J Vet Med Sci, 2000 Jan, 62(1), 49 - 52 Decreased apolipoprotein C-III concentration in the high-density lipoprotein fraction from calves inoculated with Pasteurella haemolytica and bovine herpes virus-1; Yamamoto M et al.; Lipoprotein lipid and apoprotein concentrations are known to be altered during the acute-phase response . We have previously shown that the serum activity of lecithin:cholesterol acyltransferase (LCAT) and concentration of cholesteryl esters, both constituents of the high-density lipoprotein (HDL) fraction, are reduced in calves inoculated with Pasteurella haemolytica and bovine herpes virus-1, the two major pathogens for calf pneumonia . The concentration of apolipoprotein C-III (apoC-III), a low molecular mass protein component distributed mainly in the HDL fraction, was therefore examined in bacteria- and virus-inoculated calves . An enzyme-linked immunosorbent assay demonstrated that it was decreased by inoculations of Pasteurella haemolytica and bovine herpes virus-1 . The decrease was detected as early as 1 day after inoculation in both groups . A decreased serum apoC-III concentration was also observed by immunoblot analysis . It was detected in the HDL fractions from the bacteria- and virus-inoculated calves, and HDL apoC-III concentrations in the inoculated calves were decreased compared with controls . These results, coupled with the previous findings on LCAT activity and the cholesteryl ester concentration, indicate that a decreased HDL concentration is one of the early events occurring during the acute-phase response evoked by infections with Pasteurella haemolytica and bovine herpes virus-1. J Vet Med Sci, 2000 Jan, 62(1), 37 - 41 Detection of annexins I and IV in bronchoalveolar lavage fluids from calves inoculated with bovine herpes virus-1; Katoh N; Annexins are phospholipid-binding proteins and are abundant in the lung . Annexins I and IV, but not II and VI, have been detected in bronchoalveolar lavage (BAL) fluids from calves inoculated with Pasteurella haemolytica, the pathogen for calf pneumonia . In this study, BAL fluids from calves with experimental pneumonia induced by inoculation to right lung lobes of bovine herpes virus-1 (BHV-1), the major viral pathogen for pneumonia, were examined for detection of annexins I and IV . Of 6 calves inoculated with BHV-1, annexins I and IV were coincidentally detected in BAL fluids from right lung lobes of 4 calves, but not in BAL fluids from left lung lobes of 6 inoculated calves or those from left and right lung lobes of 3 control calves . Annexin II and VI were not found in any BAL fluids examined . These results, together with previous findings on calves inoculated with Pasteurella haemolytica, suggest that the release of annexins I and IV onto the alveolar surface is an essential event occurring in response to pulmonary infections of BHIV-1 and Pasteurella haemolytica. Vet Res Commun, 1999 Dec, 23(8), 467 - 73 The effects of dexamethasone on the response of bronchus-associated lymphoid tissue to intranasal administration of formalin-killed Pasteurella haemolytica A2 in goats; Zamri-Saad M et al.; A trial was conducted to observe the immediate and chronic effects in goats of dexamethasone administration on the bronchus-associated lymphoid tissue (BALT) response to intranasal administration of formalin-killed Pasteurella haemolytica A2 . Twenty-four goats were divided into four groups . Those in group 1 were injected intramuscularly with 1 mg/kg dexamethasone on three consecutive days, followed by intranasal exposure to formalin-killed P . haemolytica A2 one day after the last dexamethasone treatment . The goats in group 2 were similarly injected with dexamethasone followed by intranasal exposure to formalin-killed P . haemolytica A2 21 days after the last dexamethasone treatment . The animals in group 3 were exposed intranasally to formalin-killed P . haemolytica A2 without prior dexamethasone treatment . The animals in group 4 were untreated controls . The intranasal exposures to formalin-killed P . haemolytica A2 were repeated 2 weeks later . Intranasal exposure to formalin-killed P . haemolytica 1 day after dexamethasone treatment further reduced the number and size of BALT compared to the untreated control . Significantly (p < 0.01) more reduction of BALT occurred in goats exposed to formalin-killed P . haemolytica A2 21 days after dexamethasone treatment . On the other hand, intranasal exposure of goats without prior dexamethasone treatment stimulated the BALT compared to the untreated controls. Prev Vet Med, 2000 Jan 5, 43(1), 29 - 42 A stochastic partial-budget analysis of an experimental Pasteurella haemolytica feedlot vaccine trial; Gummow B et al.; A field trial compared a modified Pasteurella haemolytica biotype A serotype 1 leukotoxin vaccine to a commercial vaccine during March-July 1995 in a Natal Midlands, South African, feedlot . Weaners/long weaners purchased by the feedlot were allocated systematically into test vaccine and control vaccine groups of 1241 and 1240 head, respectively, and fed in groups of approximately 200 head . Morbidity and mortality were monitored until the animals were marketed . Details of pleuritis and pneumonia at veterinary meat inspection were recorded for 409 test-vaccinated and 424 control-vaccinated cattle . An increase in morbidity but not mortality risk of respiratory disease was shown between test (13.8% morbidity) and control (11.4% morbidity) groups . Cattle with a processing weight <245 kg were 1.4 times more likely to develop respiratory diseases than cattle with a processing weight > or =245 kg . Cattle bought on auction were 1.6 times more likely to develop respiratory disease than cattle bought at private sales . A partial farm budget incorporating Latin Hypercube sampling of uncertain variables was done to obtain the distribution of possible financial outcomes if the test vaccine were used . Impact (sensitivity) analyses indicated that median weight of carcass cut away had the greatest impact on the profit margin . The partial farm budget highlighted the importance of reducing sub-clinical lesions in a feedlot. J Antibiot (Tokyo), 1999 Nov, 52(11), 1007 - 16 In vitro microbiological characterization of novel cyclic homopentapeptides, CP-101,680 and CP-163,234, for animal health use; Norcia LJ et al.; Two cyclic homopentapeptides, CP-101,680 and CP-163,234 {6a-(3',4'-dichlorophenylamino) analogs of viomycin and capreomycin, respectively}, were identified as novel antibacterial agents for the treatment of animal disease, especially for livestock respiratory disease . The in vitro microbiological characterization of both CP-101,680 and CP-163,234 was carried out using their parent compounds, viomycin and capreomycin, as controls . This characterization included antibacterial spectrum, influence of media, inoculum size, pH, EDTA, polymixin B nonapeptide (PMBN), serum, cell-free protein synthesis inhibition, and time-kill kinetics . Our results indicated that the capreomycin analog, CP-163,234, showed slightly improved in vitro potency over the viomycin analog, CP-101,680 . Both analogs showed very potent cell-free protein synthesis inhibition activity and were bactericidal against Pasteurella haemolytica, P . multocida and Actinobacillus pleuropneumoniae at the level of 4 times and 8 times MICs . CP-163,234 was bactericidal at the level of 4x and 8x MIC against E . coli, but re-growth was observed after 24 hours incubation at both concentrations of CP-101,680. J Biol Chem, 2000 Jan 21, 275(3), 2239 - 45 Pasteurella multocida toxin stimulates mitogen-activated protein kinase via G(q/11)-dependent transactivation of the epidermal growth factor receptor; Seo B et al.; The dermatonecrotic toxin produced by Pasteurella multocida is one of the most potent mitogenic substances known for fibroblasts in vitro . Exposure to recombinant P . multocida toxin (rPMT) causes phospholipase C-mediated hydrolysis of inositol phospholipids, calcium mobilization, and activation of protein kinase C via a poorly characterized mechanism involving G(q/11) family heterotrimeric G proteins . To determine whether the regulation of G protein pathways contributes to the mitogenic effects of rPMT, we have examined the mechanism whereby rPMT stimulates the Erk mitogen-activated protein kinase cascade in cultured HEK-293 cells . Treatment with rPMT resulted in a dose and time-dependent increase in Erk 1/2 phosphorylation that paralleled its stimulation of inositol phospholipid hydrolysis . Both rPMT- and alpha-thrombin receptor- stimulated Erk phosphorylation were selectively blocked by cellular expression of two peptide inhibitors of G(q/11) signaling, the dominant negative mutant G protein-coupled receptor kinase, GRK2(K220R), and the Galpha(q) carboxyl-terminal peptide, Galpha(q)-(305-359) . Like alpha-thrombin receptor-mediated Erk activation, the effect of rPMT was insensitive to the protein kinase C inhibitor GF109203X, but was blocked by the epidermal growth factor receptor-specific tyrphostin, AG1478 and by dominant negative mutants of mSos1 and Ha-Ras . These data indicate that rPMT employs G(q/11) family heterotrimeric G proteins to induce Ras-dependent Erk activation via protein kinase C-independent "transactivation" of the epidermal growth factor receptor. Acta Med Okayama, 1999 Dec, 53(6), 245 - 52 Genomic structure of the rat major AP endonuclease gene (Apex) with an adjacent putative O-sialoglycoprotease gene (Prsmg1/Gcpl1) and a processed Apex pseudogene (Apexp1); Yao M et al.; Genomic sequencing and chromosomal assignment of the gene encoding rat APEX nuclease, a multifunctional DNA repair enzyme, were performed . An active Apex gene and a processed pseudogene were isolated from a rat genomic library . The active Apex gene consists of 5 exons and 4 introns spanning 2.1 kb . The putative promoter region of the Apex gene lacks the typical TATA box, but contains CAAT boxes and a CpG island having putative binding sites for several transcription factors, such as Sp1, AP-2, GATA-1 and ATF . A putative O-sialoglycoprotease (a homologue of Pasteurella haemolytica glycoprotease, gcp; abbreviated as Prsmg1/Gcpl1) gene consisting of 11 exons and 10 introns spanning 7.3 kb lies immediately adjacent to the Apex gene in a 5'-to-5' orientation . The Apex gene locus was mapped to rat chromosome 15p12 using in situ hybridization . The processed pseudogene (designated as rat Apexp1) has a nucleotide sequence 87.1% identical to that of the rat Apex cDNA, although several stop codons interrupting the coding sequences and multiple nucleotide deletions were observed . The Apexp1 is located in an inactive LINE sequence . Calculation of nucleotide substitution rates suggests that the immediate, active progenitor of Apexp1 arose 23 million years ago and that the non-functionalization occurred 15 million years ago. Am J Vet Res, 2000 Jan, 61(1), 51 - 6 Ultrastructural characterization of apoptosis in bovine lymphocytes exposed to Pasteurella haemolytica leukotoxin; Sun Y et al.; OBJECTIVE: To characterize ultrastructural changes of bovine lymphocytes exposed to Pasteurella haemolytica leukotoxin (LKT) . SAMPLE POPULATION: Partially purified LKT from a wild type P . haemolytica A1 strain and inactive pro-LKT from an isogeneic mutant Phaemolytica strain . Isolated bovine lymphocytes were obtained from 2 healthy calves . PROCEDURE: Isolated bovine lymphocytes were incubated with various concentrations of LKT and pro-LKT for 3 hours at 37 C and examined by use of transmission electron microscopy . A cytochemical Klenow DNA fragmentation assay was used to examine lymphocytes for DNA fragmentation . RESULTS: Lymphocytes incubated with LKT at a high concentration (1.0 toxic U/ml) had ultrastructural evidence of cytoplasmic and nuclear membrane rupture and swelling or lysis of mitochondria . Low concentrations of leukotoxin (0.1 toxic U/ml) induced DNA fragmentation in 80% of lymphocytes . Ultrastructurally, these cells had nuclear membrane blebbing, cytoplasmic vaculation, chromatin condensation, nuclear fragmentation, and membrane-bound apoptotic bodies . Incubation of lymphocytes with LKT at extremely low concentrations (0.001 toxic U/ml) or with pro-LKT did not alter their ultrastructure . Inclusion of 0.5 mM ZnCl2 in the medium blocked leukotoxin-induced ultrastructural changes in bovine lymphocytes . CONCLUSIONS AND CLINICAL RELEVANCE: Low concentrations of LKT induce apoptosis and high concentrations induce oncotic cell lysis in bovine lymphocytes . The ability of low LKT concentrations to induce apoptosis in host leukocytes may allow bacteria to escape host immune surveillance and colonize the host. J Med Microbiol, 2000 Jan, 49(1), 81 - 7 Restriction endonuclease analysis using Hhal and Hpall to discriminate among group B Pasteurella multocida associated with haemorrhagic septicaemia; Rimler RB; The purpose of this study was to improve and standardise restriction endonuclease analysis (REA) for discriminating isolates of serogroup B Pasteurella multocida associated with haemorrhagic septicaemia in wild and domestic animals and to create a reference database that can be used for epidemiological studies . Two techniques for extraction and isolation of chromosomal DNA were compared, a DNAzol method and an enzymic lysis followed by a two-phase partition method . No differences were observed between DNA fingerprint profiles with either technique; however, the former technique was faster and easier to perform . P . multocida isolated from different animals in different countries representing serotypes B:2, B:3, B:3,4 and B:4 were subjected to REA with HhaI and HpaII endonucleases . Forty-eight fingerprint profiles were distinguished among 222 isolates when only HhaI was used . By combining the data from REA with HhaI and HpaII used separately, 88 different groups could be distinguished among the same isolates . Following digestion with HhaI and electrophoresis, the DNA of all serotype B:2 isolates produced fingerprint profiles characterised by two trailing bands at approximately 8.4-7.1 kb which have not been observed in any other serotypes of P . multocida . Passage of three serotype B:2 isolates on laboratory media or two serotype B:2 isolates through mice did not result in a change of DNA fingerprint profile detectable by REA . The findings with 59 isolates from Sri Lanka showed that REA was highly discriminative in determining the genetic diversity of serotype B:2 P . multocida in an area where haemorrhagic septicaemia is endemic. Crit Care Med, 1999 Dec, 27(12), 2741 - 7 The risk of nosocomial pneumonia is not increased during partial liquid ventilation; Sajan I et al.; OBJECTIVE: To determine whether partial liquid ventilation (PLV) affects the risk of nosocomial pneumonia . STUDY DESIGN: To assess in vitro bacterial adhesion and viability after liquid perfluorocarbon exposure and to assess bacterial recovery after partial liquid ventilation in vivo in rabbits . SETTING: University animal research facility . SUBJECTS: Thirty-six New Zealand White rabbits . INTERVENTIONS: To assess adhesions, radiolabeled Escherichia coli were exposed to perfluorocarbon, incubated against artificial biosurfaces, and compared with nonexposed controls . Bacterial viability in vitro was assessed by exposing broth suspensions of Pasteurella multocida to perflubron for various times . Controls were run in parallel without exposure . Quantitative cultures were performed to determine viability . We undertook short-term and recovery in vivo investigations . The lungs of treated animals were filled with perflubron (approximately 18 mL/kg), and the control rabbits were ventilated without perflubron in an identical fashion . Cryopreserved aliquots of P . multocida were administered via an endotracheal tube . The short-term study animals were ventilated for 6 hrs before being killed . The recovery animals were ventilated for 2-4 hrs, extubated, and killed 20 hrs later . The lungs were removed, aseptically minced, and homogenized . Serial dilutions of the homogenate were quantitatively cultured by manual counting of colonies on agar plates . The recovered organisms were typed for species by the clinical microbiology laboratory . MEASUREMENTS AND MAIN RESULTS: The adhesion of bacteria to immobilized bronchoalveolar lavage and human saliva, respectively, was reduced by 65%+/-7% and 66%+/-1% (p < .05; n = 5) after exposure to perflubron and by 63%+/-9% and 68%+/-6% after exposure to FC-77 (p < .05; n = 5); however, adhesion was not affected by exposure to Rimar . There was no difference in bacterial viability between the control and perflubron-exposed bacteria (n = 5) . The in vivo study demonstrated a ten-fold or greater reduction in the number of recovered bacteria in the partial liquid ventilated group compared with the control group . CONCLUSIONS: This study suggests that different perfluorocarbons affect adhesions differently . Perflubron and FC-77 appear to decrease bacterial adhesion, whereas Rimar does not . Rerflubron does not have a direct bactericidal effect . Furthermore, PLV with perflubron decreased the number of viable bacteria per gram of tissue after an intentional inoculation of the airway, suggesting that the risk of nosocomial pneumonia is unlikely to be increased during PLV and may, in fact, be reduced in patients supported with PLV. J Clin Microbiol, 2000 Jan, 38(1), 327 - 32 Comparison of Pasteurella spp . simultaneously isolated from nasal and transtracheal swabs from cattle with clinical signs of bovine respiratory disease; DeRosa DC et al.; Twenty-four matched pairs of isolates of Pasteurella haemolytica and three matched pairs of isolates of Pasteurella multocida were isolated by using a nasal swab and a transtracheal swab from individual calves with clinical signs of bovine respiratory disease . The identity of each matched pair was confirmed biochemically and serologically . The similarity of the isolates obtained from a nasal swab and from a transtracheal swab was compared by using ribotyping and antibiotic susceptibility analyses . Although the calves were sampled only once with a nasal and a transtracheal swab, when both samples were bacteriologically positive the nasal swab identified the same bacterial species as the transtracheal swab 96% of the time . The nasal swab isolate was genetically identical to the transtracheal isolate in 70% of the matched pairs . Six different ribotypes were observed for the P . haemolytica isolates, while only one ribotype was observed for the limited number of P . multocida isolates . Of the six P . haemolytica ribotypes, two ribotypes predominated . All the paired isolates displayed similar susceptibility to ceftiofur, erythromycin, tilmicosin, trimethoprim-sulfamethoxazole, and florfenicol, with some minor variations for ampicillin and spectinomycin . These results suggest that a nasal swab culture can be predictive of the bacterial pathogen within the lung when the isolates are from an acutely ill animal and can be used to determine antibiotic susceptibility. Plasmid, 2000 Jan, 43(1), 99 - 102 Complete nucleotide sequence of a cryptic plasmid from the marine bacterium Vibrio splendidus and identification of open reading frames; Powers LG et al.; A 2.3-kb replication-proficient fragment was previously obtained from a cryptic plasmid (pPS41) isolated from a marine Vibrio splendidus isolate (P . A . Sobecky, T . J . Mincer, M . C . Chang, A . Toukdarian, and D . R . Helinski, 1998, Appl . Environ . Microbiol . 64, 2822-2830) . Analysis of the complete nucleotide sequence of plasmid pPS41 revealed two additional open reading frames (ORFs) . Analysis of ORF-1 revealed that its translated product has 125 amino acids with a predicted MW of 16,978 and ORF-2 encodes a putative protein of 151 amino acids with a predicted MW of 19,802 . The ORF-2 encoded protein showed 31 to 35% sequence homology to proteins identified to have a role in plasmid mobilization . These proteins are encoded on plasmids found in Escherichia coli and Pasteurella multocida . Plasmid pPS41 could be mobilized by a conjugative plasmid at frequencies of 1 x 10(-2) to 2 x 10(-2) . Acta Vet Scand, 1999, 40(3), 197 - 203 Effect of aerial ammonia on porcine infection of the respiratory tract with toxigenic Pasteurella multocida; Andreasen M et al.; The objective of the experimental study was to examine whether aerial ammonia alone could predispose the respiratory system of pigs to infection with toxigenic Pasteurella multocida type A . Two groups of 5 pigs each were continuously exposed to 50 ppm ammonia and less than 5 ppm ammonia, respectively, for a 59-day period (from 37 kg to 90 kg bodyweight) followed by necropsy . In an aerosol chamber all pigs were exposed to an aerosol of toxigenic P . multocida type A (mean bacterial concentration in the aerosol-exposure chamber: 10(5) colony forming units/m3; exposure period: 25 min) at day 10, 21, 35 and 49 after the onset of ammonia exposure . During the experiment none of the pigs showed clinical signs of pneumonia nor did they develop visible distortion of the snout . None of the pigs had gross lesions in the lungs at necropsy and toxigenic P . multocida was not detected by culture from the lungs from any of the pigs . The chance of recovering toxigenic P . multocida from nasal swabs (collected during experiment) was 2-4 times greater in the test group compared to the control group . The average daily weight gain was lower for the ammonia exposed pigs compared to the control group . In conclusion the results from this study suggest that ammonia in concentrations of 50 ppm is unlikely to predispose growing pigs to pulmonary infection with toxigenic P . multocida. Infect Immun, 2000 Jan, 68(1), 72 - 9 Lymphocyte function-associated antigen 1 is a receptor for Pasteurella haemolytica leukotoxin in bovine leukocytes; Jeyaseelan S et al.; Pasteurella (Mannheimia) haemolytica leukotoxin (Lkt) causes cell type- and species-specific effects in ruminant leukocytes . Recent studies indicate that P . haemolytica Lkt binds to bovine CD18, the common subunit of all beta2 integrins . We designed experiments with the following objectives: to identify which member of the beta2 integrins is a receptor for Lkt; to determine whether Lkt binding to the receptor is target cell (bovine leukocytes) specific; to define the relationships between Lkt binding to the receptor, calcium elevation, and cytolysis; and to determine whether a correlation exists between Lkt receptor expression and the magnitude of target cell cytolysis . We compared Lkt-induced cytolysis in neutrophils from control calves and from calves with bovine leukocyte adhesion deficiency (BLAD), because neutrophils from BLAD-homozygous calves exhibit reduced beta2 integrin expression . The results demonstrate for the first time that Lkt binds to bovine CD11a and CD18 (lymphocyte function-associated antigen 1 {LFA-1}) . The binding was abolished by anti-CD11a or anti-CD18 monoclonal antibody (MAb) . Lkt-induced calcium elevation in bovine alveolar macrophages (BAMs) was inhibited by anti-CD11a or anti-CD18 MAb (65 to 94% and 37 to 98%, respectively, at 5 and 50 Lkt units per ml; P < 0.05) . Lkt-induced cytolysis in neutrophils and BAMs was also inhibited by anti-CD11a or anti-CD18 MAb in a concentration-dependent manner . Lkt bound to porcine LFA-1 but did not induce calcium elevation or cytolysis . In neutrophils from BLAD calves, Lkt-induced cytolysis was decreased by 44% compared to that of neutrophils from control calves (P < 0.05) . These results indicate that LFA-1 is a Lkt receptor, Lkt binding to LFA-1 is not target cell specific, Lkt binding to bovine LFA-1 correlates with calcium elevation and cytolysis, and bovine LFA-1 expression correlates with the magnitude of Lkt-induced target cell cytolysis. Infect Immun, 2000 Jan, 68(1), 6 - 12 Characterization of PaxA and its operon: a cohemolytic RTX toxin determinant from pathogenic Pasteurella aerogenes; Kuhnert P et al.; Pasteurella aerogenes is known as a commensal bacterium or as an opportunistic pathogen, as well as a primary pathogen found to be involved in abortion cases of humans, swine, and other mammals . Using broad-range DNA probes for bacterial RTX toxin genes, we cloned and subsequently sequenced a new operon named paxCABD encoding the RTX toxin PaxA in P . aerogenes . The pax operon is organized analogous to the classical RTX operons containing the activator gene paxC upstream of the structural toxin gene paxA, which is followed by the secretion protein genes paxB and paxD . The highest sequence similarity of paxA with known RTX toxin genes is found with apxIIIA (82%) . PaxA is structurally similar to ApxIIIA and also shows functional analogy to ApxIIIA, since it shows cohemolytic activity with the sphingomyelinase of Staphylococcus aureus, known as the CAMP effect, but is devoid of direct hemolytic activity . In addition, it shows to some extent immunological cross-reactions with ApxIIIA . P . aerogenes isolated from various specimens showed that the pax operon was present in about one-third of the strains . All of the pax-positive strains were specifically related to swine abortion cases or septicemia of newborn piglets . These strains were also shown to produce the PaxA toxin as determined by the CAMP phenomenon, whereas none of the pax-negative strains did . This indicated that the PaxA toxin is involved in the pathogenic potential of P . aerogenes . The examined P . aerogenes isolates were phylogenetically analyzed by 16S rRNA gene (rrs) sequencing in order to confirm their species . Only a small heterogeneity (<0.5%) was observed between the rrs genes of the strains originating from geographically distant farms and isolated at different times. Trop Anim Health Prod, 1999 Dec, 31(6), 333 - 45 Studies on the respiratory disease 'sonbobe' in camels in the eastern lowlands of Ethiopia; Bekele T; New epidemics of respiratory disease have caused 29.6 morbidity and 6.4% mortality in camels in the Somalia region of Ethiopia . The major clinical signs observed were fever of 40-41.5 degrees C, depression, cough, loss of appetite and a watery nasal discharge that became mucopurulent at a later stage . Finally, the camel became recumbent and extended its neck straight along the ground . Some of the animals died within 8-9 days . The major post-mortem lesions were hydrothorax, adhesion of the lung to the thorax, red and grey hepatization, emphysema, hydropericardium and fibrinous pericarditis . A treatment trial indicated that oxytetracycline was more effective than a combination of penicillin and streptomycin, the results showing a significant difference (p < 0.05) between the treated and control groups . The bacteria isolated from lung, thoracic fluid and whole blood were Pasteurella haemolytica . Further studies on the epidemiology of this disease, the identification of the serotypes involved, and the demonstration of any primary viral initiating agent are recommended to allow the development of preventive methods. Am J Ophthalmol, 1999 Oct, 128(4), 514 - 5 Periocular abscess and cellulitis from Pasteurella multocida in a healthy child; Hutcheson KA et al.; PURPOSE: To examine an unusual cause of periorbital cellulitis, Pasteurella multocida . METHODS: Case report, review of the literature . RESULTS: We treated a 13-year-old previously healthy child who developed Pasteurella preseptal cellulitis secondary to a cat bite and cat scratch . After receiving a dose of intravenous antibiotics and starting oral antibiotics, the child had delayed onset of several abscesses around the right eye, with marked pain and erythema . After incision and drainage, he improved . CONCLUSION: Pasteurella multocida is a rare but potentially serious cause of ocular infection . All cases of potential exposure should be treated promptly and followed until complete resolution of infection. Glycobiology, 2000 Jan, 10(1), 31 - 7 Purification and characterization of an adhesin from Pasteurella haemolytica; Jaramillo L et al.; We purified an adhesin from Pasteurella . haemolytica by affinity chromatography using glutaraldehyde treated rabbit erythrocytes stroma . The adhesin is a protein of 68 kDa, as determined by SDS-PAGE, and the most abundant amino acids constituting this protein were Gly, Ser, Glx, and Ala, and low concentrations of Cys, Met, and Tyr residues were also found . The N-terminal sequence of the adhesin is ANEVNVYIYKQPYLI . No carbohydrate residues were detected . The adhesin agglutinated rabbit erythrocytes but when the latter were desialylated or pronase treated the agglutinating activity was abolished . The agglutinating activity of the adhesin was inhibited with N-acetyl-D-glucosamine (GlcNAc), and in a lesser degree with N-acetyl-neuraminic acid (NeuAc) . GalNAc, N-glycolyl-neuraminic acid, N-deacetylated GlcNAc, or neutral sugars do not modify the activity of the adhesin . The equatorial -OH on C4 and the NH-acetylated group on C2 from GlcNAc, as well as the 4-OH and NH-acetylated group on C5 from NeuAc seem to be responsible for the interaction with the adhesin . The protein is divalent cation-dependent and thermolabile . As for the agglutinating activity, the adhesion of P.haemolytica to tracheal cell-cultures was inhibited by GlcNAc, NeuAc or the purified adhesin, strongly suggesting that the P.haemolytica adhesin plays an important role in infection. Infect Immun, 1999 Dec, 67(12), 6264 - 9 Correlation of Pasteurella haemolytica leukotoxin binding with susceptibility to intoxication of lymphoid cells from various species; Sun Y et al.; Pasteurella haemolytica, the causative agent of shipping fever pneumonia in cattle, produces a leukotoxin (LKT) which lyses ruminant leukocytes with high efficiency but is reputed to not affect leukocytes from nonruminant species . In this study, we tested the supposition that LKT binding correlates positively with susceptibility to intoxication of susceptible isolated bovine lymphocytes and lymphoma tissue culture cells (BL3 cells) and negatively with reputed nonsusceptible equine, porcine, and canine lymphocytes and human lymphoid tissue culture cells (Raji cells) . Bovine lymphocytes and BL3 cells were highly susceptible to LKT intoxication, exhibiting both substantial increase in intracellular Ca(2+) concentration and marked leukolysis . Exposure of reputed LKT-nonsusceptible porcine lymphocytes and Raji cells to LKT caused a slightly increased intracellular Ca(2+) concentration but no leukolysis . No LKT effect was detected for equine and canine lymphocytes . LKT bound to lymphoid cells from all species tested . Intact 102-kDa LKT was recovered from exposed isolated lymphoid cell membranes . Pro-LKT acylation was not required for LKT binding to BL3 cells . LKT binding was rapid, with maximal binding occurring by 3 min, and was proportional to the LKT concentration in the range 0.04 to 4.0 microg/ml . For this LKT concentration range, BL3 cells bound more LKT than did porcine lymphocytes or Raji cells, suggesting that LKT binds to BL3 cells with higher affinity than to porcine lymphocytes or Raji cells . Above 4.0 microg/ml, LKT demonstrated saturable binding to BL3 cells . Neutralizing anti-LKT monoclonal antibody (MAb) MM601 diminished LKT binding to BL3 by 36% while decreasing leukolysis by 81% . In contrast, MM601 did not diminish LKT binding to Raji cells . Pretreatment of target cells with 120 microg of protease K per ml diminished LKT binding to BL3 cells by 75%, with only a 25% decrease in leukolysis . However, pretreatment with 150 microg of protease K per ml abolished the remaining 25% of LKT binding and 75% leukolysis . Therefore, P . haemolytica LKT binds rapidly to susceptible and to reputed nonsusceptible lymphoid cells . LKT binding resulting in species-specific leukolysis was characterized by high affinity, inhibition by MAb MM601, and relative resistance to protease K pretreatment of lymphoid cells . Two types of LKT binding to lymphoid cells are proposed . High-affinity binding leads to efficient leukolysis . In some lymphoid cells from reputed LKT-nonsusceptible species, low-affinity LKT binding may cause a low-efficiency increase in the intracellular Ca(2+) concentration without leading to leukolysis. Am J Vet Res, 1999 Nov, 60(11), 1402 - 6 Use of a polymerase chain reaction method to detect the leukotoxin gene lktA in biogroup and biovariant isolates of Pasteurella haemolytica and P trehalosi; Fisher MA et al.; OBJECTIVE: To determine whether Pasteurella haemolytica and P trehalosi isolates possess the structural gene for Pasteurella leukotoxin lktA and whether beta-hemolytic activity of these isolates correlated with detection of the lktA gene . SAMPLE POPULATION: 147 P haemolytica isolates from 21 biovariant groups and 101 P trehalosi isolates from 7 biovariant groups . In addition, P multocida and organisms from 7 other genera were tested to establish specificity of the procedure . PROCEDURE: Isolates were observed for beta-hemolysis . A polymerase chain reaction (PCR) procedure was used to amplify the RTX domain of the Pasteurella lktA gene . RESULTS: The lktA gene was detected in 108 (44%) isolates, including 15 associated with respiratory tract disease . All but 2 (98%) of the isolates that had the lktA gene were beta-hemolytic when grown on sheep blood agar . The remaining 140 isolates were negative for the lktA gene and hemolytic activity . CONCLUSIONS AND CLINICAL RELEVANCE: Hemolytic activity of P haemolytica and P trehalosi isolates correlated with detection of the lktA gene for all but 2 isolates . However, 56% of isolates tested were negative for the lktA gene and beta-hemolytic activity . Leukotoxin production and secretion is a major virulence factor when other conditions are favorable for disease development . Therefore, identification of strains that possess the lktA gene may aid in the evaluation of the pathogenic potential of Pasteurella strains carried by wild and domestic animals. Am J Vet Res, 1999 Nov, 60(11), 1390 - 5 Detection of annexin I and IV and haptoglobin in bronchoalveolar lavage fluid from calves experimentally inoculated with Pasteurella haemolytica; Katoh N et al.; OBJECTIVE: To determine whether annexins or haptoglobin could be detected in bronchoalveolar lavage (BAL) fluid specimens obtained from calves experimentally inoculated with Pasteurella haemolytica . ANIMALS: Twelve 2- to 3-month-old male Holstein calves . PROCEDURE: Pasteurella haemolytica was inoculated into the right lung lobes of each of 6 calves . Six other calves received vehicle alone and were used as control calves . Specimens of BAL fluid were obtained from 3 control and 3 inoculated calves 1 day after inoculation and from the other calves 2 days after inoculation . The amount of annexins I, II, IV, and VI, and haptoglobin in BAL fluid specimens was examined by use of immunoblot analysis . RESULTS: Annexins I and IV were detected in BAL fluid specimens obtained from the right lung lobes of each of the inoculated calves, but annexins II and VI were not . Annexin I also was found in BAL fluid specimens obtained from the left lung lobes of each inoculated calf and from left and right lung lobes of the control calves . By comparison, detection of annexin IV was essentially limited to the right lung lobes of inoculated calves . Haptoglobin was detected in some, but not all, BAL fluid specimens from the right lung lobes of inoculated calves, and its detection in BAL fluid was associated with serum proteins such as albumin . CONCLUSIONS AND CLINICAL RELEVANCE: Annexin IV was detected most specifically in response to inoculation of P haemolytica . This protein could be used as a marker for inflammatory pulmonary disease caused by P haemolytica. Int J Antimicrob Agents, 1999 Sep, 13(1), 47 - 51 Anionic antimicrobial peptide-lysozyme interactions in innate pulmonary immunity; Kalfa VC et al.; The respiratory tract contains numerous antimicrobial factors necessary for normal innate pulmonary defense . Although many of these molecules reside in airway surface liquid (ASL) simultaneously, little information exists concerning antagonistic, additive, or synergistic interactions . Since both cationic lysozyme and anionic antimicrobial peptides (AP) are found in high concentrations in ASL, the purpose of this study was to assess any interaction that might affect antimicrobial activity . For this, Pasteurella haemolytica, Micrococcus lysodeikticus, or Pseudomonas aeruginosa were added to egg white lysozyme (3.9-250.0 microg/ml) or human neutrophil lysozyme (0.8-50.0 microg/ml) and H-GADDDDD-OH (from 0.01 to 0.50 mM) mixtures in 50, 100, or 150 mM NaCl; incubated for 2 h; and then plated . In this assay, the MICs of AP for P . haemolytica, M . lysodeikticus, and P . aeruginosa varied slightly depending upon the concentration of NaCl and MICs generally increased slightly with increasing NaCl concentrations . The MIC of lysozyme for P . haemolytica and M . lysodeikticus also increased slightly with increasing NaCl concentrations . The MIC of lysozyme for P . aeruginosa was greater than 50 microg/ml and did not vary with increasing NaCl concentrations . When AP was combined with lysozyme in 50, 100, or 150 mM NaCl concentrations, there was no significant interaction that affected antimicrobial activity . In conclusion, the MICs of AP generally increased with increasing NaCl concentrations but lysozyme and AP appeared not to interact significantly at physiologically relevant concentrations. J Vet Med Sci, 1999 Oct, 61(10), 1101 - 6 Reduced serum lecithin:cholesterol acyltransferase activity and cholesteryl ester concentration in calves experimentally inoculated with Pasteurella haemolytica and bovine herpes virus-1; Nakagawa H et al.; Lecithin:cholesterol acyltransferase (LCAT), the enzyme responsible for esterification of cholesterol in plasma, is reported to be implicated in the regulation of inflammation in laboratory animals . The purpose of the present study was to elucidate the possible relevance of LCAT in the pathogenesis of calf pneumonia induced by inoculations of Pasteurella haemolytica and bovine herpes virus-1 into the calf lung . Serum LCAT activity was significantly (P < 0.01) reduced in calves inoculated with Pasteurella haemolytica . The concentration of cholesteryl esters (CE), the product of the LCAT reaction, was also decreased in the inoculated group . Decreases in LCAT activity and the CE concentration were similarly observed in calves in which bovine herpes virus-1 was inoculated . In both bacteria- and virus-inoculated calves, CE concentrations in the high-density lipoprotein fractions were distinctly decreased, whereas those in the low-density lipoprotein fractions were practically unaltered . The acute-phase proteins haptoglobin and serum amyloid A were detected in sera from the bacteria- and virus-inoculated calves; however, the two acute-phase proteins were also found in sera from the control calves . These results suggest that decreases in LCAT activity and the CE concentration are involved in the pathogenesis of pneumonia induced by inoculation of calves with Pasteurella haemolytica and bovine herpes virus-1, and also that the change in the LCAT system is more intimately related to the occurrence of calf pneumonia than the induction of acute-phase proteins such as haptoglobin. Med Clin (Barc), 1999 Oct 9, 113(11), 415 - 7 {Respiratory pasteurellosis . Description of a first series in Spain}; Ferrer A et al.; OBJECTIVE: To know the clinical and microbiological characteristics of a first series of 8 patients in whom Pasteurella multocida was detected in samples from low respiratory airways . PATIENTS AND METHODS: Patients with respiratory disease who had positive cultures for P . multocida in several biologic samples between 1986 and 1998 were studied . Patient's data were obtained from clinical files; microbiological study included microscopic examination, qualitative culture in all samples and quantitative culture in low respiratory airway samples . RESULTS: P . multocida was detected in 7 males and one female, with a mean age of 63 years (range: 6-71) . All but one had a previous bronchial disease: 5 had chronic obstructive bronchial disease, 2 had bronchiectasis and one had relapsing acute bronchitis . Three patients referred a previous contact with pets . The main diagnosis, at the time P . multocida was detected, was exacerbated bronchitis in 4 patients, pneumonia in 2 (one with sepsis and positive blood culture to P . multocida) and another one with empyema . In the only paediatric patient P . multocida was a casual finding . Two patients died, both having a severe immunosuppression . The percentage of P . multocida detection compared to all low respiratory airway isolates along the study period was 0.02% . All samples were purulent and more than 10(7) P . multocida colony-forming units/millilitre were detected . All strains were penicillin-sensitive . CONCLUSIONS: Pasteurella multocida respiratory infection is rare, although it is probably underestimated owing to the fact that it is difficult to identify when coexisting with oropharingeal flora . Characteristically, in affects old males with bronchial disease are involved and can be life threatening in severely immunosuppressed patients. Poult Sci, 1999 Nov, 78(11), 1532 - 5 Effect of selection for increased body weight on mitogenic responses in turkeys; Li Z et al.; Mitogenic responses were examined for purified peripheral blood mononuclear cells (PBMC) and whole blood from individuals in a line (F) of turkeys selected for increased 16-wk BW and its corresponding randombred control (RBC2) . The PBMC were isolated by centrifugation over Histopaque-1077 density gradient and tested for mitogenic responses to concanavalin A (Con A; 25 microg/mL) and phytohemagglutinin (PHA)-M; 100 microg/mL) . For the whole blood assay, 6-wk-old poults from both lines were injected with inactivated Pasteurella multocida . Heparinized blood samples were collected prior to injection (0 d) and at 2, 4, 7, and 14 d postinjection . The diluted whole blood was then tested for the mitogenic responses to Con A (25 microg/mL) and PHA-M (25 microg/mL) . The cultures were then pulsed with 3H-thymidine, and incorporation was measured using a liquid scintillation counter . There was a line difference in the mitogenic responses to Con A for PBMC and whole blood assays, but no line difference was observed in the response to PHA-M for both assays . For the purified PBMC assay, the F line had a lower response than its randombred control line (P < or = 0.05) to Con A expressed as either cpm or a stimulation index (SI; ratio of cpm for stimulated cells to the cpm for unstimulated cells) . For the whole blood assay, the F line had generally lower SI values in the responses to Con A than the RBC2 line, with differences being significant at 0 and 2 d postinjection (P < or = 0.01) and at 14 d postinjection (P < or = 0.05) . Genetic selection for increased BW might have affected the lymphoblastogenic potential of Line F that could affect disease resistance. Lab Anim Sci, 1999 Oct, 49(5), 551 - 9 Characterization of rabbit Pasteurella multocida isolates by use of whole-cell, outer-membrane, and polymerase chain reaction typing; Dabo SM et al.; PURPOSE: To characterize Pasteurella multocida isolates from laboratory rabbits using serotyping, sodium dodecyl sulfate-polyacrylamide gel electrophoresis of whole-cell proteins (WCPs) and outer-membrane proteins (OMPs), and polymerase chain reaction (PCR) fingerprinting . METHODS: Fifty isolates were obtained from five sources: ATCC (1), Oklahoma (4), Michigan (9), Minnesota (7), and Texas (29) . The PCR fingerprinting was conducted using two minisatellite probes for M13 and a modified M13 core sequence and two microsatellite probes--(GTG)5 and (GACA)4 . RESULTS: Forty-five isolates were serogroup A, and five were serogroup D . Ten WCP patterns (W1-W10) with one variation (W1a) and 10 OMP (OM1-OM10) patterns were found . Primers M13 phage, modified M13 phage, (GTG)5, and (GACA)4 generated 7, 9, 5, and 9 fingerprint types, respectively . Combination of WCP, OMP, and PCR fingerprint results yielded 39 groups with a discrimination index of 0.98 . The PCR fingerprint results generally indicated clonal association among isolates within geographic locations except for the isolates from Texas, which varied markedly in PCR fingerprint types . CONCLUSION: Single primer PCR fingerprinting provided a simple and rapid means of typing P . multocida isolates from laboratory rabbits . Combinations of conventional and molecular typing enhanced differentiation among P . multocida isolated from rabbits with pasteurellosis. APMIS, 1999 Oct, 107(10), 913 - 20 Typing of Pasteurella multocida from haemorrhagic septicaemia in Danish fallow deer (Dama dama); Aalbaek B et al.; Isolates of Pasteurella multocida ssp . multocida (n = 31) from a Danish population of fallow deer which succumbed to haemorrhagic septicaemia during 1992 1993 and isolates from the palatine tonsils of apparently healthy fallow deer from the same area (n=6) were typed and compared with P . multocida from other sources . Plasmids were net observed in the fallow deer strains and one unique pattern was observed by ribotyping using HindIII and by pulsed-field gel electrophoresis using SanlI as restriction endonuclease . All Danish fallow deer isolates belonged to serotype B:3,4 . On restriction endonuclease analysis using HhaI as restriction endonuclease, all had a profile identical to that of a fallow deer isolate from the United Kingdom: profile 0033 of Wilson et al . On restriction endonuclease analysis using HpaII as restriction endonuclease, the Danish fallow deer isolates had a unique profile, designated 0062, which differed slightly from that of a fallow deer isolate from the United Kingdom . P . multocida from other animal species were genotypically different from the fallow deer isolates . It is concluded that a specific clone of P . multocida was responsible for the outbreak of haemorrhagic septicaemia among Danish fallow deer . A carrier rate of 27% was demonstrated among apparently normal animals from the same population. FEMS Microbiol Lett, 1999 Nov 1, 180(1), 15 - 20 Reduced pH causes structural changes in the potent mitogenic toxin of Pasteurella multocida; Smyth MG et al.; Pasteurella multocida toxin is a potent mitogen that is believed to act intracellularly . On transverse urea gradient gels at pH 8.0 the toxin displayed one major unfolding transition at 4 M urea . However, at pH 6.1 the unfolding transition took place at 3.5 M urea . Circular dichroism spectra also indicated that a structural change took place at acidic pH . In addition it was found that the toxin that had been denatured in 8 M urea refolded in solution with a high recovery of biological activity . These findings are discussed in terms of the likely domain structure of the P . multocida toxin. Microb Pathog, 1999 Nov, 27(5), 311 - 27 Identification of in vivo induced genes in Actinobacillus pleuropneumoniae; Fuller TE et al.; We have developed an in vivo expression technology (IVET) system to identify Actinobacillus pleuropneumoniae gene promoters that are specifically induced in vivo during infection . This system is based upon an avirulent riboflavin-requiring A . pleuropneumoniae mutant and a promoter-trap vector (pTF86) that contains, in sequence, the T4 terminator, a unique Bam HI site, a promoterless copy of the V . harveyi luxAB genes, and a promoterless copy of the B . subtilis ribBAH genes in the E . coli - A . pleuropneumoniae shuttle vector pGZRS19 . Sau 3A fragments of A . pleuropneumoniae genomic DNA were cloned into the Bam HI site in pTF86 and transformed into the A . pleuropneumoniae Rib- mutant . Pigs were infected with pools of 300-600 transformants by endobronchial inoculation and surviving bacteria were isolated from the pigs' lungs at 12-16 h post-infection . Infection strongly selected for transformants containing cloned promoters which drove expression of the vector ribBAH genes and allowed survival of the Rib- mutant in vivo . Strains that survived in vivo, but which minimally expressed luciferase activity in vitro, should contain cloned promoters that are specifically induced in vivo . Ten clones, designated iviA-J, were isolated which contain promoters that are induced in vivo during infection . These ivi clones were shown to be induced in the animal by luminescence of infected tissue and by direct assay of bacteria recovered from bronchoalveolar lavage . Four of these clones were putatively identified by amino acid sequence similarity as ilvI, the ilvDA operon, the secE-nusG operon, and the mrp gene . This is the first report of an IVET system for use in the family Pasteurellaceae, as well as the first report of an IVET system utilizing an infection model of pneumonia in the natural host . J Bacteriol, 1999 Nov, 181(21), 6747 - 55 Bacterial phylogenetic clusters revealed by genome structure; Liu SL et al.; Current bacterial taxonomy is mostly based on phenotypic criteria, which may yield misleading interpretations in classification and identification . As a result, bacteria not closely related may be grouped together as a genus or species . For pathogenic bacteria, incorrect classification or misidentification could be disastrous . There is therefore an urgent need for appropriate methodologies to classify bacteria according to phylogeny and corresponding new approaches that permit their rapid and accurate identification . For this purpose, we have devised a strategy enabling us to resolve phylogenetic clusters of bacteria by comparing their genome structures . These structures were revealed by cleaving genomic DNA with the endonuclease I-CeuI, which cuts within the 23S ribosomal DNA (rDNA) sequences, and by mapping the resulting large DNA fragments with pulsed-field gel electrophoresis . We tested this experimental system on two representative bacterial genera: Salmonella and Pasteurella . Among Salmonella spp., I-CeuI mapping revealed virtually indistinguishable genome structures, demonstrating a high degree of structural conservation . Consistent with this, 16S rDNA sequences are also highly conserved among the Salmonella spp . In marked contrast, the Pasteurella strains have very different genome structures among and even within individual species . The divergence of Pasteurella was also reflected in 16S rDNA sequences and far exceeded that seen between Escherichia and Salmonella . Based on this diversity, the Pasteurella haemolytica strains we analyzed could be divided into 14 phylogenetic groups and the Pasteurella multocida strains could be divided into 9 groups . If criteria for defining bacterial species or genera similar to those used for Salmonella and Escherichia coli were applied, the striking phylogenetic diversity would allow bacteria in the currently recognized species of P . multocida and P . haemolytica to be divided into different species, genera, or even higher ranks . On the other hand, strains of Pasteurella ureae and Pasteurella pneumotropica are very similar to those of P . multocida in both genome structure and 16S rDNA sequence and should be regarded as strains within this species . We conclude that large-scale genome structure can be a sensitive indicator of phylogenetic relationships and that, therefore, I-CeuI-based genomic mapping is an efficient tool for probing the phylogenetic status of bacteria. Poult Sci, 1999 Oct, 78(10), 1377 - 9 Variation in resistance to Pasteurella multocida among turkey lines; Nestor KE et al.; Previous research has shown that a line (F) of turkeys selected long-term for increased 16-wk body weight was more susceptible to challenge with washed Pasteurella multocida than a randombred control line (RBC2), the base population of F . A previous study also indicated that the mortality of the F line following challenge with P . multocida was similar to that of sire lines from two of the three major primary breeders . The purpose of the present study was to compare the resistance of the sire line from the third major primary turkey breeder (C) not previously studied with that of the F and RBC2 lines to determine whether there is variation in resistance among the sire lines from three major primary breeders . The sire lines from all three primary breeders were used in the production of commercial turkeys . Body weights of the F line were greater than those of the C line at the time of challenge with P . multocida . Both the C and F lines were heavier than the RBC2 line . The birds were challenged at 6 wk of age with a field isolate of washed P . multocida (1.2 x 10(7) organism per bird of capsular serogroup A and somatic serotype 3,4) . Mortality was recorded daily for 14 d . Mortality following challenge with P . multocida was higher in the F line than in the C line, and both large-bodied lines had higher mortality than the RBC2 line . Based on the present results and those published in the literature, there may be variation in resistance among commercial sire lines from the three major primary breeders. Lancet, 1999 Oct 9, 354(9186), 1267 - 8 Beware of dogs licking ears; Godey B et al.; A patient with right-sided chronic purulent otorrhoea developed meningitis due to Pasteurella multocida transmitted by a dog that frequently licked his ear . We suggest that patients with a perforated tympanic membrane should avoid being licked on their ears by animals. Poult Sci, 1999 Sep, 78(9), 1268 - 74 Effect of feed withdrawal or challenge with Pasteurella multocida on growth, blood metabolites, circulating growth hormone, and insulin-like growth factor-I concentrations in eight-week-old turkeys; Anthony NB et al.; The daily effects of feed withdrawal or a bacterial disease (Pasteurella multocida; PM) challenge was studied in a slow-growing line of turkeys . The following groups (n = 6 birds/group) were sampled for up to 13 d: untreated control (CON), 4-d feed withdrawal followed by refeeding (FAST), a group that succumbed within the first 2 to 3 d after PM challenge (E-DEAD), a group that succumbed 8 to 9 d after PM challenge (L-DEAD), a group that survived the PM challenge (SUR), and a group treated with both PM challenge and 4-d feed withdrawal followed by refeeding (FAST/CHAL) . Daily feed intake and BW gains were markedly reduced in the E-DEAD and L-DEAD groups immediately and 3 d after PM challenge, respectively . Feed intake and BW gain between CON and SUR groups of turkeys were not different throughout the trial . The turkeys in the FAST group followed the expected feed withdrawal and refeeding patterns for feed intake and BW loss or gain . The FAST/CHAL turkeys consumed the minimal amount of feed to maintain BW after refeeding . Plasma uric acid sharply increased 1 d prior to death in both E-DEAD and L-DEAD groups of turkeys . Plasma uric acid also increased each consecutive day during fasting in the FAST and FAST/CHAL groups of turkeys . Plasma growth hormone was measured in only the CON and FAST groups and increased from about 40 to 85 ng/mL in the FAST group during fasting but returned to control levels within 1 d of refeeding . Circulating plasma insulin-like growth factor-I (IGF-I) decreased from about 17 to 5 ng/mL in the PM-challenged (E-DEAD, L-DEAD, and FAST/CHAL groups) and FAST groups . The concentration of IGF-I returned to prefeed withdrawal levels within 3 d of refeeding the FAST group of turkeys . It was concluded that 1) turkey poults that were not susceptible to the PM challenge generally maintained physiological functions at control bird levels, 2) susceptible turkey poults generally exhibited depressed feed intake and BW gains, and 3) poults challenged with both feed withdrawal and PM treatment responded differently than poults challenged with either feed withdrawal or challenge with PM . The depletion of energy intake and mobilization of energy stores in susceptible poults might have contributed to the rate at which PM caused the poults to die. Poult Sci, 1999 Sep, 78(9), 1263 - 7 Influence of body weight restriction in a body-weight-selected line of turkeys on response to challenge with Pasteurella multocida; Nestor KE et al.; Previous research has shown that a line (F) of turkeys selected long-term for increased 16-wk BW was more susceptible to challenge with washed Pasteurella multocida (PM) than a randombred control line (RBC2), the base population of the F line . Published research indicated that the mortality of the F line following challenge with PM was similar to that of two commercial sire lines . The purpose of the present study was to determine the influence of reducing BW of the F line to that of the RBC2 line by nutrient restriction on resistance to PM . Four challenge trials were conducted over a 2-yr period . The BW of a group of F line birds was restricted to that of the RBC2 line by limiting access to feed from 1 to 6 wk of age . The F line restricted birds and full-fed RBC2 and F line birds were challenged with a field isolate of washed PM (1.2x10(7) organisms/bird of capsular serogroup A and somatic serotype 3, 4) at 6 wk of age . Birds were checked twice daily for 14 d . Resistance to PM was measured by days to death of those that died and percentage mortality . The BW of the restricted group of the F line did not differ from full-fed RBC2 birds for males or females . In males, the restricted F line birds had similar mortality (48.0%) to the full-fed RBC2 line birds (44.3%), and the mortalities in both groups were significantly lower than that observed for the full-fed F line birds (81.3%) following challenge with PM . The mortality following challenge in females did not differ significantly among groups, even though mortality of the full-fed F line birds (64.1%) and restricted F line birds (63.3%) was more than 9% higher than that (54.2%) observed for the full-fed RBC2 line birds . Days to death was not a sensitive indicator of resistance to PM, as no differences among the three groups of birds were observed for either sex. Res Vet Sci, 1999 Oct, 67(2), 163 - 70 Ultrastructural pathology of the upper respiratory tract of rabbits experimentally infected with Pasteurella multocida A:3; Al-Haddawi MH et al.; Twenty-four 8 to 9 week-old Pasteurella multocida -free rabbits were divided into three equal groups, the first group was pretreated with hydrocortisone and inoculated intranasally with pasteurella multocida serotype A:3 . The second group was inoculated intranasally with P . multocida without hydrocortisone treatment . The third group was inoculated with phosphate buffered saline only and used as a control group . Pasteurella multocida was isolated from the nasal cavity of all infected rabbits in group 1 and 2 and from the trachea of seven rabbits in group 1 and five rabbits in group 2 . This study was conducted to observe the ultrastructural changes of the upper respiratory tract of hydrocortisone treated and non-treated rabbits infected with P . multocida serotype A:3 . The ultrastructural changes detected in infected rabbits were ciliary destruction and deciliation of the ciliated epithelial cells, cellular swelling, goblet cell hyperplasia and endothelial cell damage . Pasteurella multocida was observed attached to the degenerated cilia, microvilli and mucus . Pasteurella multocida infection was associated with inflammatory responses, which may have caused tissue damage . It is possible that hydrocortisone modulates the severity of infection as an immune suppressor and an inhibitor of goblet cell secretion . Microb Pathog, 1999 Oct, 27(4), 197 - 206 Protective capacity of the Pasteurella haemolytica transferrin-binding proteins TbpA and TbpB in cattle; Potter AA et al.; The transferrin-binding proteins TbpA and TbpB from Pasteurella haemolytica biotype A serotype 1 were tested for their ability to confer protection against experimental P . haemolytica infection when administered to calves in vaccine formulations containing one or both antigens . Vaccine groups included TbpB (single immunization), TbpB (two immunizations), TbpA, TbpA+TbpB and a placebo . All animals that received TbpB had measurable antibody titres against the antigen at the time of challenge, while those that received TbpA did not show an antibody response . The TbpA+TbpB group showed the best protection against experimental challenge . Protection correlated with anti-TbpB antibody levels . The enhanced protection in the TbpA+TbpB group suggests TbpA contributed to protection through the induction of a non-antibody-mediated immune response . Sera from the TbpB-immunized animals was cross-reactive with TbpBs from other P . haemolytica serotypes . Pediatr Nephrol, 1999 Oct, 13(8), 646 - 8 Capnocytophaga canimorsus peritonitis in a pediatric peritoneal dialysis patient; Chadha V et al.; Capnocytophaga canimorsus, a bacterium rarely encountered by clinicians, was responsible for the development of peritonitis in an 18-year-old white male on automated peritoneal dialysis following the puncture of his dialysis tubing by a domestic cat . Although more than 100 cases of septicemia caused by C . canimorsus have been reported, this is the first report of the organism causing peritonitis in a patient receiving peritoneal dialysis . Of interest, the patient had a prior episode of peritonitis secondary to Pasteurella multocida, also following transmission from the same cat. Avian Dis, 1999 Jul-Sep, 43(3), 549 - 52 Survey of pathogens and blood parasites in free-living passerines; Morishita TY et al.; To determine the disease prevalence of free-living passerines, 1709 passerines were sampled from 38 different field sites in Ohio . Choanal and cloacal swabs were collected from each bird and cultured for the presence of Pasteurella multocida, Salmonella spp., and Escherichia coli by standard microbiologic techniques . In addition, the serum from each bird was analyzed for the presence of antibodies to Mycoplasma gallisepticum, Mycoplasma synoviae, Newcastle disease virus, and avian influenza virus . A blood smear was also made to examine for the presence of blood parasites . Results indicated that the isolation of E . coli varied with bird species, with the European starling having a higher (21.4%) isolation of E . coli . Salmonella spp . were also isolated from these free-living passerines . Pasteurella multocida was not isolated from any of the sampled passerines . These birds did not have antibodies to M . gallisepticum, M . synoviae, Newcastle disease virus, or avian influenza virus . Blood parasites were not detected in any of the birds sampled. Vet Pathol, 1999 Sep, 36(5), 437 - 44 In situ expression of intercellular adhesion molecule-1 (ICAM-1) mRNA in calves with acute Pasteurella haemolytica pneumonia; Radi ZA et al.; The in situ expression of intercellular adhesion molecule-1 (ICAM-1) mRNA in normal and pneumonic lung tissues of Holstein calves with bovine leukocyte adhesion deficiency (BLAD) was compared with that of age-matched non-BLAD Holstein calves by in situ hybridization . Twenty-four Holstein calves (both BLAD and non-BLAD) were randomly assigned to one of two experimental groups and inoculated intrabronchially with Pasteurella haemolytica or pyrogen-free saline . Lung tissues were collected and fixed in 10% neutral formalin at 2 or 4 hours postinoculation (PI) . The expression and distribution of ICAM-1 mRNA in the different cell types of the lung tissue was detected by in situ hybridization with a 307-base-pair bovine ICAM-1 riboprobe . In lungs of both non-BLAD and BLAD saline-inoculated calves, ICAM-1 expression was present in epithelial cells but occurred in <30% of cells in bronchi, bronchioles, and alveoli . ICAM-1 expression in vascular endothelial cells was present in <30% of cells in pulmonary arteries and veins . The expression of ICAM-1 was significantly greater (>60% of cells) in bronchiolar and alveolar epithelial cells and pulmonary endothelial cells of arteries and veins in both BLAD and non-BLAD calves inoculated with P . haemolytica . Bronchiolar epithelium had the highest intensity of mRNA expression and highest percentage of cells that were stained, whereas bronchial epithelium had the lowest intensity and percentage of cells stained . Most alveolar macrophages and neutrophils in infected lungs also expressed ICAM-1 . ICAM-1 expression was generally increased in infected BLAD calves at 2 hours PI as compared with non-BLAD calves but not at 4 hours PI . The increased expression of ICAM-1 during acute P . haemolytica pneumonia in calves suggests that ICAM-1 is upregulated and may play a role in leukocyte infiltration . The extent of ICAM-1 expression in P . haemolytica-inoculated calves with BLAD was initially enhanced but otherwise similar to that in non-BLAD calves. Vet Pathol, 1999 Sep, 36(5), 397 - 405 Expression of inducible nitric oxide synthase in spontaneous bovine bronchopneumonia; Fligger JM et al.; The expression of inducible nitric oxide synthase (iNOS), major histocompatibility class II molecules (MHC-II), CD68, and the calcium-binding proteins S100A8 and S100A9 (also called MRP8 and MRP14, respectively) was assessed in lung tissues from cattle that succumbed to pneumonia . Expression patterns of these markers were related to the types of lung lesion . iNOS expression was only observed in lungs infected with Arcanobacterium pyogenes or Pasteurella haemolytica but not in lungs from cattle with subacute chronic interstitial pneumonia and acute interstitial pneumonia due to Escherichia coli infection . High levels of iNOS were expressed by cells (probably leukocytes) surrounding necrotic foci . Occasionally, iNOS was expressed by intraalveolar macrophages in viable parenchyma, by leukocytes within the airways, and by some chondrocytes in the supporting cartilage of bronchi . Cells expressing MHC-II were distributed relatively evenly throughout areas of inflammation and did not display any clear association with necrotic foci . Cell types expressing MHC-II included type II alveolar epithelial cells, spindle-shaped cells of the interstitium, cells in bronchus-associated lymphoid tissue, and leukocytes in lymph and blood vessels but largely excluded iNOS-positive cells . Likewise, CD68-positive cells were rarely positive for iNOS and were not confined to the areas surrounding necrotic tissue . As with MHC-II and CD68, there was little if any coexpression of iNOS and either of the S100 proteins tested . Thus, in cattle with necrotizing bronchopneumonia, iNOS-expressing cells were largely restricted to the cellular zone surrounding necrotic areas. J Zoo Wildl Med, 1999 Jun, 30(2), 285 - 92 Hemorrhagic septicemia in fallow deer (Dama dama) caused by Pasteurella multocida multocida; Eriksen L et al.; Four outbreaks of hemorrhagic septicemia caused by Pasteurella multocida multocida occurred in a population of 1,800 fallow deer (Dama dama) during 1992-1996 . A total of 340 fallow deer were submitted for postmortem examination . Pasteurellosis was diagnosed in 273 of 312 deer suspected of having septicemia . Pasteurella multocida was isolated from 257 animals, and the diagnosis was based on typical pathologic changes alone in the other 16 animals . Pasteurella multocida was isolated in pure culture from 219 of 248 samples of cerebrospinal fluid . Eighteen animals were observed moribund with severe depression, foamy nasal discharge, and respiratory distress, and 257 were found dead . Major clinical signs and pathologic changes included extensive swelling of the head and the neck and peracute or acute septic pneumonia, petechial and ecchymotic hemorrhages on serous membranes, and severely hemorrhagic adrenal glands and abomasum . Rhinitis and necrotic pharyngeal mucosae were common . Histologically, the most advanced lesions were in the nasal mucosa and pharynx . The swelling of the head and the neck arose from a diffuse cellulitis in the subcutaneous and intermuscular tissues . The earliest lesions in the lungs included large numbers of bacteria in the pulmonary capillaries, but various degrees of fibrinous exudation to the alveoli and infiltration with heterophils usually were observed. FEMS Microbiol Lett, 1999 Oct 1, 179(1), 161 - 7 The leukotoxin of Pasteurella haemolytica binds to beta(2) integrins on bovine leukocytes; Ambagala TC et al.; The putative receptor proteins of Pasteurella haemolytica leukotoxin were isolated from bovine polymorphonuclear neutrophil lysate by affinity chromatography on a leukotoxin-specific monoclonal antibody column to which the leukotoxin was pre-bound . SDS-PAGE of the purified proteins showed four bands at 180 kDa, 170 kDa, 150 kDa and 95 kDa, in addition to the expected 102-kDa leukotoxin band and a series of bands with molecular masses lower than 102 kDa representing the disintegrated leukotoxin . N-terminal amino acid sequencing of the 170-kDa band showed homology with human and murine CD11b . The purified proteins reacted specifically with monoclonal antibodies specific for CD11a, CD11b, CD11c (the alpha chains of beta(2) integrins), and CD18 (the beta chain of beta(2) integrins) . Pre-incubation of polymorphonuclear neutrophils with a monoclonal antibody specific for CD18 reduced the cytotoxicity of the leukotoxin to the cells . These results indicate that the leukotoxin binds to the beta(2) integrins on bovine leukocytes, very likely via CD18. J Wildl Dis, 1999 Jul, 35(3), 440 - 9 Antibodies against Pasteurella multocida in snow geese in the western Arctic; Samuel MD et al.; To determine if lesser snow geese (Chen caerulescens caerulescens) are a potential reservoir for the Pasteurella multocida bacterium that causes avian cholera, serum samples and/or pharyngeal swabs were collected from > 3,400 adult geese breeding on Wrangel Island (Russia) and Banks Island (Canada) during 1993-1996 . Pharyngeal swab sampling rarely (> 0.1%) detected birds that were exposed to P . multocida in these populations . Geese with serum antibody levels indicating recent infection with P . multocida were found at both breeding colonies . Prevalence of seropositive birds was 3.5% at Wrangel Island, an area that has no recorded history of avian cholera epizootics . Prevalence of seropositive birds was 2.8% at Banks Island in 1994, but increased to 8.2% during 1995 and 1996 when an estimated 40,000-60,000 snow geese were infected . Approximately 50% of the infected birds died during the epizootic and a portion of the surviving birds may have become carriers of the disease . This pattern of prevalence indicated that enzootic levels of infection with P . multocida occurred at both breeding colonies . When no avian cholera epizootics occurred (Wrangel Island, Banks Island in 1994), female snow geese (4.7%) had higher antibody prevalence than males (2.0%). J Biol Chem, 1999 Sep 10, 274(37), 26557 - 62 Molecular directionality of polysaccharide polymerization by the Pasteurella multocida hyaluronan synthase; DeAngelis PL; Hyaluronan (HA), a long linear polymer composed of alternating glucuronic acid and N-acetylglucosamine residues, is an essential polysaccharide in vertebrates and a putative virulence factor in certain microbes . All known HA synthases utilize UDP-sugar precursors . Previous reports describing the HA synthase enzymes from Streptococcus bacteria and mammals, however, did not agree on the molecular directionality of polymer elongation . We show here that a HA synthase, PmHAS, from Gram-negative P . multocida bacteria polymerizes the HA chain by the addition of sugar units to the nonreducing terminus . Recombinant PmHAS will elongate exogenous HA oligosaccharide acceptors to form long polymers in vitro; thus far no other HA synthase has displayed this capability . The directionality of synthesis was established definitively by testing the ability of PmHAS to elongate defined oligosaccharide derivatives . Analysis of the initial stages of synthesis demonstrated that PmHAS added single monosaccharide units sequentially . Apparently the fidelity of the individual sugar transfer reactions is sufficient to generate the authentic repeating structure of HA . Therefore, simultaneous addition of disaccharide block units is not required as hypothesized in some recent models of polysaccharide biosynthesis . PmHAS appears distinct from other known HA synthases based on differences in sequence, topology in the membrane, and putative reaction mechanism. Berl Munch Tierarztl Wochenschr, 1999 Jun-Jul, 112(6-7), 254 - 9 {Breath condensate--a medium obtained by a noninvasive method for the detection of inflammation mediators of the lung}; Reinhold P et al.; Collection of exhaled condensate (freezing of expired air under conditions of spontaneous breathing) is a non-invasive method permitting the collection of material originating from the lung and the lower respiratory tract so that it can be used for diagnostic examinations . In order to be able to evaluate the diagnostic evidence of exhaled condensate samples in cases of respiratory disease of the calf, leukotriene B4 (LTB4) in bovine exhaled condensate was determined . The influence of the breathing pattern and body temperature on the quantity of condensate to be collected was tested in a total of 49 exhaled condensate samples . It became obvious that the exhaled condensate quantity obtained per time unit is dependent on the ventilation volume per time unit (minute volume) . In exhaled condensate samples from 35 clinically healthy calves, LTB4 concentrations of less than 250 pg/mL exhaled condensate were detected . A total of 14 exhaled condensate samples from 7 calves was analyzed before and after experimental respiratory infection with Pasteurella multocida D . In parallel to the analysis of LTB4 in exhaled condensate, the lung function of the calves was examined by means of impulse oscilloresistometry . The increase of LTB4 in the exhaled condensate post infection correlated significantly (p < or = 0.05) with decreases of respiratory reactance . The determination of LTB4 concentrations in exhaled condensate seems to be suitable, in principle, for the detection of inflammations in the respiratory system of the calf . Further studies are needed for the evaluation of the diagnostic validity of the method. Antimicrob Agents Chemother, 1999 Sep, 43(9), 2138 - 43 Effects of enrofloxacin on porcine phagocytic function; Schoevers EJ et al.; The interaction between enrofloxacin and porcine phagocytes was studied with clinically relevant concentrations of enrofloxacin . Enrofloxacin accumulated in phagocytes, with cellular concentration/extracellular concentration ratios of 9 for polymorphonuclear leukocytes (PMNs) and 5 for alveolar macrophages (AMs) . Cells with accumulated enrofloxacin brought into enrofloxacin-free medium released approximately 80% (AMs) to 90% (PMNs) of their enrofloxacin within the first 10 min, after which no further release was seen . Enrofloxacin affected neither the viability of PMNs and AMs nor the chemotaxis of PMNs at concentrations ranging from 0 to 10 microg/ml . Enrofloxacin (0.5 microg/ml) did not alter the capability of PMNs and AMs to phagocytize fluorescent microparticles or Actinobacillus pleuropneumoniae, Pasteurella multocida, and Staphylococcus aureus . Significant differences in intracellular killing were seen with enrofloxacin at 5x the MIC compared with that for controls not treated with enrofloxacin . PMNs killed all S . aureus isolates in 3 h with or without enrofloxacin . Intracellular S . aureus isolates in AMs were less susceptible than extracellular S . aureus isolates to the bactericidal effect of enrofloxacin . P . multocida was not phagocytosed by PMNs . AMs did not kill P . multocida, and similar intra- and extracellular reductions of P . multocida isolates by enrofloxacin were found . Intraphagocytic killing of A . pleuropneumoniae was significantly enhanced by enrofloxacin at 5x the MIC in both PMNs and AMs . AMs are very susceptible to the A . pleuropneumoniae cytotoxin . This suggests that in serologically naive pigs the enhancing effect of enrofloxacin on the bactericidal action of PMNs may have clinical relevance. Eur J Biochem, 1999 Aug, 263(3), 695 - 701 31P and 13C nuclear magnetic resonance studies of metabolic pathways in Pasteurella multocida characterization of a new mannitol-producing metabolic pathway; Rager MN et al.; Glucose metabolism of Pasteurella multocida was examined in resting cells in vivo using 13C NMR spectroscopy, in cell-free extracts in vitro using 31P NMR spectroscopy and using enzyme assays . The NMR data indicate that glucose is converted by the Embden-Meyerhof and pentose phosphate pathways . The P . multocida fructose 6-phosphate phosphotransferase activity (the key enzyme of the Embden-Meyerhof pathway) was similar to that of Escherichia coli . Nevertheless, and in contrast to that of E . coli, its activity was inhibited by alpha glycerophosphate . This inhibition is consistent with the very low fructose 6-phosphate phosphotransferase activity found in cell-free extracts of P . multocida using a spectrophotometric method . The dominant end products of glucose metabolism were mannitol, acetate and succinate . Under anaerobic conditions, P . multocida was able to constitutively produce mannitol from glucose, mannose, fructose, sucrose, glucose 6-phosphate and fructose 6-phosphate . We propose a new metabolic pathway in P . multocida where fructose 6-phosphate is reduced to mannitol 1-phosphate by fructose 6-phosphate reductase . Mannitol 1-phosphate produced is then converted to mannitol by mannitol 1-phosphatase. Int J Antimicrob Agents, 1999 Aug, 12(3), 229 - 37 Postantibiotic and physiological effects of tilmicosin, tylosin, and apramycin at subminimal and suprainhibitory concentrations on some swine and bovine respiratory tract pathogens; Diarra MS et al.; The antimicrobial activity of tilmicosin, tylosin, and apramycin on some important gram-negative swine and bovine pathogens namely, Pasteurella multocida, Pasteurella haemolytica, Bordetella bronchiseptica, and Actinobacillus pleuropneumoniae were studied in vitro . The effect of minimal inhibitory concentrations (MICs) and sub-MICs (1/4, 1/2 MIC) on bacterial growth was evaluated . The presence of tilmicosin, tylosin and apramycin in the medium decreased the rate of growth of the bacterial strains tested using drug concentrations as low as 1/4 MIC . The postantibiotic effect (PAE) which is the suppression of optimal bacterial growth that persists after a short exposure (2 h) of microorganisms to an antibiotic was studied by exposure of bacteria to drugs at 1/4, 1/2, 1, 4 and 8 times MIC . The duration of PAEs increased with rising concentration for all drugs tested but at concentrations below the MIC, tilmicosin showed more significant PAEs than tylosin or apramycin against P . multocida and A . pleuropneumoniae . Tilmicosin and tylosin caused PAEs of up to 8 h when used at 8 times the MIC, while apramycin caused PAEs of up to 5 h when used at this concentration . Sub-MICs of either tilmicosin, tylosin, or apramycin had no effect on P . multocida dermonecrotic toxin production . However sub-MICs of tylosin, or apramycin significantly reduced the haemolytic activity of A . pleuropneumoniae and affected the capsular material production of this isolate and of one isolate of P . multocida (type A) . The in vitro effect of tilmicosin, tylosin, and apramycin (even when used at sub-MIC levels) on growth, production of capsular material, and haemolytic activity might impair the virulence of some of the microorganisms studied . In addition to the effects of these drugs on some putative virulence factors, we suggest that the strong PAEs caused by tilmicosin, tylosin, and apramycin may also contribute to the in vivo efficacy of these drugs. Infect Immun, 1999 Sep, 67(9), 4968 - 73 Molecular cloning of the Pasteurella haemolytica pomA gene and identification of bovine antibodies against PomA surface domains; Zeng H et al.; The gene (pomA) encoding PomA, an OmpA-like major outer membrane protein of the bovine respiratory pathogen Pasteurella haemolytica, was cloned, and its nucleotide sequence was determined . The deduced amino acid sequence of PomA has significant identity with the sequences of other OmpA family proteins . Absorption of three different bovine immune sera with whole P . haemolytica cells resulted in a reduction of bovine immunoglobulin G reactivity with recombinant PomA in Western immunoblots, suggesting the presence of antibodies against PomA surface domains. Microb Pathog, 1999 Sep, 27(3), 179 - 85 Use of TUNEL staining to detect apoptotic cells in the lungs of cattle experimentally infected with Pasteurella haemolytica; Leite F et al.; Lung sections taken from calves with experimental Pasteurella haemolytica respiratory infection exhibited increased numbers of TUNEL positive cells with time after challenge . This finding suggests that P . haemolytica, or toxins and other components released by the organism, induces apoptosis in bovine leukocytes in vivo . By so doing this might impair host defense and contribute in part to the severe pneumonia that characterizes bovine pasteurellosis . Antimicrob Agents Chemother, 1999 Aug, 43(8), 1988 - 92 Pharmacokinetics of enrofloxacin and danofloxacin in plasma, inflammatory exudate, and bronchial secretions of calves following subcutaneous administration; McKellar Q et al.; Enrofloxacin (2.5 mg/kg of body weight) and danofloxacin (1.25 mg/kg) were administered subcutaneously to ruminating calves (n = 8) fitted with subcutaneous tissue cages . Concentrations of enrofloxacin, its metabolite ciprofloxacin, and danofloxacin in blood (plasma), tissue cage exudate (following intracaveal injection of 0.3 ml of 1% {vol/wt} carrageenan), and bronchial secretions were measured by high-performance liquid chromatography (HPLC) and microbiological assay (enrofloxacin plus ciprofloxacin and danofloxacin) . Mean maximum concentrations (C(max)) +/- standard deviations of enrofloxacin (0.24 +/- 0.08 microg/ml), ciprofloxacin (0.11 +/- 0.03 {total, 0.34 +/- 0.10} microg/ml), and danofloxacin (0.23 +/- 0.05 microg/ml) were detected in the plasma of calves by HPLC . The C(max) were 0.49 +/- 0.17 microg/ml (enrofloxacin equivalents) and 0.24 +/- 0.03 microg/ml (danofloxacin) when they were measured by microbiological assay . Mean C(max) in exudate (HPLC) were 0.18 +/- 0.07 microg/ml (enrofloxacin), 0.10 +/- 0.04 microg/ml (ciprofloxacin), 0.27 +/- 0.09 microg/ml (enrofloxacin plus ciprofloxacin), and 0.19 +/- 0.05 microg/ml (danofloxacin), and concentrations in exudate exceeded those in plasma from 8 h (enrofloxacin and ciprofloxacin) or 6 h (danofloxacin) after drug administration . The C(max) were 0.34 +/- 0.09 microg/ml (enrofloxacin equivalents) and 0.22 +/- 0.04 microg/ml (danofloxacin) in exudate when they were measured by the microbiological assay . The maximum mean concentration achieved in bronchial secretions (HPLC) were 0.07 +/- 0.04 microg/ml (enrofloxacin), 0.04 +/- 0.07 microg/ml (ciprofloxacin), 0.10 +/- 0 . 05 microg/ml (enrofloxacin plus ciprofloxacin), and 0.12 +/- 0.09 microg/ml (danofloxacin) . The maximum mean concentration in bronchial secretions from a limited number of animals from which samples were available for microbiological assay were 0.27 +/- 0.11 microg/ml (n = 4 {enrofloxacin equivalents}) and 0.14 +/- 0.02 microg/ml (n = 3 {danofloxacin}) . With predictive models of efficacy (C(max)/MIC and area under the concentration-time curve/MIC ratios in plasma) for Pasteurella multocida (MIC of enrofloxacin, 0.06 microg/ml {24}; MIC of danofloxacin, 0.06 microg/ml {6}), enrofloxacin produced scores of 8.17 and 52.00, respectively, compared to those of danofloxacin, which were 4.02 and 23.05, respectively . With the dosing rates recommended in some markets by manufacturers, enrofloxacin and danofloxacin achieved concentrations above the MICs for important pathogenic organisms in plasma, tissue cage exudate, and bronchial secretion . Since fluoroquinolones display concentration-dependent activities, C(max)/MIC ratios may be critical to efficacy . In the United States enrofloxacin is currently the only fluoroquinolone licensed for food animals and dosages for acute respiratory disease are 2.5 to 5 mg/kg for 3 days or 7.5 to 12 . 5 mg/kg once . The higher dosages on a single occasion are likely to confer C(max)/MIC ratios that are associated with greater clinical efficacy. Vet Q, 1999 Jun, 21(3), 99 - 104 Sensitivity testing of veterinary pathogens with a semi-automatic image analysis system compared with tablet diffusion and agar dilution tests; Mevius DJ et al.; Recently a commercial computer-controlled image analysis system (IAS) was introduced to measure automatically the diameters of inhibition zones in the agar diffusion test . However, there is little information on the precision of this method . In the present study clinical isolates of Salmonella spp . (N = 104), Escherichia coli (N = 100), Pasteurella spp . (N = 99), Actinobacillus pleuropneumoniae (N = 85), porcine streptococci (N = 100), and Staphylococcus aureus (N = 95) were tested in the agar diffusion test, using nineteen different antibiotics in tablets . All inhibition zone diameters were first measured by a laboratory technician and then by the IAS . Although the zone diameters of all bacteria-antibiotic combinations measured by the IAS and those measured by the laboratory technician showed a significant positive correlation, the size of the inhibition zone diameters measured by the technician and the IAS differed significantly in 59% of the combinations . However, these differences were very small and may have no clinical relevance . The IAS was also used to calculate minimum inhibitory concentrations (MIC values) from the zone diameters . In 82% of the bacteria-antibiotic combinations MIC values calculated by the IAS showed a significant positive correlation with MIC values obtained with the reference agar dilution test . However, in 92% of the bacteria-antibiotic combinations, the calculated MIC values differed significantly from the reference values . In some cases these differences were so large that they could be of clinical relevance . The IAS was unable to measure the diameter of inhibition zones of porcine streptococci properly, due to poor contrast . We concluded that when tablets are used as antibiotic carriers the IAS accurately measures the diameter of inhibition zones for bacteria species that give good contrast between the agar and bacterial growth . MIC values determined with the IAS were only indicative of those determined with the reference agar dilution test. Kansenshogaku Zasshi, 1999 Jun, 73(6), 623 - 5 {Two cases of pasteurellosis accompanied by exudate with semen-like odor from the wound}; Arashima Y et al.; We encountered two cases of Pasteurella multocida subsp . septica isolation from exudates with seminal fluid-like odor from dog scratch and cat bite . Case 1: A 78-year-old male who had been diagnosed as having diabetes mellitus five years ago was scratched by the claw of a pet dog (Pekinese) on the back of the right hand . Since inflammation ascended to the arm, the patient visited Nihon University Itabashi hospital for a medical examination . Case 2: A 51-year-old female without a specific past history other than hyperlipidemia was bitten by a pet cat at the medical and lateral sides of the left carpus . The patient immediately opened the wound and washed it with tap water, followed by disinfection using a non-iodine disinfectant at home . Two hours later, the patient felt an unpleasant sensation and smelled a seminal fluid-like odor at the wound . The next morning, the entire left arm swelled and pain worsened, then the patient sought medical attention . The patients were treated with antibiotics and the wound completely healed on the 16 days from on set in Case 1 and on the 10 days from onset in Case 2 . From these two cases, Pasteurella multocida subsp . septica was isolated from the exudate, suggesting that when wounds caused by animals smell like seminal fluid, the wound is infected with Pasteurellae . This finding may be an important clue for differentiation in clinical diagnosis. Infect Immun, 1999 Aug, 67(8), 3768 - 72 Role of phospholipase D in Pasteurella haemolytica leukotoxin-induced increase in phospholipase A(2) activity in bovine neutrophils; Wang Z et al.; The effects of Pasteurella haemolytica leukotoxin (LKT) on the activity of phospholipase D (PLD) and the regulatory interaction between PLD and phospholipase A(2) (PLA(2)) were investigated in assays using isolated bovine neutrophils labeled with tritiated phospholipid substrates of the two enzymes . Exposure of {(3)H}lysophosphatidylcholine-labeled neutrophils to LKT caused concentration- and time-dependent production of phosphatidic acid (PA), the product of PLD . LKT-induced generation of PA was dependent on extracellular calcium . Both production of PA and metabolism of {(3)H}-arachidonate ({(3)H}AA)-labeled phospholipids by PLA(2) were inhibited when ethanol was used to promote the alternative PLD-mediated transphosphatidylation reaction, resulting in the production of phosphatidylethanol rather than PA . The role of PA in regulation of PLA(2) activity was then confirmed by means of an add-back experiment, whereby addition of PA in the presence of ethanol restored PLA(2)-mediated release of radioactivity from neutrophil membranes . Considering the involvement of chemotactic phospholipase products in the pathogenesis of pneumonic pasteurellosis, development and use of anti-inflammatory agents that inhibit LKT-induced activation of PLD and PLA(2) may improve therapeutic management of the disease. Vet Microbiol, 1999 Jun 15, 67(2), 91 - 7 Bovine CD18 identified as a species specific receptor for Pasteurella haemolytica leukotoxin; Li J et al.; Ligand blotting of Pasteurella haemolytica leukotoxin (LKT) susceptible BL3 bovine lymphoma cell membranes with LKT detected two putative receptors with Mr of 95 and 100 kDa, whereas no LKT binding to membrane proteins was detected for LKT non-susceptible human leukemic cells . Anti-bovine CD18 and CD11a/CD18 mAb recognized 95 and 100kDa bands from BL3 cell membranes . CD18 isolated from BL3 cell membranes bound LKT . Pre-incubation of BL3 cells with anti-bovine CD18 or CD11a/CD18 mAb caused partial inhibition of LKT-induced leukolysis . Therefore, we propose that bovine CD18 acts as a species-specific leukocyte receptor for P . haemolytica LKT. Am J Vet Res, 1999 Jul, 60(7), 853 - 9 Evaluation of commercially available Escherichia coli J5 bacterin as protection against experimental challenge with Pasteurella multocida in rabbits; Ruble RP et al.; OBJECTIVE: To evaluate the ability of commercially available Escherichia coli J5 bacterin to protect rabbits from experimental challenge with Pasteurella multocida . ANIMALS: 40 P multocida-free New Zealand White rabbits . PROCEDURES: Rabbits were assigned to 1 of 4 groups of 10 rabbits each . Three of the groups were inoculated SC with J5 bacterin at 8 weeks old . Inoculation was repeated 3 and 6 weeks later . The fourth group was not inoculated and served as controls . Groups 1, 2, and 3 were given 10(9), 10(8), and 10(7) colony forming units (CFU), respectively . Response was monitored by titer assessment, using an E coli J5 antigen capture ELISA . Five weeks after the last inoculation, all rabbits were challenged with P multocida and observed for an additional 5 weeks . Clinical, hematologic, serologic, culture, and necropsy data were collected . RESULTS: Inoculation of rabbits with 10(9) CFU of E coli J5 bacterin-induced titers that were significantly greater than titers of rabbits vaccinated with 10(8) or 10(7) CFU or those in controls . The incidence of acute bacteremia was lower in rabbits with high titers . At necropsy, prevalence of lesions typical of P multocida was not significantly different among groups . Prevalence of histologic lesions was also not significantly different among groups . CONCLUSIONS AND CLINICAL RELEVANCE: Although the bacterin induced considerable antibody response and possibly reduced the rate of bacteremia, antibodies were not protective against long-term colonization or infection of the frontal sinuses or tympanic bullae by the challenge strain of P multocida . This bacterin in its currently available form is unlikely to aid in reducing the prevalence of pasteurellosis in rabbits. Avian Dis, 1999 Apr-Jun, 43(2), 279 - 85 Virulence of raptor-origin Pasteurella multocida in domestic chickens; Aye PP et al.; Pasteurella multocida belonging to somatic serotype 1 and capsular type A has been known to cause avian cholera in domestic poultry . Pasteurella multocida serotype 1 has also been isolated from raptorial birds . However, the capsular type for these raptorial isolates remains unknown . Moreover, the virulence of these raptorial isolates for domestic poultry has not been determined . The objectives of this study were to determine the capsular type of raptorial P . multocida serotype 1 isolates and to determine if these isolates were virulent for domestic chickens . Study chickens were inoculated with one of three P . multocida isolates . Isolate WESO-1 was obtained from a western screech owl (Otus kennicottii) and isolates RTHA-2 and RTHA-4 were isolated from two red-tailed hawks (Buteo jamaicensis) . These isolates were given by either the oral, intravenous, or intraocular route . Control birds were given brain-heart infusion broth . The capsular serotypes of three isolates were also determined . The RTHA-2 and RTHA-4 isolates belonged to P . multocida capsular type A . The WESO-1 isolate belonged to capsular type F . Results also demonstrated that, for the isolates examined, the intraocular route did not cause mortality in chickens . There was mortality in all groups for the intravenous route . However, various mortality patterns were observed when P . multocida was given orally for the three different isolates . The RTHA-4 isolate (serotype 1:A) was the most virulent for domestic chickens . The WESO-1 isolate (serotype 1:F) was the least virulent for chickens among the raptorial isolates examined. Clin Diagn Lab Immunol, 1999 Jul, 6(4), 617 - 20 Conservation of expression and N-terminal sequences of the Pasteurella haemolytica 31-kilodalton and Pasteurella trehalosi 29-kilodalton periplasmic iron-regulated proteins; Tabatabai LB et al.; This study examined the conservation of expression of a 31-kDa iron-regulated protein by serotypes of Pasteurella haemolytica and Pasteurella trehalosi associated with pasteurellosis of cattle and sheep . A polyclonal antibody prepared against the purified 31-kDa periplasmic iron-regulated protein from P . haemolytica serotype A1 showed that all P . haemolytica serotypes expressed similar 31-kDa proteins with identical N-terminal sequences, whereas P . trehalosi serotypes expressed immunologically different 29-kDa proteins with a different N-terminal sequence . Antibody to the 31-kDa iron-regulated protein was a useful tool to distinguish similarities and differences of the iron-regulated proteins of P . haemolytica and P . trehalosi. Appl Environ Microbiol, 1999 Jul, 65(7), 2942 - 6 16S rRNA gene sequence analysis of Photobacterium damselae and nested PCR method for rapid detection of the causative agent of fish pasteurellosis; Osorio CR et al.; The causative agent of fish pasteurellosis, the organism formerly known as Pasteurella piscicida, has been reclassified as Photobacterium damselae subsp . piscicida on the basis of 16S rRNA gene sequence comparisons and chromosomal DNA-DNA hybridization data; thus, this organism belongs to the same species as Photobacterium damselae subsp . damselae (formerly Vibrio damselae) . Since reassignment of P . damselae subsp . piscicida was based on only two strains, one objective of the present work was to confirm the taxonomic position of this fish pathogen by sequencing the 16S rRNA genes of 26 strains having different geographic and host origins . In addition, a nested PCR protocol for detection of P . damselae based on 16S rRNA was developed . This PCR protocol was validated by testing 35 target and 24 nontarget pure cultures, and the detection limits obtained ranged from 1 pg to 10 fg of DNA (200 to 20 cells) . A similar level of sensitivity was observed when the PCR protocol was applied to fish tissues spiked with bacteria . The PCR approach described in this paper allows detection of the pathogen in mixed plate cultures obtained from asymptomatic fish suspected to be carriers of P . damselae subsp . piscicida, in which growth of this bacterium cannot be visualized . Our results indicate that the selective primers which we designed represent a powerful tool for sensitive and specific detection of fish pasteurellosis. J Vet Med Sci, 1999 May, 61(5), 565 - 7 Effects of the lipopolysaccharide-protein complex and crude capsular antigens of Pasteurella multocida serotype A on antibody responses and delayed type hypersensitivity responses in the chicken; Maslog FS et al.; The effects of the lipopolysaccharide-protein complex (LPS) and crude capsular antigen (CCA) prepared from Pasteurella multocida serotype A isolated from a duck in the Philippines, on antibody responses to sheep red blood cells (SRBC) and Brucella abortus (BA) and delayed type hypersensitivity (DTH) responses to bovine serum albumin (BSA) in the chickens were studied . Chickens injected subcutaneously with LPS and CCA at 1 and 2 weeks of age and immunized intravenously with the mixed antigens of SRBC and BA, at 3 and 4 weeks of age showed significantly increased antibody responses against both SRBC and BA, when evaluated at 7 days after each immunization . In addition, these chickens sensitized intramuscularly with the emulsion of BSA in complete Freund's adjuvant at 5 weeks of age, and then injected into the wattle with BSA at 7 weeks of age also showed significantly increased DTH responses against BSA, when evaluated at 24 and 48 hr after challenge . These results indicate that LPS and CCA of P . multocida serotype A have a property enhancing humoral and cell-mediated immune responses. Presse Med, 1999 May 22-29, 28(19), 1014 - 6 {Pasteurella multocida pneumonia in a lupus patient: microbial identification with alveolar lavage fluid on blood culture medium}; Hazouard E et al.; BACKGROUND: Pasteurella multocida pneumonia mainly occurs in immunodepressed patients . Microbiological proof is difficult to obtain . CASE REPORT: A 36-year-old woman with systemic lupus erythematosus treated with cyclophosphamide and corticosteroids developed pneumonia . She was given amoxicillin-clavulanate . Bronchioalveolar lavage fluid cultures on gelose were negative but Pasteurella multocida grew on blood culture medium . DISCUSSION: Although the direct examination of bronchioalveolar lavage fluid demonstrated Gram negative coccobacilli, gelose cultures were negative, probably because of prior antibiotic therapy . The causal pathogen was only identified when BAL fluid was seeded on blood culture medium, allowing susceptibility tests and subsequent early adaptation of antibiotic therapy . This technique can be helpful in identifying the casual pathogen in microbial pneumonia. Zentralbl Veterinarmed B, 1999 May, 46(4), 241 - 7 Serotypes of Pasteurella haemolytica and Pasteurella trehalosi isolated from farm animals in Hungary; Fodor L et al.; The biochemical and serological characteristics of 486 P . haemolytica and 31 P . trehalosi strains (517 in total) isolated from different lesions of cattle, sheep, goats, pigs and poultry were examined . A total of 476 P . haemolytica strains (97.9%) showed the characteristics typical of the former biotype A of P . haemolytica, while 10 isolates (2.1%), all from poultry, could not be biotyped . A total of 481 strains (93.0%) could be assigned to one of the 17 serotypes of P . haemolytica-P . trehalosi and 36 strains (7.0%) could not . The majority (83.6%) of the cattle isolates were serotypes A1 and A2 . Among strains isolated from sheep all serotypes of P . haemolytica could be identified with the exception of A14, but serotypes A1, A2, A6, A8 and A5 were the most frequent . The overwhelming majority (94%) of the caprine isolates were A2, other serotypes occurred only sporadically . The pig isolates, which could be isolated only very rarely, represented different serotypes, while none of the 10 strains isolated from poultry could be biotyped or serotyped. Dtsch Tierarztl Wochenschr, 1999 May, 106(5), 207 - 9 Serotypes and electrophoretic protein profiles of Pasteurella haemolytica isolated from pneumonic ovine lungs; Diker KS et al.; Serotypes and SDS-PAGE protein profiles of P . haemolytica isolated from pneumonic ovine lungs were investigated . Of 268 P . haemolytica isolates, 232 (86.6%) were serotypable . A total of 12 serotypes were recognized in 20 different geographic origins of central Turkey . The most common serotype was A2, followed by A7, A1 and T4 . Serotypes A13, A14, A16 and T15 could not be detected . In SDS-PAGE, marked differences between major bands of biotype A and T strains were found . In numerical analysis of protein profiles, biotype A and T strains were separated at 58% similarity level . Biotype A isolates produced a cluster at 80% similarity level, and biotype T isolates at 92% similarity level . No single cut off level was able to discriminate between each serotype studied and isolates could not be clustered on the basis of their geographic origins. J Bacteriol, 1999 Jun, 181(12), 3845 - 8 Cloning and characterization of the gene encoding Pasteurella haemolytica FnrP, a regulator of the Escherichia coli silent hemolysin sheA; Uhlich GA et al.; A Pasteurella haemolytica A1 gene was identified from a recombinant library clone that expressed hemolysis in host Escherichia coli cells . The gene, designated fnrP, had sequence identity to E . coli fnr, a global transcriptional regulator of genes required for conversion to anaerobic growth . FnrP complemented anaerobic deficiencies of a fnr-null mutant strain of E . coli and increased expression of the Fnr-dependent, anaerobic terminal reductase gene, frdA . FnrP was purified, identified by immunoblotting, and shown to be nonhemolytic . When FnrP was expressed in E . coli DeltasheA, a null mutant of the cryptic hemolysin SheA, the transformants were nonhemolytic, indicating that FnrP activates this silent hemolysin. Curr Microbiol, 1999 May, 38(5), 268 - 72 Derivation of extracellular polysaccharide-deficient variants from a serotype A strain of Pasteurella multocida; Champlin FR et al.; The production of serotype A extracellular polysaccharide is thought to be associated with expression of an approximately 40-kDa lipoprotein (P1p-40) present on the outer surface of Pasteurella multocida strains of avian origin . The tendency of certain strains to undergo colonial dissociation concomitantly with serial passaging on laboratory growth media was exploited to derive two variant strains exhibiting the capsule-deficient phenotype from a heavily capsulated parental strain . Assessments of colonial consistency, iridescence, gentian violet binding, and hyaluronidase sensitivity were consistent with cellular observations indicating little or no capsulation of derivative strains . Fluorographic analysis of electrophoretically resolved cellular lipoproteins labeled with {3H}-palmitate revealed capsular loss occurred with a concomitant diminution of P1p-40 production in the variant strains . In contrast, a phenotypically stable strain that did not undergo colonial dissociation under identical conditions exhibited no decrease in P1p-40 content . This work provides a model system for investigating the role of extracellular polysaccharide in the cell surface physiology and pathogenicity of P . multocida . The present results strongly support the notion that P1p-40 is associated with serotype A capsular material and suggest coordinate regulation of their biosynthesis. Antimicrob Agents Chemother, 1999 Jun, 43(6), 1469 - 74 Linezolid activity compared to those of selected macrolides and other agents against aerobic and anaerobic pathogens isolated from soft tissue bite infections in humans; Goldstein EJ et al.; Linezolid was tested against 420 aerobes and anaerobes, including 148 Pasteurella isolates, by an agar dilution method . Linezolid was active against all Pasteurella multocida subsp . multocida and P . multocida subsp . septica isolates and most Pasteurella canis, Pasteurella dagmatis, and Pasteurella stomatis isolates . The MIC was </=2 microg/ml for staphylococci, streptococci, EF-4b, Weeksella zoohelcum, Fusobacterium nucleatum, other fusobacteria, Porphyromonas spp., Prevotella spp., peptostreptococci, and almost all Bacteroides tectum isolates. Microb Pathog, 1999 Jun, 26(6), 325 - 31 Cytokine profiles following interaction between bovine alveolar macrophages and Pasteurella haemolytica; Morsey MA et al.; Pasteurella haemolytica is a gram negative bacterium frequently isolated from the lungs of calves suffering from a fibrinous pneumonic condition known as shipping fever . To understand the pathogenesis of this disease, we investigated the induction of cytokin gene expression in cultures of bovine alveolar macrophages (BAM) stimulated with heat-killed P . haemolytica . Northern blot analysis of total RNA showed that P . haemolytica induced early, abundant, and consistent synthesis of IL-1, TNF-alpha, and IL-8 mRNA . Cytokine mRNAs were detected within 1 hr post-stimulation with heat-killed P . haemolytica . IL-1 and IL-8 mRNA accumulated to high levels with increase in stimulation time, whereas TNF-alpha mRNA clearly declined by 4 and 8 h post stimulation . IL-1, TNF-alpha, and IL-8 proteins were also secreted into the culture medium of BAM stimulated with heat-killed P . haemolytica . All three proteins were detected at high levels 8 and 12 h post stimulation with P . haemolytica . BAM cells treated with bovine interferon-alpha and then stimulated with P . haemolytica produced higher amounts of IL-1, IL-8 and TNF-alpha proteins compared to BAM stimulated with P . haemolytica alone . These findings demonstrate the powerful and selective induction of cytokine mRNA and protein synthesis in BAM stimulated with heat-killed P . haemolytica and may explain certain aspects of shipping fever pathogenesis . Infect Immun, 1999 Jun, 67(6), 2920 - 7 Lipopolysaccharide complexes with Pasteurella haemolytica leukotoxin; Li J et al.; The presence of lipopolysaccharide (LPS) in gram-negative bacterial repeats-in-toxin (RTX) toxin preparations, as well as the harsh conditions required to remove it, suggests that LPS may complex with RTX toxins . Concentrated culture supernatant (CCS) preparations of the RTX toxin Pasteurella haemolytica leukotoxin (LKT) contained LKT and LPS as the most prominent components, with LKT and LPS constituting approximately 30 and 50% of the density of the silver-stained fraction on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), respectively . CCS LKT contained 3.69 +/- 0.46 mg of LPS per mg of protein, which was estimated to indicate an LPS/LKT molar ratio of approximately 60:1 . Subjection of the CCS LKT to preparative SDS-PAGE resulted in separation of LPS from LKT as detected by silver-stained analytical SDS-PAGE; however, the LKT fraction (SDS-PAGE LKT) contained significant endotoxin activity as detected by the Limulus amebocyte lysate assay . Subjection of the SDS-PAGE LKT to a second preparative SDS-PAGE run resulted in a reduction of the LPS/LKT molar ratio to 1:20 . The target cell specificity of LKT for bovine leukocytic cells was retained by the SDS-PAGE LKT, and isolated LPS at comparable concentrations to that in CCS LKT exhibited no leukolytic activity . Addition of isolated LPS back to SDS-PAGE LKT resulted in reconstitution of an LPS-LKT complex . Immediately following reconstitution of the LPS-LKT complex, there was minimal change in leukolytic activity of the complex, but following 9.5 h at temperatures from -135 to 37 degrees C, the LPS-LKT complex exhibited increased leukolytic activity and thermal stability compared to SDS-PAGE LKT . Therefore, it appears that LPS complexes with LKT, resulting in enhanced and stabilized leukolytic activity. J Emerg Med, 1999 May-Jun, 17(3), 445 - 8 Pasteurella multocida meningitis and septic arthritis secondary to a cat bite; Layton CT; Animal bites are seen almost daily in the emergency department, and the majority heal without complication . Pasteurella multocida is frequently the causative organism of localized wound infections and cellulitis in this patient population . P . multocida infection is usually associated with close contact with pets, such as dogs and cats, that harbor this organism as normal oral flora . Meningitis and septic arthritis are very rare sequelae of P . multocida infection . This case report presents a patient with P . multocida bacteremia, meningitis, and septic arthritis developing together as a complication of a cat bite. Zentralbl Veterinarmed B, 1999 Apr, 46(3), 145 - 50 Clinical and microbiological study of an otitis media outbreak in calves in a dairy herd; Yeruham I et al.; On a dairy farm, otitis media was diagnosed in 64 suckler calves (21.8%) during a study period of 2 years, and in 10 calves (3.4%) in the third year . The inflammation was unilateral in 63 and bilateral in 11 calves . The affected calves were dull, lacked appetite, were pyrexic and displayed drooping ear or ears and tilted heads with purulent discharge exuding from the external ear canal . Of the affected animals, 56 (87.5%) were aged between 3 and 8 weeks . Morbidity was higher during the calving season and during the autumn and winter months (October-December) . Pasteurella haemolytica was isolated from 21 (32.8%), P . multocida from 20 (31.2%), Actinomyces pyogenes from 11 (17.2%) and Streptococcus pneumoniae from three (4.7%) of the clinically affected calves only during the first two study years . The exudate of the acute ear infections contained, in addition to Pasteurella spp., various bacteria and yeasts . Most of these bacteria were isolated from healthy ears as well, and are likely to be part of the normal ear flora . On the other hand most of the yeasts were isolated from otitic calves . After a short course of an appropriate treatment infections healed in all cases . Possible preventive measures are discussed. J Med Microbiol, 1999 Mar, 48(3), 279 - 86 Genomic DNA restriction site heterogeneity in bovine Pasteurella multocida serogroup A isolates detected with an rRNA probe; Dabo SM et al.; A total of 81 Pasteurella multocida isolates from healthy and diseased dairy and beef cattle originating from various geographical locations was examined by rRNA gene restriction site polymorphism analysis (ribotyping), restriction endonuclease analysis (REA), SDS-PAGE analysis of whole-cell (WCP) and outer-membrane (OMP) proteins, and capsule and somatic serotyping . Bacterial strains were isolated from nose, lung and in one