leukocytes . The specificity of one hybrid was found to be dependent upon the RTX C gene used for activation . With appC activation, this hybrid toxin lysed both erythrocytes and leukocytes, while lktC activation produced a toxin which could attack only leukocytes . This is the first demonstration that the specificity of an RTX toxin can be determined by the process of C-mediated activation.

Infect Immun, 1992 Jan, 60(1), 56 - 62
A neutral glycoprotease of Pasteurella haemolytica A1 specifically cleaves O-sialoglycoproteins; Abdullah KM et al.; A neutral metalloprotease with marked specificity for an O-sialoglycoprotein has been isolated from culture supernatants of Pasteurella haemolytica A1 . The 35-kDa enzyme cleaves human erythrocyte glycophorin A, which is O glycosylated, but does not cleave N-glycosylated proteins or nonglycosylated proteins . Glycophorin A was cleaved when it was present in situ in erythrocyte ghost plasma membranes or when it was free in solution . The glycoprotease did not hydrolyze glycophorin A from which sialate residues had been removed by neuraminidase treatment . An immobilized preparation of the enzyme cleaved glycophorin A at several positions, with a major site of cleavage at Arg-31-Asp-32 . The glycoprotease is inhibited by EDTA, citrate, and ascorbate, but inhibition appears to be due to the masking of metal ion activators rather than to their removal . The enzyme is not inhibited by phosphoramidon, an inhibitor of other bacterial neutral metalloproteases.

Avian Dis, 1992 Jan-Mar, 36(1), 97 - 100
Comparative susceptibility of caponized and uncaponized tom turkeys to Pasteurella multocida; Friedlander RC et al.; When susceptibility to virulent Pasteurella multocida was compared, there was no significant (P greater than 0.05) difference between caponized and uncaponized tom turkeys . Neither was there any significant (P greater than 0.05) difference between the surviving caponized and uncaponized toms in the development of serum anti-P . multocida antibody . However, at 28 weeks of age, the average live body weight of the caponized toms was significantly (P less than 0.05) lower than that of the uncaponized toms . Turkeys were caponized when 9 weeks old, and different groups were exposed to P . multocida when 13, 18, 23, and 28 weeks old.

Microb Pathog, 1992 Jan, 12(1), 63 - 8
Conditions for transformation of Pasteurella multocida by electroporation; Jablonski L et al.; Conditions for electroporation of plasmid DNA into Pasteurella multocida were determined for use in developing a cloning system to study virulence factors of P . multocida . The highest efficiency of transformation (1.25 x 10(7) cfu/micrograms DNA) was obtained when 7.6 x 10(10) cells of P . multocida strain R473 were electroporated at 12.5 kV/cm (10 ms, 5 ng of pVM109) . Transformation efficiencies of cells prepared at mid-log-phase were approximately 0.5 log10 lower than early, late, or stationary phases . Neither pBR322 nor pUC-19 were able to transform strain R473 under these conditions, even when DNA concentrations were increased to 1 microgram . When pBR322 was ligated with a Pasteurella plasmid, pLAR-1, the hybrid was able to transform strain R473 at an efficiency between 4.5 x 10(2) and 8 x 10(4) cfu/micrograms DNA . Six strains of P . multocida including serotypes A, B, D, and E were transformed successfully.

J Comp Pathol, 1992 Jan, 106(1), 9 - 14
The effect of the intra-mammary inoculation of lactating ewes with Pasteurella haemolytica isolates from different sources; Watkins GH et al.; Lactating Welsh Mountain ewes were inoculated, 3 weeks after lambing, with between 1000 and 10,000 colony forming units of a number of isolates of Pasteurella haemolytica . Isolates from severe, acute mastitis in a ewe, from ovine and bovine pneumonic lesions and from the nasal cavity of healthy lambs, all gave rise to severe, acute mastitis that was clinically indistinguishable from that seen naturally . Two isolates from the milk of ewes with subclinical mastitis did not cause clinical disease after inoculation and, in most ewes, were immediately eliminated . These results suggest that a variety of strains of P . haemolytica are capable of causing severe mastitis in sheep, regardless of their origin, and that there are strains of lower pathogenicity for the mammary gland which are not capable of causing clinical mastitis.

J Vet Intern Med, 1992 Jan-Feb, 6(1), 11 - 22
Pasteurella haemolytica A1 and bovine respiratory disease: pathogenesis; Whiteley LO et al.; The severe fibrinonecrotic pneumonia associated with pneumonic pasteurellosis usually results from colonization of the lower respiratory tract by Pasteurella haemolytica biotype A, serotype 1(A1) . Despite recent research efforts, the authors lack a detailed understanding of the interactions and host response to P . haemolytica in the respiratory tract . The authors hypothesize that management and environmental stress factors or viral infection alters the upper respiratory tract (URT) epithelium allowing P . haemolytica to colonize the epithelium . Once the URT is colonized, large numbers of organisms enter the lung where they interact with alveolar macrophages . Endotoxin, released from the bacteria, crosses the alveolar wall where it activates pulmonary intravascular macrophages, endothelium, neutrophils, lymphocytes, platelets, complement, and Hageman factor leading to complex interactions of cells and mediators . It is the progression of this inflammatory response with neutrophil influx that is ultimately responsible for the pulmonary injury . Leukotoxin is a major virulence factor of P . haemolytica that allows it to survive by destroying phagocytic cells . At subcytolytic concentrations it may also enhance the inflammatory response by activating cells to produce mediators and release reactive oxygen metabolites and proteases.

Avian Dis, 1992 Jan-Mar, 36(1), 84 - 7
Fowl cholera epornitic: antigenic characterization and virulence of selected Pasteurella multocida isolates; Rhoades KR et al.; An epornitic of fowl cholera involving turkey flocks of several farms within a 15-mile radius in Utah was studied . Pasteurella multocida strains isolated from birds in affected flocks were antigenically characterized as A:1, A:3, and B:4, based on capsular sero-grouping and somatic serotyping results . Experimental exposure of poults with each of two strains representing the rarely reported capsular group B indicated that both were virulent.

Scand J Immunol Suppl, 1992, 11, 75 - 80
Evidence of immunosuppression by bovine respiratory syncytial virus; Woldehiwet Z et al.; Respiratory syncytial virus (RSV) is a major respiratory pathogen in human infants and calves . Calves and lambs infected with bovine RSV show mild clinical signs but they are more susceptible to secondary infection with Pasteurella haemolytica . Lambs infected with P . haemolytica 6 days after experimental infection with bovine RSV had significantly greater magnitudes of fever, higher disease and lesion scores and higher mortality rates than those infected with P . haemolytica or bovine RSV alone (P less than 0.05) . Experimental infection with bovine RSV is characterized by alterations in lymphocyte subpopulations and down-regulation of some of their functions . For example, the number of T helper cells is significantly reduced during the first week of infection and peripheral blood mononuclear cells obtained from bovine RSV-infected lambs were less responsive to the mitogen phytohaemagglutinin but more susceptible to P . haemolytica cytotoxin than those obtained from control lambs . Infection with bovine RSV does not significantly affect the humoral immune responses of lambs against P . haemolytica cytotoxin . Bovine RSV does not appear to affect the capacity of alveolar macrophages to present antigens in vitro.

Vet Res Commun, 1992, 16(2), 97 - 105
Passive protection of mice with antiserum to neuraminidase from Pasteurella multocida serotype A:3; Ifeanyi FI et al.; Antiserum to a partially purified neuraminidase from Pasteurella multocida, type A:3, was adsorbed with protease-digested P . multocida type 3 lipopolysaccharide (LPS) to remove LPS immunoreactivity . The LPS-adsorbed antineuraminidase caused a 77% reduction in the neuraminidase activity of homologous P . multocida in an in vitro enzyme neutralization test . All 14 mice passively immunized with the adsorbed antineuraminidase were protected against challenge infection with homologous P . multocida in a mouse protection test . Ten out of 14 mice in one group that received antisera containing antibodies to both neuraminidase and LPS were protected . In contrast, only 1 out of 14 mice that were immunized with pre-immune serum survived the challenge . These results suggest that antiserum to P . multocida neuraminidase was, at least partly, responsible for the protection observed in this study . Neuraminidase may be one of the immunogenic protective proteins present in aqueous extracts of Pasteurella multocida.

Ann Fr Anesth Reanim, 1992, 11(1), 103 - 4
{Pulmonary pasteurellosis in a young patient with multiple trauma}; Thomas C et al.; A case is reported of an 18-year-old male patient who had a road traffic accident, with head and chest injuries . The patient was admitted to the surgical intensive care unit 24 h later because of an alteration of his level of consciousness . He required artificial ventilation . Five days later, he developed right-sided lower lobe pneumonia, treated with positive end-expiratory pressure . A Gram negative organism was found on bronchial brushing, but not in haemocultures . It was identified as Pasteurella multocida, sensitive to beta-lactamines, but not to amikacin . Cefotaxime, which had been started immediately after the arrival of the Gram stain result, was continued . Artificial ventilation was discontinued on day 12, and the patient left the unit on day 15 . The patient was probably a P . multocida carrier, being in close contact with animals before his accident . This bacteria is often found in infected animal bite wounds . Pneumonia due to this bacteria usually occurs in immunodepressed patients, which was not the case here.

Parasitol Res, 1992, 78(6), 525 - 8
Interaction between Ascaris suum and Pasteurella multocida in the lungs of mice; Tjornehoj K et al.; In an experiment including 8 groups of 15 mice, the effect of migrating Ascaris suum larvae in the lungs on the establishment and pathogenicity of aerosol exposure to Pasteurella multocida was investigated . Following aerosol exposure to P . multocida, mice with migrating A . suum in their lungs developed more severe pneumonia and septicaemia than did parasite-free mice . The parasite-induced effect on bacterial pathogenicity was more marked for a non-toxin-producing P . multocida as compared with a toxin-producing strain of P . multocida, possibly due to the higher spontaneous pathogenicity of the non-toxigenic strain of P . multocida . The present results should encourage controlled experiments on possible interactions between A . suum and various airborne microbial infections in pigs.

Vet Res Commun, 1992, 16(3), 177 - 83
Co-migrating and shared antigens of selected Pasteurella haemolytica untypable strains; Simons KR et al.; Bacterial cell envelope preparations from eight untypable strains of Pasteurella haemolytica were compared by sodium dodecyl sulphate-polyacrylamide gel electrophoresis and immunoblotting with rabbit antisera prepared against the eight untypable strains (one untypable strain per rabbit) and with cattle antisera prepared against P . haemolytica serotypes 1, 2, 5, 6, 9 and against one heterologous untypable strain . Numerous comigrating and shared antigens were recognized by the eight rabbit antisera and the P . haemolytica serotype cattle antisera . Comigrating antigens at 43 and 30 kilodaltons (kDa) were recognized by all eight rabbit antisera . Shared antigens, detected by all eight rabbit antisera when reacting against P . haemolytica serotype 1, were recognized at 43, 32, 30, 20 and 15 kDa.

Scand J Infect Dis, 1992, 24(4), 453 - 6
Severe Pasteurella multocida infections in pregnant women; Rollof J et al.; We report 2 cases of severe infections due to Pasteurella multocida, both occurring during pregnancy in previously healthy women . Both women had contact with animals (dog and cat) but neither of them had been bitten . Apart from a slight decrease in IgG levels, no immunological defects could be detected . Both women had received oral phenoxymethylpenicillin in the early phase of the disease, but still fell ill with severe infections . One woman had meningitis while the other suffered from cellulitis with deep abscess formation.

DNA Seq, 1992, 3(2), 89 - 97
Characterization of plasmids with antimicrobial resistant genes in Pasteurella haemolytica A1; Chang YF et al.; Two R plasmids, pYFC1 and pYFC2, from Pasteurella haemolytica A1 encoding sulfonamide, streptomycin (pYFC1), and ampicillin (pYFC2) resistances have been characterized by restriction endonuclease digestions, subcloning or DNA sequencing . pYFC1 consists of 4225 bp and is 51.9% in AT content . Physical mapping indicated a highly conserved region of restriction sites among pYFC1, RSF1010, pGS05, pFM739, pHD148 and pGS03B . pYFC1 encoded a dihydropteroate synthase (29.8 kDa), and streptomycin kinase (29.6 kDa) which is homologous in nucleotide sequences or deduced amino acid sequence to that encoded by a broad-host range IncQ plasmid RSF1010 . Based on the primary structure of pYFC1, the sulfonamide and streptomycin genes are derived from the same ancestor of RSF1010 . pYFC2 is similar to the plasmid from P . haemolytica LNPB51 isolated in France by partial restriction enzyme mapping . pYFC1 and pYFC2 have the same size of 4.2 kbp.

Vet Res Commun, 1992, 16(6), 429 - 36
Controlling tick infestations and diseases in sheep by pour-on formulations of synthetic pyrethroids . A field study; Hardeng F et al.; The use of synthetic pyrethroids in pour-on formulations reduced tick infestations and the incidence of tick-associated diseases in lambs more than dipping in organophosphate acaricides . Though the use of pyrethroids did not prevent the lambs from being infected with tick-borne fever (TBF), the incidence of lambs with lameness (tick pyaemia) or lambs suddenly found dead (Pasteurella haemolytica septicaemia), which often are seen in association with TBF, was reduced . The use of pyrethroids for three years did not seem to affect the prevalence of TBF.

FEMS Microbiol Lett, 1991 Dec 15, 69(1), 23 - 8
Optimal conditions for the analysis of Pasteurella haemolytica lipopolysaccharide by sodium dodecyl sulphate-polyacrylamide gel electrophoresis; Davies RL et al.; The optimal conditions for the analysis of the lipopolysaccharide (LPS) of two serotype A1 isolates and a serotype A2 isolate of Pasteurella haemolytica by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and silver staining were determined . The LPS of the A1 isolates possessed O side chains, consisting of high molecular mass bands with the appearance of a ladder-like pattern, as well as a low molecular mass core-oligosaccharide region; the LPS of the A2 isolate consisted only of the core-oligosaccharide region . Furthermore, the LPS of the two A1 isolates differed in the core-oligosaccharide region . Optimal resolution of low molecular mass LPS components was obtained in a 15% acrylamide resolving gel containing 4 M urea whereas optimal resolution of high molecular mass components was obtained when urea was omitted . Conventional silver staining resulted in excellent visualisation of LPS bands, whereas a modified staining method did not detect additional bands, as has been demonstrated with the LPS of Pseudomonas aeruginosa . Proteinase K digestion of outer membranes gave more clearly defined LPS profiles than did similar digestions of whole cells, and more closely resembled the profiles of purified LPS . With the exception of slight variation in the average molecular mass of a group of O side chains between logarithmic and stationary phases there were no differences in LPS profiles at various stages of the growth cycle; freezing and thawing of LPS samples had no effect on the profiles.

Infect Immun, 1991 Dec, 59(12), 4570 - 5
Protective role of bovine neutrophils in Pasteurella haemolytica-mediated endothelial cell damage; Breider MA et al.; The purpose of this study was to determine if Pasteurella haemolytica can directly injure bovine pulmonary endothelial cells (EC) and if neutrophils have a beneficial or detrimental role in bacterium-EC interaction . Various combinations of live P . haemolytica, heat-killed P . haemolytica, anti-P . haemolytica immune serum, polymyxin B, and bovine neutrophils were added to confluent monolayers of bovine EC . Monitoring included determination of 51Cr release from EC, phase microscopy, and transmission electron microscopy . Although toxic changes were not evident at 5 h postinoculation, both live and heat-killed P . haemolytica produced extensive EC damage by 22 h postinoculation . Damage by live P . haemolytica was prevented only when both neutrophils and immune serum were used . Polymyxin B effectively prevented the toxic effect of heat-killed P . haemolytica, suggesting that lipopolysaccharide was the major toxic factor . Morphological studies showed close apposition of P . haemolytica to EC membranes, neutrophil activation, and adherence to EC but no evidence of neutrophil-associated EC membrane damage . These studies demonstrate that neutrophils and immune serum in combination are effective in preventing EC damage mediated by live P . haemolytica.

Infect Immun, 1991 Dec, 59(12), 4517 - 23
The outer membrane of Pasteurella multocida 3:A protects rabbits against homologous challenge; Lu YS et al.; The protective efficacy of a vaccine purified from the Pasteurella multocida 3:A outer membrane (OM) was evaluated in rabbits by homologous challenge . Twenty-seven rabbits were divided into four groups: 1, vaccinated with OM and challenged; 2, nonvaccinated and challenged; 3, vaccinated with OM only; and 4, nonvaccinated and not challenged . Rabbits were immunized intranasally with 1 mg of OM protein on days 0, 7, 14, and 35, challenged intranasally on day 49, and killed on day 63 . Mortality rates were 0, 67, 0, and 0% for groups 1 through 4, respectively . The prevalence of pneumonia was reduced from 73 (group 2) to 20% (group 1) . The severity of pneumonia was reduced from 0.62 (group 2) to 0.07 (group 1), as measured by the group lesion index . The number of P . multocida in nasal cavities was reduced from 3.89 x 10(5) (group 2) to 6.19 x 10(2) (group 1) . The geometric mean number of P . multocida in lungs was 8,360,000-fold less in group 1 than in group 2 . Similarly, the prevalence of P . multocida colonization in nonrespiratory organs was reduced from 47 (group 2) to 4% (group 1) . Furthermore, group 1 and 3 rabbits developed significantly elevated immunoglobulin A antibodies in nasal secretions and lung lavages and significantly elevated immunoglobulin G antibodies in lung lavages and sera . In addition, rabbit immune sera contained antibodies against P . multocida OM proteins and lipopolysaccharides and inhibited P . multocida proliferation in mouse lungs . These results indicate that a vaccine prepared from the OM of P . multocida provides a significant protection in rabbits against homologous challenge.

Infect Immun, 1991 Dec, 59(12), 4497 - 504
Cytolysins of Actinobacillus pleuropneumoniae serotype 9; Smits MA et al.; Cytolysin I (ClyI) and cytolysin II (ClyII), which are present in the culture supernatant of Actinobacillus pleuropneumoniae serotype 9, are thought to play an important role in the pathogenesis of pig pleuropneumonia . The purpose of this study was to clone and characterize the genetic determinants of these cytolysins . Cloning was accomplished by the screening of DNA libraries for the presence of cytolytic activity and for the presence of DNA sequences homologous to leukotoxin DNA of Pasteurella haemolytica . Both genetic determinants were found to be members of the RTX cytotoxin family . The ClyII determinant was characterized in more detail . It appeared that ClyII more closely resembled the leukotoxin of P . haemolytica than the alpha-hemolysin of Escherichia coli . The ClyII amino acid sequence was identical to a hemolysin gene sequence of A . pleuropneumoniae serotype 5; this finding indicates that the latter gene also codes for ClyII and not for ClyI, as has previously been suggested . The genetic organization of the ClyII determinant differed from the genetic organization of other RTX determinants . Genes responsible for secretion of ClyII were not contiguous with the toxin gene . Instead, secretion genes were present elsewhere in the genome . These secretion genes, however, belong to the ClyI operon . This indicates that the secretion genes of the ClyI operon are responsible for secretion of ClyI and ClyII.

J Anim Sci, 1991 Dec, 69(12), 4876 - 82
Effects of a zinc-deficient diet on tissue zinc concentrations in rabbits; Bentley PJ et al.; Young male New Zealand White rabbits given a diet containing 2 ppm of Zn (Zn-deficient diet) ceased to grow after 5 wk . Control rabbits given diets containing 80 or 85 of ppm Zn and experimental animals given 7 ppm of Zn (low-Zn diet) grew normally . The rabbits given the Zn-deficient diet also exhibited alopecia, skin lesions, and frequent pasteurella infections . These conditions were not observed in rabbits fed the other diets . The testes and thymus were smaller in the rabbits fed the Zn-deficient diet than in rabbits fed the control diet . Serum Zn concentrations in rabbits given the low- or Zn-deficient diets reached new lower levels after 2 wk, and these concentrations were maintained for up to 12 wk . The serum Zn concentration was, however, lower in the rabbits fed the Zn-deficient diet (approximately .35 micrograms/ml compared with .8 micrograms/ml for rabbits fed the low-Zn diet and 1.4 micrograms/ml for rabbits fed the control diet) . Tissue Zn concentrations generally declined in rabbits fed the low- and Zn-deficient diets, but this response depended on the particular tissue and diet . Zinc levels in bone decreased by approximately 45% and in fur by 20 to 30% on either low-Zn or Zn-deficient treatments . With a Zn-deficient diet, Zn in liver and testes decreased by 20%, Zn in skin by 35%, and Zn in brain by 10% . The Zn concentration in the skeletal muscle and thymus was, however, maintained . In the eye, Zn concentration in the aqueous humor declined by approximately 20% in rabbits fed the Zn-deficient diet.(ABSTRACT TRUNCATED AT 250 WORDS)

Zentralbl Veterinarmed B, 1991 Dec, 38(10), 721 - 30
In vitro binding of Pasteurella multocida cell wall preparations to tracheal mucus of cattle and swine and to a tracheal epithel cell wall preparation of cattle; Botcher L et al.; Outer membrane preparations of various Pasteurella isolates (Pasteurella multocida and some other Pasteurella species) from cattle and swine were extracted by N-lauryl-sarcosine sodium salt . Capsular extracts were prepared by heat treatment . Both preparations bound to epithel cell wall preparations (ECW) of trachea from cattle and to tracheal mucus of cattle and swine . Binding was demonstrated by an enzyme-linked immunosorbent assay (ELISA) . Distinct high adherence values were shown by the greater part of membrane preparations of mucoid Pasteurella strains, especially when originating from cattle.

J Rheumatol, 1991 Dec, 18(12), 1890 - 2
Report of 4 cases of Pasteurella multocida septic arthritis; Chevalier X et al.; Pasteurella multocida is frequently responsible for infections in man due to wounds inflicted by animals (generally cats or dogs) . However, the development of septic arthritis is a rare complication . We report 4 cases of Pasteurella multocida septic arthritis with demonstration of the organism in the joint in each case . Two cases presented with monoarthritis (sternoclavicular joint and wrist) and 2 cases had polyarticular involvement from the outset.

Am J Vet Res, 1991 Dec, 52(12), 2016 - 22
Characterization and comparison of antimicrobial susceptibilities and outer membrane protein and plasmid DNA profiles of Pasteurella haemolytica and certain other members of the genus Pasteurella; Rossmanith SE et al.; The outer membrane protein (OMP), plasmid, and antimicrobial resistance profiles of Pasteurella haemolytica serotypes 1 through 12, a bovine isolate of P multocida, a chicken isolate of P multocida, and an unidentified Pasteurella species of bovine origin were examined . Isolates of P haemolytica serotypes belonging to the same biotype possessed similar OMP profiles . Biotype A isolates contained 2 prominent OMP of 43 kilodaltons (kD) and 29 kD, whereas biotype-T serotypes contained 3 major OMP of 43, 36, and 25 kD . The major OMP profiles of the 2 P multocida isolates and the unidentified Pasteurella species were different from each other and from P haemolytica isolates . Plasmid DNA screening indicated both plasmid-containing and plasmid-free P haemolytica and P multocida isolates . Multiple drug resistance was found in pasteurellae isolates with and without plasmids . However, a relationship between drug resistance and plasmid isolation was found in 3 of 4 haemolytica serotype 1 field isolates, all of which contained a 2.51-megadalton plasmid and had multiple drug resistance for benzylpenicillin, ampicillin, streptomycin, and tetracycline.

Lab Anim Sci, 1991 Dec, 41(6), 572 - 6
Efficacy of enrofloxacin in the treatment of respiratory pasteurellosis in rabbits; Broome RL et al.; Fourteen New Zealand White rabbits with respiratory signs of naturally occurring Pasteurella multocida infections were treated with either an injectable or water-soluble oral formulation of enrofloxacin . Antimicrobial efficacy was evaluated by scoring clinical signs in the rabbits and recovery of the organisms . Seven (87%) of eight rabbits treated with the injectable regimen (5 mg/kg every 12 hours for 14 days) became culture-negative with no clinical signs within 72 hours after initiation of treatment . Six rabbits treated with the oral formulation (200 mg/liter of drinking water for 14 days) also became culture-negative and had no clinical signs 3 to 7 days after treatment began, but P . multocida was recovered from several sites in three (50%) rabbits . No rabbits showed any signs of gastroenteric disturbances.

Infect Immun, 1991 Nov, 59(11), 4212 - 20
Identification of RTX toxin target cell specificity domains by use of hybrid genes; Forestier C et al.; The Escherichia coli hemolysin (HlyA) and Pasteurella haemolytica leukotoxin (LktA) are cytolytic toxins encoded by genes belonging to the recently described RTX gene family . These cytotoxins are, respectively, 1,023 and 953 amino acids in length and are encoded by genes within identically organized operons . They share 45% amino acid sequence identities but differ in their target cell specificities . In vitro-derived recombinant hybrid genes between hlyA and lktA were constructed by using restriction endonuclease sites created by oligonucleotide site-directed mutagenesis . The cytolytic activity of hybrid proteins was investigated using as targets sheep erythrocytes and two cultured cell lines from different species (BL3, bovine leukemia-derived B lymphocytes; and Raji, human B-cell lymphoma cells) . HlyA is cytolytic to all three cell types . LktA lyses only BL3 cells . Among the hybrid proteins displaying cytolytic activity, the striking finding is that the hemolytic activity of several LktA-HlyA hybrids was independent of any cytolytic activity against either cultured cell species . The hemolytic activity was associated with the HlyA region between amino acids 564 and 739 . Structures that are critical for HlyA cytolytic activity against BL3 or Raji cells were destroyed when LktA-HlyA and HlyA-LktA hybrids were made, respectively, at amino acid positions 564 and 739 of HlyA . In contrast to HlyA, which lysed the two different cultured cell lines with equal efficiency, Lkt-HlyA hybrids possessing the amino-terminal 169 residues of LktA lysed BL3 cells more efficiently than Raji cells . This suggests that a significant but not exclusive element of the LktA ruminant cell specificity resides in the amino-terminal one-fifth of the protein . A molecular model of the functional domains of HlyA and LktA is presented.

J Cell Biol, 1991 Nov, 115(4), 949 - 58
A novel approach to detect toxin-catalyzed ADP-ribosylation in intact cells: its use to study the action of Pasteurella multocida toxin; Staddon JM et al.; Certain microbial toxins are ADP-ribosyltransferases, acting on specific substrate proteins . Although these toxins have been of great utility in studies of cellular regulatory processes, a simple procedure to directly study toxin-catalyzed ADP-ribosylation in intact cells has not been described . Our approach was to use {2-3H}adenine to metabolically label the cellular NAD+ pool . Labeled proteins were then denatured with SDS, resolved by PAGE, and detected by flurography . In this manner, we show that pertussis toxin, after a dose-dependent lag period, {3H}-labeled a 40-kD protein intact cells . Furthermore, incubation of the gel with trichloroacetic acid at 95 degrees C before fluorography caused the release of label from bands other than the pertussis toxin substrate, thus, allowing its selective visualization . The modification of the 40-kD protein was ascribed to ADP-ribosylation of a cysteine residue on the basis of inhibition of labeling by nicotinamide and the release of {3H}ADP-ribose from the labeled protein by mercuric acetate . Cholera toxin catalyzed the {3H}-labeling of a 46-kD protein in the {2-3H}adenine-labeled cells . Pretreatment of the cells with pertussis toxin before the labeling of NAD+ with {2-3H}adenine blocked {2-3H}ADP-ribosylation catalyzed by pertussis toxin, but not that by cholera toxin . Thus, labeling with {2-3H}adenine permits the study of toxin-catalyzed ADP-ribosylation in intact cells . Pasteurella multocida toxin has recently been described as a novel and potent mitogen for Swiss 3T3 cell and acts to stimulate the phospholipase C-mediated hydrolysis of polyphosphoinositides . The basis of the action of the toxin is not known . Using the methodology described here, P . multocida toxin was not found to act by ADP-ribosylation.

Microb Pathog, 1991 Nov, 11(5), 373 - 8
In vivo expression of iron regulated outer-membrane proteins in Pasteurella haemolytica-A1; Morck DW et al.; Pasteurella haemolytica-A1 was grown in vitro under iron-rich conditions, iron-depleted conditions, and in vivo within a chamber implanted in the peritoneal cavity of a rabbit to determine if iron regulated outer-membrane proteins were expressed in vivo . The antigenicity of outer membrane (OM) proteins from bacteria grown under these conditions was assessed by immunoblotting with pooled serum from convalescent bovine calves experimentally infected with P . haemolytica-A1 and serum from the implanted rabbit . Pasteurella haemolytica-A1 grown under iron-depleted conditions showed three distinct OM protein bands (71, 77, and 100 kDa) that were present in much lesser amounts when the organism was grown under iron-rich conditions . These same three bands were evident in OM protein preparations from bacteria grown in vivo . Western blotting indicated that these protein bands were recognized immunologically by the convalescent bovine serum and by serum from the implanted rabbit, in cells grown under the iron-depleted conditions and in vivo, but not if the bacteria were grown under the in vitro iron-rich conditions.

Antimicrob Agents Chemother, 1991 Nov, 35(11), 2419 - 22
Plasmid-mediated ROB-1 beta-lactamase in Pasteurella multocida from a human specimen; Rosenau A et al.; A Pasteurella multocida human isolate was resistant to beta-lactams because of production of ROB-1 beta-lactamase . The beta-lactamase was encoded by a 4.3-kb plasmid closely related to that of a Pasteurella bovine strain, as shown by Sau3A restriction profile and hybridization with a plasmid probe containing the blaROB-1 gene.

Am J Vet Res, 1991 Nov, 52(11), 1842 - 7
Market stress-associated changes in serum complement activity in feeder calves; Purdy CW et al.; Classical hemolytic complement (C) of calves was analyzed during a protocol designed to imitate the usual market handling of feeder calves from the southeastern United States . Serum C concentrations of the calves (n = 100 x 4 years) were evaluated on their farm of origin, on arrival at an auction market, on arrival at a feedyard, and during their first 4 weeks in the feedyard . Complement concentrations (measured in CH50 units) were typically lowest at the farm of origin and highest when the calves entered the auction market 28 to 133 days later . Serum C concentrations decreased after the calves encountered the severe stresses of being in the auction market for 7 days, 24-hour truck transport (1,932 km) to the feedyard, and the first 7 days in the feedyard . The C concentrations recovered after 21 to 28 days in the feedyard . Steers had significantly (P less than or equal to 0.05) lower C concentrations than did heifers in 3 of 4 years at the farm of origin, and in 2 of 4 years at the auction market . Morbid calves had significantly (P less than or equal to 0.05) lower C values than did healthy calves on day 7 in the feedyard in 3 of 4 years . There were significant differences in C concentrations of calves from different farms of origin in each of the 4 years . There was no significant difference in C concentrations of calves that were vaccinated vs those not vaccinated with Pasteurella haemolytica.

Am J Vet Res, 1991 Nov, 52(11), 1774 - 8
Effect of Pasteurella haemolytica saline capsular extract on bovine pulmonary endothelial cells; Kumar S et al.; The purpose of this in vitro study was to determine whether Pasteurella haemolytica capsular extract (CE) damages bovine pulmonary endothelial cells (EC) directly or through neutrophil-mediated mechanisms . Chromium 51-labeled EC were treated with the following variables: CE (1, 10, and 100 ng of protein/ml), CE and bovine neutrophils (10(6) cells/well), and CE and polymyxin B (500 U/ml) . Although only minimal damage to EC occurred by 5 hours after treatment, by 22 hours after treatment, the 10-ng and 100-ng CE dose produced severe damage to EC, as indicated by 51Cr release, cellular detachment, and loss of monolayer confluency . The component in the CE that was toxic to the EC was lipopolysaccharide, evidenced by effective neutralization of the toxic effect with polymyxin B . Neutrophils inhibited the CE-mediated EC toxicity and were activated, as indicated by shape change and adhesion to EC monolayers . We concluded that the lipopolysaccharide component of CE causes direct damage to EC, which can be attenuated by neutrophils and polymyxin B.

J Gen Microbiol, 1991 Nov, 137 ( Pt 11), 2663 - 8
Effects of antibiotics on the growth and morphology of Pasteurella multocida; Jacques M et al.; The effects of subminimal inhibitory concentrations (subMICs) of certain antibiotics, namely penicillin G, tetracycline and trimethoprim/sulphamethoxazole, on the growth and morphology of Pasteurella multocida were evaluated . SubMICs of penicillin markedly reduced the growth of P . multocida . Tetracycline and trimethoprim/sulphamethoxazole had no effect on its growth . SubMICs of penicillin greatly affected the morphology of P . multocida . At the highest concentrations tested (1/2 and 1/4 MIC) cells were acapsulate, and long filamentous cells (4-6 microns) were observed with some isolates . There was no correlation between the observed differences in the penicillin-binding proteins of the P . multocida isolates, and the extent of cell filamentation induced by penicillin G . SubMICs of tetracycline and trimethoprim/sulphamethoxazole did not seem to affect capsule production although filamentation was observed . Our results indicate that subMICs of penicillin can reduce growth of P . multocida . Furthermore, results also indicate that subMICs of antibiotics can affect the production of capsular material and the morphology of P . multocida.

Res Vet Sci, 1991 Nov, 51(3), 254 - 7
A quantitative measurement of the effect of avian influenza virus on the ability of turkeys to eliminate Pasteurella multocida from the respiratory tract; Sivanandan V et al.; The effect of avian influenza virus (AIV) infection on the ability of turkeys to eliminate Pasteurella multocida from the respiratory tract was evaluated . Four-week-old turkeys were experimentally infected with an apathogenic AIV subtype (H5N2) by the oculonasal route and subsequently superinfected with P multocida (Urbach strain) by the intranasal route three days after infection with AIV . Quantitative clearance of P multocida from the trachea and lung was determined using a pour plate technique on samples collected at intervals after infection . Samples from turkeys which had been infected with AIV were found to yield more P multocida than those from turkeys which had not been infected with AIV . The numbers of P multocida increased in infected birds to a greater extent than in birds which had not been infected with the virus . The present study suggests that AIV infection may contribute to the increased numbers and a decreased clearance of P multocida in turkeys.

Br Vet J, 1991 Nov-Dec, 147(6), 565 - 8
Experimental infection of dexamethasone-treated goats with Pasteurella haemolytica A2; Zamri-Saad M et al.; Sixteen goats either subjected to transport stress or without transport stress were treated with dexamethasone for 3 days prior to infection with P . haemolytica serotype A2 intranasally . The transport-stressed and dexamethasone-treated goats in the first group had various degrees of pulmonary lesions and the organism was re-isolated from the nasal cavity, lymph nodes and lungs . None of the goats treated with dexamethasone only were infected with P . haemolytica and had no lesions of pneumonic pasteurellosis . Treatment with dexamethasone alone failed to induce experimental infection by P . haemolytica except in combination with another stress factor.

Vet Microbiol, 1991 Nov, 29(3-4), 267 - 80
Characterization of the immunogenicity of formaldehyde detoxified Pasteurella multocida toxin; Bording A et al.; The immunogenicity of the Pasteurella multocida toxin (PMT) was studied in murine model systems . Mice were vaccinated with either formaldehyde treated pure PMT (pure toxoid) or formaldehyde treated crude extract of toxigenic P . multocida (crude toxoid) . The corresponding mean anti-PMT titres, sero-conversion rates and survival rates after challenge with affinity purified PMT were compared . When assessed both by anti-PMT titres and seroconversion and challenge, pure toxoid was a more potent immunogen than crude toxoid . This greater immunogenic potency was unaffected by the addition of killed cell preparations of Bordetella bronchiseptica, non-toxigenic P . multocida and B . pertussis . Increasing anti-PMT titres and seroconversion rates were induced by increasing doses of formaldehyde treated PMT (fPMT) in the pure toxoid vaccines, but not in the vaccines containing crude toxoid . However, improved survival rates were observed for both types of vaccine, when the fPMT content was raised . Immunization of pregnant mice with vaccines containing fPMT induced protection of the offspring against challenge with PMT; the protection of the offspring corresponded to that of the mother.

J Comp Pathol, 1991 Nov, 105(4), 455 - 65
The experimental production of mastitis in sheep by intramammary inoculation of Pasteurella haemolytica; el-Masannat ET et al.; Acute mastitis was consistently produced in primiparous ewes by inoculation of the mammary gland, via the teat canal, with an isolate of Pasteurella haemolytica, serotype A9, originating from a field case of ovine mastitis . Mastitis developed following the inoculation of as few as 10 colony forming units of this isolate, suggesting that only a small number of organisms would be required to initiate the disease under natural conditions provided they were already beyond the teat canal . Clinical signs and macroscopic lesions were well developed within 24 h of inoculation and were similar to those found in the naturally-occurring disease . The ability to reproduce mastitis consistently will facilitate studies of the pathogenesis of the disease and the comparison of different isolates of P . haemolytica with respect to virulence determinants.

Vet Clin North Am Food Anim Pract, 1991 Nov, 7(3), 669 - 94
Therapy of bovine bacterial pneumonia; Clarke CR et al.; Practical strategies for developing rational therapeutic regimens based on in vitro sensitivity and pharmacokinetic disposition are presented . Special attention is given to Pasteurella haemolytica, which is regarded as the most frequent cause of bovine bacterial pneumonia . Bacterial-dependent and host-dependent causes of therapeutic failure and potentially valuable novel therapies and drug combinations are considered.

Poult Sci, 1991 Nov, 70(11), 2259 - 66
The fate of Pasteurella multocida after intratracheal inoculation into turkeys; Matsumoto M et al.; Young adult turkeys were intratracheally inoculated with the P-1059 strain of Pasteurella multocida, and the fate of the organism was studied by quantifying the organism in various respiratory and systemic tissues at various times after the inoculation . The results showed that, in the first 2 h, on the order of 10(8) to 10(9) organisms deposited at the upper trachea multiplied in situ to gradually spread downwards to the lower respiratory tract . In the majority of turkeys, by 6 h postinoculation the organism invaded the circulatory system and multiplied vigorously in the liver and spleen . In some birds, however, the organism appeared to have reached the liver from the trachea instantaneously by an unidentified mechanism . Vaccination with inactivated vaccines protected turkeys from intratracheal challenge exposure, as well as from intramuscular inoculation.

Vaccine, 1991 Nov, 9(11), 817 - 24
Detection of stable epitopes on formaldehyde-detoxified Pasteurella multocida toxin by monoclonal antibodies; Foged NT; Progressive atrophic rhinitis in pigs can be prevented by vaccination of pregnant sows with formaldehyde-detoxified preparations of either crude extract of toxigenic Pasteurella multocida or purified P . multocida toxin (PMT) . The protective value of a vaccine is expected to be related to its content of detoxified immunogenic PMT, but previously described methods for detection of PMT cannot be used for the quantification of formaldehyde-detoxified PMT . In contrast to the epitopes on PMT recognized by previously produced anti-PMT monoclonal antibodies (mAbs), two of the epitopes recognized by mAbs developed in this study showed a significant stability after treatment with formaldehyde under conditions relevant for production of vaccines . When analysed in a sandwich ELISA based on these two mAbs, the titre of a preparation of PMT, detoxified by treatment with 1% (w/v) formaldehyde for 48 h at 20 degrees C, decreased by less than 30% when compared to the titre of native PMT . A close relationship between the amount of formaldehyde-treated PMT in a vaccine determined by the sandwich ELISA and its immunogenic properties in mice was observed.

Poult Sci, 1991 Oct, 70(10), 2023 - 7
Estimates of quantitative genetic parameters of immunological traits in the chicken; Cheng S et al.; Three in vivo assays were used to measure the immunocompetence of chickens in two generations of a selection experiment . The obtained data were used to estimate the variance components for sire and dam for antibody production to Pasteurella multocida and Mycoplasma gallisepticum vaccines, for T-cell-mediated immunity evaluated by a phytohemagglutinin wing web assay, and for clearance of foreign particles from the circulatory system . Heritabilities of and genetic correlations among these immunological traits were calculated from the sire variance components . Heritability estimates of the immunological traits based on the sire component of variance ranged from .06 to .53, and genetic correlations among immunological traits were generally negative.

Infect Immun, 1991 Oct, 59(10), 3626 - 9
Experimental model of atrophic rhinitis in gnotobiotic pigs; Ackermann MR et al.; To study the pathogenesis of atrophic rhinitis, gnotobiotic pigs (n = 6) were inoculated intranasally with a sterile sonicate of a toxigenic strain of Bordetella bronchiseptica (0.16 mg of protein per ml) at 5 days of age, and they were then inoculated intranasally with 1 ml (5,250 CFU/ml) of a live, toxigenic strain of Pasteurella multocida at 7 days of age . Pigs were necropsied at 2, 5, 9, 14, 21, and 28 days postinoculation; those pigs necropsied after 5 days had developed turbinate atrophy . Other gnotobiotic pigs received the following inoculation protocols: (i) a sterile sonicate of a nontoxigenic strain of B . bronchiseptica (0.2 mg of protein per ml), followed by toxigenic P . multocida (n = 4); (ii) toxigenic P . multocida alone (n = 7); (iii) diluent (sterile tryptose broth) (n = 2); (iv) the sterile sonicate of toxigenic B . bronchiseptica alone (n = 2); or (v) the sterile sonicate of a nontoxigenic strain of B . bronchiseptica alone (n = 2) . Turbinate atrophy did not occur in the latter groups except for one pig inoculated with only toxigenic P . multocida . These studies show that turbinate atrophy occurs in pigs given the toxigenic B . bronchiseptica sonicate and then given live, toxigenic P . multocida . This experimental regimen is a useful model for (i) studying the pathogenesis of atrophic rhinitis and (ii) testing vaccine strategies.

Infect Immun, 1991 Oct, 59(10), 3398 - 406
Molecular studies of Ssa1, a serotype-specific antigen of Pasteurella haemolytica A1; Lo RY et al.; A serotype-specific antigen of Pasteurella haemolytica A1 encoded on the recombinant plasmid pSSA1 is characterized . Nucleotide sequence analysis of the insert DNA in pSSA1 identified the gene ssaI, which codes for a protein of approximately 100 kDa . In vivo labeling of pSSA1-encoded protein in Escherichia coli maxicells showed the expression of a 100-kDa protein from the insert DNA on the recombinant plasmid . Northern blot and primer extension analyses were used to identify the mRNA transcript in P . haemolytica A1 and the putative promoter of ssaI . The antigen (designated Ssa1) could be localized to the outer membrane of P . haemolytica A1 and E . coli clones carrying pSSA1 . A rabbit serum against Ssa1 was produced by using whole cells of E . coli expressing Ssa1 on the surface as the immunogen, demonstrating that Ssa1 is immunogenic in rabbits . The results from colony immunoblot analysis with calf serum from animals that were resistant to P . haemolytica A1-induced pneumonia suggest indirectly that Ssa1 is also immunogenic in the animals.

Can J Vet Res, 1991 Oct, 55(4), 377 - 9
Effects of purified Pasteurella multocida dermonecrotoxin on the nasal ventral turbinates of fattening pigs: histological observations; Martineau-Doize B et al.; Fattening specific pathogen-free derived pigs were injected intramuscularly with dermonecrotoxin of Pasteurella multocida, capsular type D . Ten days later, the nasal ventral turbinates and liver were examined histologically . A moderate turbinate atrophy was observed due to an increased number of osteoclasts and the absence of intramembranous bone apposition . Liver lesions were limited to some hepatocyte necrosis, sinusoid neutrophil infiltration and Kupffer cell hypertrophy . This study demonstrated that adult pigs are sensitive to P . multocida dermonecrotoxin.

Am J Vet Res, 1991 Oct, 52(10), 1684 - 7
Growth hormone concentrations in plasma of healthy pigs and pigs with atrophic rhinitis; Ghoshal NG et al.; Plasma concentrations of porcine growth hormone (PGH) were similar in healthy pigs and those with atrophic rhinitis (AR), therefore, observed reduced growth rates and feed efficiency in naturally infected pigs with AR were not attributed to low concentrations of plasma PGH . Also, pituitary glands in both groups of pigs were responsive to growth hormone-releasing hormone (GHRH) challenge by increasing PGH secretion . Administration of clonidine hydrochloride to pigs naturally infected with AR failed to elicit any significant change (5.3 +/- 1.4 ng/ml) in the plasma concentration of PGH within a 45-minute bleeding interval . The pretreatment concentrations of PGH were similar in specific-pathogen-free toxin-treated and specific-pathogen-free control groups, but they increased significantly in toxin-treated pigs (20.7 +/- 8.2 ng/ml) within 15 minutes after GHRH injection . Porcine growth hormone release in toxin-treated pigs was variable; however, all pigs did not respond to GHRH administration: 3 responded with an increase in PGH release (35.6 +/- 10.6 ng/ml), 2 did not respond (6.7 +/- 0.5 ng/ml), and 1 had a decrease in PGH release (3.9 ng/ml) . Therefore, the observed reduced growth rates reported in the literature may be attributed to factors at the target level of PGH action, such as insufficient or down-regulation of PGH receptors, changes or impaired ability in the PGH receptor-binding characteristics, and inability of PGH receptor complex to transduce signal . Toxins are known to modulate signal transduction pathways . It has been speculated that serotype-D Pasteurella multocida toxin may influence growth by its effect on signal transduction from PGH receptor complex on the cell membrane to the interior of the cell.(ABSTRACT TRUNCATED AT 250 WORDS)

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