Clin Microbiol Infect, 2002 Jan, 8(1), 55 - 7 Pasteurella gallinarum neonatal meningitis; Ahmed K et al.; A 4-day-old baby weighing 1.7 kg was admitted to the neonatal intensive care unit of Ga-Rankuwa Hospital, Pretoria, with a history of apneic attacks . On examination there was an umbilical sepsis and the neonate was septicemic . The baby had been delivered at home and the umbilical cord had been cut by the grandmother using unclean scissors and chimney soot applied to the umbilical stump . On admission, a septic screen was done and antibiotic treatment was started with penicillin and amikacin . The investigations showed that the baby was slightly anemic, with hemoglobin levels of 10.0 g/dL (14.9-23.7 g/dL), and a pure growth of a Gram-negative bacillus was obtained from the cerebrospinal fluid, blood culture and suprapubic aspirate urine specimens . The Gram-negative bacillus was catalase and oxidase positive and it was identified as Pasteurella gallinarum . Antimicrobial profiling showed the organism to be susceptible to penicillin, cefotaxime, gentamicin and amikacin . Despite having received antimicrobial agents to which the etiological agent was susceptible, the neonate died within 5 days of admission . The cause of death was postulated to be due to overwhelming sepsis which resulted in septic shock. Handchir Mikrochir Plast Chir, 2002 Jan, 34(1), 22 - 9 {Treatment of bites to the hand and wrist--is the primary antibiotic prophylaxis necessary?}; Rothe M et al.; Animal bites make up a large proportion of the injuries treated in an emergency department . Due to the type of injury, the variety of wounds and the contamination with aerobic and anaerobic organisms, they deserve special attention.In this study, we reviewed 98 patients (55 male/43 female) with bite wounds to the hand and wrist treated between 1995 and 2000 . They were either treated conservatively (n = 65) or surgically (n = 33) depending on the clinical findings . In 18 of 33 cases, the reason for surgical treatment was an infection . A primary antibiotic prophylaxis, usually with cephalosporines, was administered in 47 of 98 cases . Results were analysed retrospectively.An infection developed in 32 patients . In six of these patients, an infection developed despite primary antibiotic prophylaxis . Operative treatment became necessary in four of these six cases . Twenty-six of 32 patients were treated without primary antibiotic prophylaxis . Surgical treatment was required in around half (n = 14) of these patients, while the other 12 patients were treated conservatively with antibiotic therapy.Twenty-one of 26 patients presented with bite wounds that were already infected . Microbiological examination revealed a variety of microbes, usually a mixed infection with Pasteurella multocida was found . All organisms were susceptible to treatment with second or third generation cephalosporines.A total of 15 patients had to be operated due to deeper injuries to the bone, extensive soft-tissue injury, or because of injury to a tendon and the tendon sheath . In most patients, a good to acceptable functional result was achieved.Primary antibiotic prophylaxis does not prevent the development of infection . Nevertheless, because of the inherently high infection risk associated with bite wounds to the hand and wrist, prophylaxis should be carried out . In case of severe damage to the soft tissue or signs of infection, early surgical therapy should be considered. Reprod Domest Anim, 2001 Oct, 36(5), 247 - 56 Clinical and bacteriological aspects on the use of oxytetracycline and flunixin in primiparous cows with induced retained placenta and post-partal endometritis; Konigsson K et al.; Retention of the fetal membranes and post-partal endometritis (RFM) are common problems in dairy cows . Treatment often includes manual removal of the placenta in combination with antibiotic treatment . Earlier studies have shown that cows with endometritis post-partum have a strong tendency to recover spontaneously . The present study focused on treatments of post-partal endometritis with the prostaglandin synthesis inhibitor, flunixin (F) either alone or combined with oxytetracycline (T) . The study was conducted in two experiments, using 12 primiparous cows in each . As a model for RFM, premature parturition was induced in late pregnant heifers by injecting PGF2alpha (25 mg i.m.) twice with a 24 h interval . In each experiment the cows were set into four groups and treated with either T (10 mg/kg BW i.m . once daily), F (2.2 mg/kg BW p.o . twice daily), a combination of T and F (dosage, as above) or conservatively (group 0, no drugs) . The treatment periods lasted from days 11-14 post-partum in experiment I (groups T1, F1, TF1 and 0) and from days 3-6 post-partum in experiment 2 (groups T2, F2, TF2 and 0) . Jugular vein blood samples were collected for analyses of flunixin and total white blood cells . Uterine biopsies were collected twice weekly for investigation of endometrial microbiology . Rectal palpation and ultrasonographic examinations were performed three times weekly for investigations of uterine involution and ovarian activity . No attempts were made to remove the placentas manually . The experiment lasted until day 56 post-partum . The induction of parturition was successful in all heifers and 22 of 24 animals had RFM . All RFM cows had bacterial endometritis . The predominant bacteria were Escherichia coli alpha-haemolytic streptococci, Fusobacterium necrophorum, Arcanobacterium (Actinomyces) pyogenes, Bacteroides spp., Pasteurella spp . and Proteus spp . Fusobacterium necrophorum and A . pyogenes could be isolated for 3-5 weeks post-partum and E . coli Pasteurella and Proteus could be isolated for 2-3 weeks post-partum . Animals treated with tetracycline after placental shedding (T1 and TF1) had a more rapid recovery from infections with A . pyogenes and F . necrophorum than animals that were not treated with tetracycline . No other genera were affected . Antibiotic treatment before placental shedding (T2 and TF2) did not shorten the uterine infection but altered the bacterial flora, seen as an overgrowth of Proteus spp . (p < 0.05) and increased frequency of Pasteurella (p < 0.05) . The alpha-haemolytic streptococci were less common in T2 and TF2 than in other groups (NS) . Antibiotic treatment of cows before placental shedding (T2 or TF2, n = 6) postponed detachment of placenta compared to cows were no antibiotics were administered before placental shedding (T1, TF1, F1, F2 and 0, n = 16 . 9.8 days pp (median) versus p = 0.004) . Neither treatment shortened uterine involution . Flunixin treatments did not seem to influence recovery from infection or uterine involution . It was concluded that early oxytetracycline treatment of retained fetal membranes in the cow did not shorten the uterine involution or uterine infection but it did slow down the detachment process of the retained placenta . Oxytetracycline treatment after placental shedding might shorten the uterine infection but otherwise did not affect the clinical results . Flunixin treatment had no influence on the clinical outcome of the disease. Contemp Top Lab Anim Sci, 2002 Jan, 41(1), 43 - 5 Otitis interna as a result of Pasteurella multocida infection in a laboratory woodchuck; McCrory WP et al.; A laboratory woodchuck presented clinically with left-sided torticollis and a purulent exudate within the external auditory meatus of the left ear . Bacterial culture of the exudate resulted in a heavy growth of Pasteurella multocida . Treatment was initiated with topical and systemic antimicrobial compounds . There was no clinical improvement after 72 h of treatment, and euthanasia was elected . Radiographs correlated well with necropsy findings, confirming a diagnosis of otitis media; otitis interna was not confirmed but was suggested by the clinical presentation . To the authors knowledge, this is the first description of otitis media/interna as a result of P . multocida infection in a laboratory woodchuck. Antimicrob Agents Chemother, 2002 Mar, 46(3), 866 - 70 In vitro activities of the des-fluoro(6) Quinolone BMS-284756 against aerobic and anaerobic pathogens isolated from skin and soft tissue animal and human bite wound infections; Goldstein EJ et al.; BMS-284756, a new des-fluoro(6) quinolone, was very active against 240 aerobic and 180 anaerobic isolates from bite victims . It inhibited 403 of 420 (96%) isolates, including those of Moraxella spp., CDC group EF-4, and Eikenella corrodens at < or = 2 microg/ml and those of all Pasteurella spp . and Bergeyella zoohelcum at < or = 0.015 microg/ml . Fusobacterium russii and 6 of 11 Fusobacterium nucleatum isolates of animal bite origin were resistant, but isolates of human bite origin were susceptible, which suggests that they were of a different subspecies. Vet Rec, 2002 Jan 26, 150(4), 109 - 14 Comparative evaluation of ultrasonography and lung function testing with the clinical signs and pathology of calves inoculated experimentally with Pasteurella multocida; Reinhold P et al.; Seventeen calves were inoculated intratracheally with Pasteurella multocida 0 on three consecutive days with 10 ml of an inoculum containing 10(9) colony forming units/ml per day per calf . Before the first inoculation and 24 hours after the third, each calf was examined non-invasively by means of a clinical examination, chest ultrasonography, and impulse oscillometry to measure the impedance of the respiratory system . The inoculation of P multocida caused fever and a significant increase in respiratory rate and a decrease in tidal volume . There were also significant changes in the ultrasonographic results and in the impedance of the respiratory system . The percentage of the total surface area of the lungs showing pathological changes when the calves were euthanased 48 hours after the third inoculation ranged from 0.4 to 39 per cent . There were statistically significant correlations between the ultrasound scores and the pathological findings and between the ultrasound scores and the respiratory rate and tidal volume . The changes in the impedance of the respiratory system were not correlated with either the ultrasonographic or the pathological findings. J Clin Microbiol, 2002 Feb, 40(2), 687 - 9 Septicemia due to Pasteurella pneumotropica: 16S rRNA sequencing for diagnosis confirmation; Frebourg NB et al.; Bacteremia due to Pasteurella pneumotropica occurs infrequently . We report a case of septicemia in a 72-year-old woman who had no underlying illness . The microorganism was isolated from 10 blood cultures and identified by conventional and molecular methods . This is the first reported case of P . pneumotropica septicemia in an immunocompetent patient . The history of P . pneumotropica diseases in animals and humans and their varied clinical features are reviewed. J Clin Microbiol, 2002 Feb, 40(2), 666 - 8 Use of 16S rRNA sequencing for identification of Actinobacillus ureae isolated from a cerebrospinal fluid sample; Whitelaw AC et al.; Actinobacillus ureae, previously Pasteurella ureae, has on rare occasions been described as a cause of human infection . Owing to its rarity, it may not be easily identified in clinical microbiology laboratories by standard tests . This report describes a patient with acute bacterial meningitis due to A . ureae . The identity of the isolate was determined by means of DNA sequence analysis of a portion of the 16S rRNA gene. J Clin Microbiol, 2002 Feb, 40(2), 588 - 93 Typing of Pasteurella multocida isolated from pigs with and without porcine dermatitis and nephropathy syndrome; Lainson FA et al.; Porcine dermatitis and nephropathy syndrome (PDNS) is a sporadic, usually fatal disease of growing and finishing pigs that has been recognized in many pig-producing countries . Pasteurella multocida strains isolated from 15 pigs with PDNS and 51 pigs without PDNS were characterized by capsule and somatic antigen typing, random amplified polymorphic DNA (RAP-D) typing, and restriction analysis of genomic DNA using pulsed-field gel electrophoresis (PFGE) . While capsular, somatic, and RAP-D typing did not discriminate PDNS isolates from non-PDNS isolates, all of the isolates from PDNS cases showed an identical ApaI PFGE restriction pattern . This pattern was also found in a high proportion (36%) of P . multocida strains isolated from non-PDNS cases . Isolation of a single variant of P . multocida from tissues of pigs with PDNS warrants further investigation into the possible role of these bacteria in the etiology of the disease. J Clin Microbiol, 2002 Feb, 40(2), 461 - 5 Tiamulin activity against fastidious and nonfastidious veterinary and human bacterial isolates: initial development of in vitro susceptibility test methods; Jones RN et al.; Tiamulin is a pleuromutilin derivative used in veterinary practice for the control and specific therapy of infections in swine . This report summarizes studies to establish standardized susceptibility testing methods, interpretive criteria, and reagent details for use in veterinary methods recently developed by the National Committee for Clinical Laboratory Standards (NCCLS) (standards M31-A and M37-A, NCCLS, Wayne, Pa., 1999) . A total of 636 fastidious and nonfastidious animal and human pathogens were processed by using media and procedures described by the NCCLS . Tiamulin disk diffusion tests used a 30-microg disk concentration, and the proposed MIC breakpoints corresponding to levels achievable in animal target tissues (lung) were < or =4 microg/ml for susceptibility and > or =32 microg/ml for resistance . Correlate zone diameters for specific nonfastidious species were as follows: for Pasteurella multocida and staphylococci tested on Mueller-Hinton agar, susceptibility at > or =19 mm and resistance at < or =11 mm, and for Actinobacillus suis, Erysipelothrix rhusiopathiae, and Streptococcus suis tested on enriched chocolate Mueller-Hinton agar, susceptibility at > or =16 mm and resistance at < or =8 mm . When Actinobacillus pleuropneumoniae was tested, a susceptibility breakpoint of < or =16 microg/ml (> or =9 mm) was suggested for veterinary fastidious medium broth and enriched chocolate Mueller-Hinton agar . Absolute categorical agreement between NCCLS dilution and disk diffusion test results with these criteria ranged from 90.5 to 96.2% . Tiamulin susceptibility testing methods appear to be accurate in their categorical classification for indicated species, and their availability will allow immediate testing of animal isolates to guide therapy via appropriate levels of dosing and to monitor the development of resistance for agents in this unique class. Glycobiology, 2002 Jan, 12(1), 9R - 16R Microbial glycosaminoglycan glycosyltransferases; DeAngelis PL; Glycosaminoglycans, a class of linear polysaccharides composed of repeating disaccharide units containing a hexosamine, are important carbohydrates found in many organisms . Vertebrates utilize glycosaminoglycans in structural, recognition, adhesion, and signaling roles . Certain pathogenic bacteria produce extracellular capsules composed of glycosaminoglycans or glycosaminoglycan-like polymers that enhance the microbes' ability to infect or to colonize the host . In the period from 1993 to 2001, bacterial enzymes were discovered that catalyze the polymerization of the repeating unit of hyaluronan, chondroitin, or N-acetylheparosan (unsulfated, unepimerized heparin) . Depending on the specific carbohydrate and the microorganism, either a dual-action enzyme (synthase) that transfers two distinct monosaccharides or a pair of single-action transferases are utilized to synthesize the glycosaminoglycan polymer . Current views on the enzymology, structures, potential evolution, and the roles of the known glycosyltransferases from Streptococcus, Pasteurella, and Escherichia are discussed. Vet Immunol Immunopathol, 2002 Jan 1, 84(1-2), 97 - 110 Effect of experimental infection of cattle with bovine herpesvirus-1 (BHV-1) on the ex vivo interaction of bovine leukocytes with Mannheimia (Pasteurella) haemolytica leukotoxin; Leite F et al.; Mannheimia (Pasteurella) haemolytica A1 produces an extracellular leukotoxin (LKT) that is reported to bind the beta(2)-integrin CD11a/CD18 (LEA-1) on ruminant leukocytes . LKT binding induces activation, and subsequent cytolysis, of these cells . It is well known that active viral infection greatly increases the susceptibility of cattle to pasteurellosis . To better understand the mechanism by which this occurs, we investigated the effects of experimental in vivo infection of cattle with bovine herpes virus-1 (BHV-1) on the ex vivo interaction of bovine leukocytes with the M . haemolytica LKT . In this study, we demonstrated that active BHV-1 infection increased the expression of the beta(2)-integrin CD11a/CD18 (as defined by the mAb BAT75) on bovine peripheral blood neutrophils, enhanced the binding of LKT to bronchoalveolar lavage (BAL) leukocytes and peripheral blood neutrophils, and increased the killing of BAL leukocytes and peripheral blood leukocytes by LKT . In addition, BHV-1 greatly increased the number of BAL, resulting in many more LKT-responsive cells being present in the lungs . These findings might explain in part the increased susceptibility of BHV-1 infected cattle to pneumonic pasteurellosis. J Microbiol Immunol Infect, 2001 Dec, 34(4), 293 - 6 Pasteurella multocida bacteremia due to non-bite animal exposure in cirrhotic patients: report of two cases; Tseng HK et al.; Pasteurella species are very small gram-negative coccobacilli . They are normal flora found in the oral cavity and gastrointestinal tract of many animals, and can cause various infections including septicemia and pneumonia . Human infection with Pasteurella multocida occurs commonly as a localized cellulitis caused by animal bites . This report described 2 rare cases of P . multocida bacteremia in patients with liver cirrhosis and esophageal varices . Both patients had a history of contact with sick-appearing stray dogs, but neither had been bitten . P . multocida bacteremia should be included in the differential diagnosis of febrile cirrhotic patients with esophageal varices who have a history of non-bite animal exposure . Avoidance of animal contact by immunocompromised patients is the most important factor in preventing pasteurellosis. FEMS Microbiol Lett, 2002 Jan 2, 206(1), 25 - 30 Studies on the production of quorum-sensing signal molecules in Mannheimia haemolytica A1 and other Pasteurellaceae species; Malott RJ et al.; The bioluminescence assay system using Vibrio harveyi reporter strains were used to examine quorum-sensing autoinducer (AI) activity from Mannheimia haemolytica A1 cell-free culture supernatant . We showed that M . haemolytica A1 cell-free culture supernatant contains molecules that can stimulate the quorum-sensing system that regulates the expression of the luciferase operon in V . harveyi . Specifically, M . haemolytica A1 can stimulate only the quorum system 2 but not system 1, suggesting that the culture supernatant only contains molecules similar to AI-2 of V . harveyi . The bioluminescence assay was also used to show that culture supernatants from related Pasteurellaceae organisms, Pasteurella multocida, Pasteurella trehalosi, Actinobacillus suis and Actinobacillus pleuropneumoniae, also contain AI-2-like molecules . This is consistent with the presence of a luxS homolog in the genomes of P . multocida and A . pleuropneumoniae . A luxS homolog was cloned by PCR from M . haemolytica A1 using sequencing data from the ongoing genome sequencing project . The cloned luxS(M.h.) was able to complement AI-2 production in the Escherichia coli DH5alpha luxS mutant . This is the first report of a quorum-sensing activity in M . haemolytica A1 and suggests that this bacterium utilizes this mechanism to regulate expression of genes under specific conditions. Avian Dis, 2001 Oct-Dec, 45(4), 946 - 52 Ultrastructural location of the cross-protection factor on in vivo and in vitro grown Pasteurella multocida; Rimler RB et al.; Pasteurella multocida from infected turkey tissues expresses a unique immunogen called cross-protection factor (CPF) that induces immunity to challenge by both homologous and heterologous serotypes . In this study, we used a monoclonal antibody (AMP MAb) to CPF and protein A-colloidal gold (PACG) to locate CPF on P . multocida . After incubation with AMP MAb and PACG, CPF was detected at the bacterial surface and cell periphery of P . multocida in infected turkey liver and P . multocida isolated from infected turkey blood . CPF was not detected on P . multocida incubated with control monoclonal antibody . Pasteurella multocida isolated from infected turkey blood and cultivated in the peptone-based medium did not express CPF consistently, and some cells contained more CPF than others . The location of CPF also varied, and CPF was detected both intracellular and extracellular on the cell surface . In the latter cells, CPF was heavily concentrated to a specific lateral site or detected sloughing from the cell surface . These results correlate with laboratory observations that CPF detected on P . multocida from infected turkey tissues, P . multocida isolated from infected turkey blood, and P . multocida cultivated in peptone-based medium is associated with outer membrane fractions. Avian Dis, 2001 Oct-Dec, 45(4), 807 - 12 DNA fingerprint patterns of Pasteurella multocida from the same turkey farm on the same and different years; Olson LD et al.; The DNA fingerprint profiles of 126 isolants of Pasteurella multocida from 41 turkey farms in Missouri were analyzed after digestion with the restriction endonuclease HhaI and compared with their somatic antigenic type . The goal was to determine if the same isolant of P . multocida was reisolated from the the same farm during the same and consecutive years and after an interval of one or more years . Of the 37 pairs of P . multocida collected during the same year from the same turkey farms, the DNA fingerprint profiles were the same with 26 pairs (70.3%) and different with 11 pairs (29.7%) . Of the 33 pairs of P . multocida collected during consecutive years from the same 22 turkey farms, 21 pairs (63.6%) were the same and 12 pairs (36.4%) were different . Of the 15 pairs of P . multocida collected with an interval of one or more years between them from the same 14 turkey farms, only four pairs (26.7%) were the same and 11 pairs (73.3%) were different . There did not appear to be any relationship between the DNA fingerprint profiles and the typing of their somatic antigens because, although 44 pairs of isolants had the same DNA fingerprint profile and somatic antigenic type, 42 pairs differed in these parameters when all pairs were combined. Clin Diagn Lab Immunol, 2002 Jan, 9(1), 28 - 32 Increased anionic peptide distribution and intensity during progression and resolution of bacterial pneumonia; Fales-Williams AJ et al.; Anionic peptides (APs) are small anionic antimicrobial peptides composed of 7 aspartic acid residues and are produced in the lungs of humans, sheep, and cattle . Although expression by epithelial cells of some antimicrobial peptides (e.g., beta-defensins) of humans and ruminants is increased in response to acute infection, AP expression is not increased during acute infection, which suggests that the expression of the latter peptide is constitutive . In this study, the degree of AP expression during the progression (acute, subacute, and chronic) of bronchopneumonia was determined . Mannheimia (Pasteurella) haemolytica, a known inducer of bovinebeta-defensins, was inoculated intrabronchially with a fiber-optic bronchoscope in nine 3-month-old sheep, and tissues were collected at 1, 15, and 45 days postinoculation (p.i.); nine control animals received pyrogen-free saline by the same procedure and were killed at the same time points . In the acute group (1 day p.i.), the lungs had lesions typical of bronchopneumonia and the distribution and intensity of AP immunoreactivity (AP-IR) were similar to those of previous studies (minimal intensity and distribution of AP-IR in bronchiolar epithelial cells) . In the subacute group (15 days p.i.), there was prominent hyperplasia of bronchiolar and alveolar epithelial cells, and the chronic group (45 days p.i.) had yet more pronounced hyperplasia . In the subacute and chronic groups, the intensity and distribution of AP-IR in the cytoplasm of hyperplastic bronchiolar and type II alveolar cells were significantly increased compared to those of saline-inoculated and contralateral (noninoculated) lung lobes . Although AP expression appears constitutive, the constitutive production of AP is higher in hyperplastic, less differentiated cells than in fully differentiated, mature cells of the respiratory airways . The increased intensity and distribution of AP-IR in immature (hyperplastic) epithelial cells may be a mechanism by which production of a noninducible antimicrobial is increased temporarily during lesion progression and repair . This increased production of AP by hyperplastic cells may protect the lung against further infection until new, fully differentiated epithelial cells are capable of expressing their own inducible array of antimicrobial peptides. Vet Parasitol, 2001 Dec 13, 102(3), 225 - 34 The influence of T . evansi infection on the immuno-responsiveness of experimentally infected water buffaloes; Holland WG et al.; In order to define the immuno-suppressive capacity of Trypanosoma evansi infections in buffaloes on the induction of immune responses against heterologous antigens, infected and non-infected buffaloes were vaccinated against Pasteurella multocida (haemorrhagic septicemia) and were simultaneously immunised with a control antigen, human serum albumin (HSA) . Antibody responses against HSA were significantly reduced in T . evansi-infected animals, but no conclusive data were obtained on the antibody responses against P . multocida . Conversely, the local inflammatory response at the site of Pasteurella vaccination, as measured by increase in size, was significantly reduced in T . evansi-infected animals.These results indicate that the inductive capacity to mount humoral and cell-mediated immune responses against heterologous antigens is suppressed in T . evansi-infected animals . Consequently, T . evansi infection might interfere with the development of protective immunity upon heterologous vaccinations and could explain the poor protection of Pasteurella-vaccinated buffaloes in T . evansi-endemic areas of Vietnam. Can J Vet Res, 2001 Oct, 65(4), 213 - 22 Treatment of pigs experimentally infected with Mycoplasma hyopneumoniae, Pasteurella multocida, and Actinobacillus pleuropneumoniae with various antibiotics; Stipkovits L et al.; The authors have performed a comparative study of the efficacy of various in-feed medications for the treatment of 5- to 6-week-old specific pathogen-free (SPF) piglets experimentally infected on day 1 with Mycoplasma hyopneumoniae, on day 8 with Pasteurella multocida (serotype A), and on day 15 with Actinobacillus pleuropneumoniae (serotype 2) . The treatment started on day 9 and continued for 12 consecutive days, then the piglets were euthanized for examination of macroscopic, histologic, and pathologic lesions and for the presence of mycoplasmas and bacteria in the lungs . Based on the results of clinical observations (respiratory signs, rectal temperature, body weight gain, and feed conversion efficiency), macroscopic and histologic lesions of the lungs, and microbiologic findings, the best results were obtained by treatment of pigs with Econor + chlortetracycline, followed by Tetramutin, Pulmotil, Cyfac, and lincomycin + chlortetracycline. J Am Vet Med Assoc, 2001 Dec 15, 219(12), 1739 - 42 Health and performance of young dairy calves vaccinated with a modified-live Mannheimia haemolytica and Pasteurella multocida vaccine; Aubry P et al.; OBJECTIVE: To evaluate the health and performance of young dairy calves vaccinated with a commercial Mannheimia haemolytica and Pasteurella multocida vaccine . DESIGN: Randomized clinical trial . ANIMALS: 358 Holstein dairy calves between 14 and 20 days of age on 8 farms . PROCEDURE: Calves were randomly assigned to a control or vaccinated group . The vaccine used was a commercial modified-live M . haemolytica and P . multocida vaccine that was administered on days 0 and 14 . Calf weight was measured on day 0 and monthly for 3 months . Farmers were asked to record any treatment given to the calves and the reason for treatment during the 4 months of the study . Blood was collected from all calves on days 0 and 28, and titers of antibodies to M . haemolytica were determined by means of direct bacterial agglutination . RESULTS: Mean daily gain was not significantly different between vaccinated and control calves . Vaccinated calves had a significantly greater increase in antibody titers (5.3-fold increase), compared with control calves (3.6-fold increase) . There was no significant difference between vaccinated and control calves for any of the treatment outcomes (number and duration of treatments and age at first and last treatments) . CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that the M . haemolytica and P . multocida vaccine, given twice 2 weeks apart, was effective in increasing titers of antibodies against M . haemolytica in young dairy calves but did not improve calf performance or health. J Biol Chem, 2002 Mar 1, 277(9), 7209 - 13 Epub 2001 Dec 26. Identification and molecular cloning of a heparosan synthase from Pasteurella multocida type D; DeAngelis PL et al.; Pasteurella multocida Type D, a causative agent of atrophic rhinitis in swine and pasteurellosis in other domestic animals, produces an extracellular polysaccharide capsule that is a putative virulence factor . It was reported previously that the capsule was removed by treating microbes with heparin lyase III . We molecularly cloned a 617-residue enzyme, pmHS, which is a heparosan (nonsulfated, unepimerized heparin) synthase . Recombinant Escherichia coli-derived pmHS catalyzes the polymerization of the monosaccharides from UDP-GlcNAc and UDP-GlcUA . Other structurally related sugar nucleotides did not substitute . Synthase activity was stimulated about 7-25-fold by the addition of an exogenous polymer acceptor . Molecules composed of approximately 500-3,000 sugar residues were produced in vitro . The polysaccharide was sensitive to the action of heparin lyase III but resistant to hyaluronan lyase . The sequence of the pmHS enzyme is not very similar to the vertebrate heparin/heparan sulfate glycosyltransferases, EXT1 and 2, or to other Pasteurella glycosaminoglycan synthases that produce hyaluronan or chondroitin . The pmHS enzyme is the first microbial dual-action glycosyltransferase to be described that forms a polysaccharide composed of beta4GlcUA-alpha4GlcNAc disaccharide repeats . In contrast, heparosan biosynthesis in E . coli K5 requires at least two separate polypeptides, KfiA and KfiC, to catalyze the same polymerization reaction. FEMS Microbiol Lett, 2001 Dec 18, 205(2), 283 - 90 Occurrence and linkage of genes coding for resistance to sulfonamides, streptomycin and chloramphenicol in bacteria of the genera Pasteurella and Mannheimia; Kehrenberg C et al.; Twenty-three isolates of the two genera Pasteurella (P.) and Mannheimia (M.) were analysed for the presence of genes specifying resistance to sulfonamides, streptomycin, and chloramphenicol . Specific PCR assays for the detection of the genes sulII, strA and catAIII, but also for the confirmation of their physical linkage were developed . A resistance gene cluster consisting of all three genes and characterised by a PCR amplicon of 2.2 kb was detected on four different types of plasmids and also in the chromosomal DNA of seven isolates . Physically linked sulII and strA genes were detected on three different types of plasmids and in the chromosomal DNA of three isolates . Sequence analysis of the different PCR amplicons revealed that these genes were present in either the orientation sulII-strA separated by differently sized spacer sequences, or strA-sulII . A truncated strA gene preceding a sulII gene was also detected in two cases. Vet Microbiol, 2002 Feb 4, 84(4), 337 - 56 Leukotoxins of gram-negative bacteria; Narayanan SK et al.; Leukotoxins are a group of exotoxins that produce their primary toxic effects against leukocytes, especially polymorphonuclear cells (PMNs) . Leukotoxins include a variety of chemicals ranging from 9,10-epoxy 12-octadecenoate, a fatty acid derivative secreted by leukocytes themselves, to proteins such as RTX (repeats in toxin) . This review focuses on leukotoxins of three species of gram-negative bacteria, Mannheimia (Pasteurella) haemolytica, Actinobacillus actinomycetemcomitans, and Fusobacterium necrophorum. J Bacteriol, 2002 Jan, 184(1), 266 - 77 Mosaic structure and molecular evolution of the leukotoxin operon (lktCABD) in Mannheimia (Pasteurella) haemolytica, Mannheimia glucosida, and Pasteurella trehalosi; Davies RL et al.; The mosaic structure and molecular evolution of the leukotoxin operon (lktCABD) was investigated by nucleotide sequence comparison of the lktC, lktB, and lktD genes in 23 Mannheimia (Pasteurella) haemolytica, 6 Mannheimia glucosida, and 4 Pasteurella trehalosi strains . Sequence variation in the lktA gene has been described previously (R . L . Davies et al., J . Bacteriol . 183:1394-1404, 2001) . The leukotoxin operon of M . haemolytica has a complex mosaic structure and has been derived by extensive inter- and intraspecies horizontal DNA transfer and intragenic recombination events . However, the pattern of recombination varies throughout the operon and among the different evolutionary lineages of M . haemolytica . The lktA and lktB genes have the most complex mosaic structures with segments derived from up to four different sources, including M . glucosida and P . trehalosi . In contrast, the lktD gene is highly conserved in M . haemolytica . The lktC, lktA, and lktB genes of strains representing the major ovine lineages contain recombinant segments derived from bovine or bovine-like serotype A2 strains . These findings support the previous conclusion that host switching of bovine A2 strains from cattle to sheep has played a major role in the evolution of the leukotoxin operon in ovine strains of M . haemolytica . Homologous segments of donor and recipient alleles are identical, or nearly identical, indicating that the recombinational exchanges occurred relatively recent in evolutionary terms . The 5' and 3' ends of the operon are highly conserved in M . haemolytica, which suggests that multiple horizontal exchanges of the complete operon have occurred by a common mechanism such as transduction . Although the lktA and lktB genes both have complex mosaic structures and high nucleotide substitution rates, the amino acid diversity of LktB is significantly lower than that of LktA due to a higher degree of evolutionary constraint against amino acid replacement . The recombinational exchanges within the leukotoxin operon have had greatest effect on LktA and probably provide an adaptive advantage against the host antibody response by generating novel antigenic variation at surface-exposed sites. FEBS Lett, 2001 Nov 30, 509(1), 41 - 6 N-Acetylneuraminic acid uptake in Pasteurella (Mannheimia) haemolytica A2 occurs by an inducible and specific transport system; Solana S et al.; The N-acetylneuraminic acid (NeuAc) transport system of Pasteurella (Mannheimia) haemolytica A2 was studied when this bacterium was grown in both complex and chemically defined media . Kinetic measurements were carried out at 37 degrees C in 50 mM Tris-HCl buffer, pH 8.0, containing 50 microg/ml bovine serum albumin . Under these conditions, the uptake rate was linear for at least 3 min and the calculated K(m) for NeuAc was 0.1 microM . The transport rate was increased by the addition of several cations and was inhibited by sodium arsenite (95%), N,N'-dicyclohexyl-carbodiimide (50%), and 2,4-dinitrophenol (40%) at final concentration of 1 mM (each) . These results support the notion that NeuAc uptake is an active sugar cation symporter . Study of specificities showed that glucosamine, mannose and mannosamine inhibited the transport of NeuAc in this bacterium . Analysis of expression revealed that the NeuAc transport system was induced by NeuAc and by the simultaneous presence of glucose and galactose in the growth medium. Vet Microbiol, 2002 Jan 3, 84(1-2), 169 - 77 Antimicrobial susceptibility and plasmid analysis of Actinobacillus pleuropneumoniae isolated in Taiwan; Chang CF et al.; Sixty Actinobacillus pleuropneumoniae (App) strains from pigs in Taiwan were examined . Serotyping revealed that these belonged to serovars 1 (n=53), 2 (n=3), and 5 (n=4) . Agar disk diffusion susceptibility testing of the isolates showed 55 (92%) were resistant to three or more antimicrobial agents . Six resistance patterns were observed . Ampicillin-chloramphenicol-flumequine-nalidixic acid-streptomycin-sulfonamide/trimethoprim-tetracycline was the most common multi-resistance pattern . Minimal inhibitory concentration of 14 antimicrobial agents was determined . The isolates were highly susceptible to ceftiofur and trimethoprim in vitro . Isolates were resistant to streptomycin, ampicillin, and nalidixic acid . All isolates were examined for the presence of plasmids using the alkaline lysis method . Forty three (72%) isolates had four plasmid bands with an approximate sizes of 3.5, 4.3, 5.8 and 6.0 kb; 12 (20%) had three bands at 3.5, 4.3 and 5.2 kb, and 5 (8%) had no plasmid bands . Antimicrobial resistance plasmids were detected in resistant strains of App . Three antimicrobial resistance plasmids were transformed into E . coli DH5 alpha . pTMY1 (4.3 kb) encoded a streptomycin kinase and a dihydropteroate synthase; pTMY2 (6.0 kb) encoded ROB-1 beta-lactamase and aminoglycoside 3'-phosphotransferase; pTMY3 (5.2 kb) encoded only ROB-1 beta-lactamase . The 4.3 kb plasmid was sequenced and consisted of 4242 bp with 42.9% GC content . The 4.3 kb plasmid DNA sequence was 98% homologous to a plasmid previously isolated from Pasteurella haemolytica. Vet Microbiol, 2002 Jan 3, 84(1-2), 103 - 14 Phenotypic and genotypic characterization of Mannheimia (Pasteurella) haemolytica-like strains isolated from diseased animals in Denmark; Angen O et al.; Trehalose-negative strains of the Pasteurella haemolytica complex have recently been transferred to a new genus, Mannheimia . This genus presently consists of five named species: M . haemolytica, M . glucosida, M . granulomatis, M . ruminalis and M . varigena . The purpose of this study was to investigate the occurrence of these species and lesions associated with these isolates in Denmark . In all 106 M . haemolytica-like strains isolated from pathological material from cattle, sheep, pigs and hares submitted to the Danish Veterinary Laboratory between 1994 and 1998 were investigated . Phenotypic characterization and ribotyping were used for identification in addition to sequencing of the 16S rRNA genes for selected strains . The species allocation was determined by comparison to results from a previous polyphasic taxonomic study . Seventy-one percent of the strains belonged to M . haemolytica, 18% to M . varigena and 8% to unnamed groups within the genus Mannheimia . Single isolates identified as M . glucosida and P . trehalosi, respectively, were detected . Two isolates belonged to M . granulomatis . Forty-three percent of the strains belonged to serotype 1, 41% were untypeable, while the rest belonged to serotypes 2, 7, 9, and 16 . The present investigation also showed that a simplified phenotypic characterization using Diatabs Diagnostic Tablets (Rosco, Denmark) represents a useful method for obtaining a quick and reliable species identification . Finally, the investigation confirmed that serotyping does not represent a reliable method for species identification . The heterogeneity of species associated with bovine "pasteurellosis" should be considered in future studies to improve our understanding of the pathogenesis of pneumonic disease. Vet Microbiol, 2002 Jan 3, 84(1-2), 69 - 78 Plasmid and restriction endonuclease patterns in Pasteurella multocida isolated from a swine pyramid; Rubies X et al.; Restriction endonuclease analysis (REA) and plasmid profile were used to study the epidemiology of Pasteurella multocida in a swine pyramid structure . The studied pyramid was comprised of a group of 12 swine farrow-to-finish farms related by unidirectional animal movement . P . multocida isolates were obtained from the lungs of 275 slaughtered pigs . Serotyping was performed by hyaluronidase sensitivity test and toxicity was investigated by the ELISA test . HpaII was used to cleave the P . multocida extracted DNA . REA patterns relationships were studied using the Sokal-Michener coefficients, and the dendrogram was built using the UPGMA system . The 218 P . multocida isolates obtained were distributed in 17 REA patterns . In 9 of the 12 farms studied only 2-3 REA patterns were detected, with one clearly predominant pattern . The 81 strains with plasmids were assigned to six plasmid profiles . REA and plasmid profiles proved to be good epidemiological tools for identifying different strains of P . multocida with the same phenotype. Vet Rec, 2001 Nov 10, 149(19), 583 - 7 Safety and efficacy of an oil-adjuvant vaccine against haemorrhagic septicaemia in buffalo calves: cross-protection between the serotypes B:2,5 and E:2,5; Shah NH et al.; The safety, efficacy and duration of immunity of an improved oil-adjuvant vaccine against haemorrhagic septicaemia, containing inactivated cells of Pasteurella multocida serotype B:2,5, were tested in young buffalo calves in Pakistan . For safety testing, five buffalo calves were vaccinated intramuscularly with twice the normal dose, and six weeks later with a normal dose . Except for a transient rise in rectal temperature at six hours after the vaccinations, no systemic reactions were observed . The buffaloes remained in good condition and had a normal appetite . No local reactions were observed at the injection site . For efficacy testing two trials were carried out . In the first, buffalo calves were vaccinated intramuscularly either with two doses two-and-a-half months apart, or with a single dose, or left unvaccinated . They were challenged subcutaneously with virulent P multocida after eight, 13 or 15 months . After challenge at eight months the four buffaloes given two doses and the buffalo given one dose were protected, whereas the control animal developed the typical signs of the disease . After the challenges at 13 and 15 months, the vaccinated animals were still protected whereas the control animals died . In the second trial, buffalo calves were vaccinated intramuscularly either with two doses two months apart, or with a single dose at two months or left unvaccinated . The buffaloes were challenged after eight or 14 months . After challenge at eight months the four control animals died, whereas three of the four buffaloes given a single dose were protected . After challenge at 14 months, the three control animals died, whereas four of the five buffaloes given two doses and both the buffaloes given a single dose were protected . To test for cross-protection against the heterologous serotypes E:2,5 and B:3,4, groups of mice were vaccinated once or left unvaccinated . Four weeks later, the vaccinated and control groups were challenged with a dilution series of the different challenge cultures . The vaccine appeared to induce protection against challenge with different strains of serotypes B:2,5 and E:2,5 but not against strains of serotype B:3,4. Vet Rec, 2001 Nov 3, 149(18), 549 - 52 Relationships between clinical and pathological signs of disease in calves infected with Mannheimia (Pasteurella) haemolytica type A1; Reeve-Johnson L; Respiratory disease was induced in 16 calves, and the terminal clinical signs of disease and postmortem pathological observations were recorded . By Spearman's correlation coefficient, the respiratory rate, rectal temperature and clinical scores of the calves were significantly correlated with the extent of lung consolidation . The respiratory rate was the clinical sign most consistently correlated with the other clinical and pathological signs of respiratory disease. Infect Immun, 2001 Dec, 69(12), 7839 - 50 Localization of functional domains of the mitogenic toxin of Pasteurella multocida; Pullinger GD et al.; The locations of the catalytic and receptor-binding domains of the Pasteurella multocida toxin (PMT) were investigated . N- and C-terminal fragments of PMT were cloned and expressed as fusion proteins with affinity tags . Purified fusion proteins were assessed in suitable assays for catalytic activity and cell-binding ability . A C-terminal fragment (amino acids 681 to 1285) was catalytically active . When microinjected into quiescent Swiss 3T3 cells, it induced changes in cell morphology typical of toxin-treated cells and stimulated DNA synthesis . An N-terminal fragment with a His tag at the C terminus (amino acids 1 to 506) competed with full-length toxin for binding to surface receptors and therefore contains the cell-binding domain . The inactive mutant containing a mutation near the C terminus (C1165S) also bound to cells in this assay . Polyclonal antibodies raised to the N-terminal PMT region bound efficiently to full-length native toxin, suggesting that the N terminus is surface located . Antibodies to the C terminus of PMT were microinjected into cells and inhibited the activity of toxin added subsequently to the medium, confirming that the C terminus contains the active site . Analysis of the PMT sequence predicted a putative transmembrane domain with predicted hydrophobic and amphipathic helices near the N terminus over the region of homology to the cytotoxic necrotizing factors . The C-terminal end of PMT was predicted to be a mixed alpha/beta domain, a structure commonly found in catalytic domains . Homology to proteins of known structure and threading calculations supported these assignments. Aust Vet J, 2001 Sep, 79(9), 634 - 9 Characterisation of a novel Mannheimia sp from Australian feedlot cattle; Blackalu P et al.; OBJECTIVE: To characterise eight isolates of a Gram-negative organism obtained from the upper respiratory tract of cattle showing evidence of mild upper respiratory tract disease . DESIGN: The isolates were compared with the five recognised species within the genus Mannheimia - M haemolytica, M glucosida, M granulomatis, M ruminalis and M varigena--using a range of phenotypic and genotypic methods . RESULTS: Phenotypic characterisation indicated that the isolates belonged to the trehalose-negative {Pasteurella} haemolytica complex . This complex has recently been reorganised into five species within the new genus Mannheimia . Ribotyping performed using HindIII and a computerised analysis system indicated that the eight Australian isolates formed a distinct cluster that was related to, but different from, the five recognised species of Mannheimia . The 16S rRNA sequence of one isolate (BNO311) was determined and a phylogenetic analysis performed . Isolate BNO311 was distinct from the five named Mannheimia spp but did join a larger cluster consisting of rRNA cluster IV (M varigena) and the unnamed rRNA cluster V of Mannheimia . DNA:DNA hybridisation between isolate BNO311 and M haemolytica NCTC 9380T, M granulomatis P411 and Actinobacillus ligniersii NCTC 4189T all suggested similarities of approximately 30% . CONCLUSIONS: These phenotypic and genotypic characterisation studies suggest that the eight Australian isolates represent a new species of Mannheimia . Until further characterisation studies are performed, we are unwilling to propose a name for this taxon, preferring to refer to this possible new species as Bisgaard taxon 39 of cluster V of Mannheimia. South Med J, 2001 Oct, 94(10), 1033 - 5 Prosthetic valve endocarditis caused by Pasteurella dagmatis; Rosenbach KA et al.; This case of prosthetic valve endocarditis due to Pasteurella dagmatis is the first to be reported in the English language medical literature . The two reported cases of native valve endocarditis due to P dagmatis are reviewed, and the treatment of Pasteurella-induced endocarditis is discussed. J Vet Pharmacol Ther, 2001 Oct, 24(5), 353 - 8 A field comparison of the efficacy and tolerance of marbofloxacin in the treatment of bovine respiratory disease; Thomas E et al.; A multicentre, controlled, randomized and blinded trial was carried out in 180 ruminating calves with pyrexia and respiratory sign(s) on nine Belgian, British and French farms . All animals were sampled for pathogenic bacteria before treatment and at failure/relapse . Calves were injected with either marbofloxacin (M) solution {Marbocyl (Laboratoire Vetoquinol, Lure, France) 10%} at 2 mg/kg/24 h for 4 days intravenously on the first day then subcutaneously, or tilmicosin (T) solution (Micotil, Elanco Products Ltd, Basingstoke, Hants, UK) at 10 mg/kg as a single subcutaneous (s.c.) injection . The animals were examined clinically eight times up to day 28 . The bacterial pathogens were found to be sensitive to marbofloxacin: for Pasteurella haemolytica the minimum inhibitory concentration (MIC)90 was 0.08 microg/mL and for P . multocida the MIC90 was 0.04 microg/mL . Cure rates at day 4 for group M and group T were 84 vs . 82%, respectively (P > or = 0.05) . However, overall clinical score was significantly lower after 1 day in group M (P < 0.05) . There was no difference in either relapse rate or average daily weight gain between groups . Marbofloxacin was found to be better tolerated than tilmicosin at the s.c . injection site (77.5 vs . 42.2% calves without local swelling, P=0.001) and was well tolerated when injected intravenously . Marbofloxacin was shown to have comparable but faster efficacy and better local tolerance than tilmicosin in the treatment of bovine respiratory disease (BRD). Pathol Biol (Paris), 2001 Oct, 49(8), 606 - 11 {Comparative study of the bacteriostatic and bactericidal activity of levofloxacin against Pasteurella strains isolated from man}; Heurtin C et al.; The MICs of seven quinolones, nalidixic acid, pefloxacin, ofloxacin, d-ofloxacin, ciprofloxacin, sparfloxacin and levofloxacin, were determined by agar dilution method comparatively to those of amoxycillin, cefpodoxime, doxycyclin and clarithromycin against 75 clinical isolates of Pasteurella multocida, P . dagmatis and P . canis . Time-kill method was performed for three selected P . multocida isolates . Fluoroquinolones were the most active agents . At concentration of 0.016 mg/L of sparfloxacin or levofloxacin the 75 isolates were inhibited . The MICs of levofloxacin and sparfloxacin showed that the activity of these molecules was two to four times higher than that of the other quinolones studied . Time-kill studies showed a complete killing in six hours with the CMI x 2 of pefloxacin, ofloxacin, ciprofloxacin, sparfloxacin and levofloxacin . This result was obtained more rapidly with the quinolones than with amoxicillin or cefpodoxime . Doxycycline and clarithromycin were devoid of bactericidal activity. Int J Med Microbiol, 2001 Sep, 291(4), 261 - 8 Pasteurella multocida toxin: the mitogenic toxin that stimulates signalling cascades to regulate growth and differentiation; Lax AJ et al.; Pasteurella multocida toxin (PMT) is an unusual toxin that acts as a mitogen by stimulating various intracellular signalling cascades . Pathways downstream of the G-protein Gq and also downstream of the Rho proteins are activated . Thus PMT action stimulates phospholipase C leading to activation of protein kinase C, an increase in inositol phosphates, and a rise in intracellular calcium . Rho activation of the Rho kinase leads to cytoskeletal reorganisation, tyrosine phosphorylation of the focal adhesion kinase, and activation of the Src proto-oncogene . In addition, signalling through the Ras-MAP kinase signalling pathway is also initiated . PMT is an intracellularly acting toxin, and functional domains that carry out different aspects of its function have been described . The intracellular target of the toxin is currently not known . PMT also acts to inhibit differentiation, in particular of bone cells, where it prevents the formation of mineralised bone nodules in vitro . The toxin is the causative agent of a porcine disease that is characterised by bone resorption . Injection of very low doses of toxin leads to proliferative effects, but at higher doses is lethal . The possible effect of PMT-induced perturbation of signal transduction pathways is discussed. J Antimicrob Chemother, 2001 Nov, 48(5), 631 - 40 Tetracycline resistance genes in isolates of Pasteurella multocida, Mannheimia haemolytica, Mannheimia glucosida and Mannheimia varigena from bovine and swine respiratory disease: intergeneric spread of the tet(H) plasmid pMHT1; Kehrenberg C et al.; Tetracycline-resistant isolates of Pasteurella multocida and Mannheimia spp . from respiratory diseases in cattle and swine were investigated for the classes of tet gene and their chromosomal or plasmid location . The 34 isolates comprised eight P . multocida, 23 Mannheimia haemolytica, two Mannheimia varigena and a single Mannheimia glucosida isolate . Identification of the tet genes was achieved by PCR analysis and hybridization with specific probes . Transformation and hybridization experiments served to confirm the plasmid location of tet genes . Selected tet genes and their adjacent regions were sequenced . The tet genes tet(B), tet(G) and tet(H) were detected . The gene tet(H) was present in 26 isolates . The 4.4 kb tet(H)-carrying plasmid pMHT1 was detected in six isolates representing all four species . In the remaining 28 isolates, copies of tet(B), tet(G) and tet(H) were identified as chromosomal . No correlation between the tet gene type and the MIC of tetracycline, or between the number of tet gene copies and the MIC of tetracycline was observed . Tetracycline resistance in P . multocida and Mannheimia spp . is mediated by at least three different tet genes . A new type of tet(H)- carrying plasmid, pMHT1, was identified . The detection of pMHT1 in M . glucosida and M . varigena is the first report of resistance plasmids in isolates of these two species . For the first time, tet(G) genes were detected in members of the family Pasteurellaceae. Vet Rec, 2001 Oct 6, 149(14), 412 - 7 Detection of Pasteurella multocida in pigs with porcine dermatitis and nephropathy syndrome; Thomson JR et al.; Comprehensive bacterial cultures were made on samples from 20 pigs that had died of porcine dermatitis and nephropathy syndrome after a short clinical illness . Eleven species of porcine bacterial pathogens and a range of commensal organisms were isolated . Pasteurella multocida was isolated from 16 of the 20 cases but the other pathogens occurred much less commonly . P . multocida was isolated from between one and five sites per case and from the tonsils, retropharyngeal lymph node or lungs in 14 of the 16 cases . Immunohistochemical investigations of kidneys from 30 cases of the syndrome (including the 20 cases in the bacteriological study) revealed P . multocida-specific staining in 26 of the cases, primarily in the renal tubular epithelial cells of the proximal convoluted tubules, but also in the glomeruli, in lesions of renal vasculitis and in the cytoplasm of interstitial mononuclear cells. Res Vet Sci, 2001 Jun, 70(3), 255 - 6 Cloning of 87 kDa outer membrane protein gene of Pasteurella multocida P52; Chaudhuri P et al.; Pasteurella multocida serotype B:2 is the causative agent of haemorrhagic septicaemia, a fatal disease of cattle and buffaloes . Formalin inactivated whole cell bacterin is used to prepare vaccines in India . However, outer membrane proteins (OMPS) of P . multocida were reported to be potential immunogens . The 87-kDa OMP of P . multocida P52, serotype B:2 has been identified as one of the major antigens because this protein reacted with serum of vaccinated animals . The gene omp87, encoding an 87-kDa OMP was amplified and cloned into pBluescript SK(-) vector . This gene was found to localise in a 9.0 kb Hind III fragment of P . multocida genome. Res Vet Sci, 2001 Jun, 70(3), 191 - 7 Ultrastructural pathology of nasal and tracheal mucosa of rabbits experimentally infected with Pasteurella multocida serotype D:1; Al-Haddawi MH et al.; Sixteen 8- to 9-week-old Pasteurella multocida-free New Zealand White rabbits were divided into two equal groups . The first group was inoculated intranasally with P multocida serotype D:1 strain and the second group that was inoculated with phosphate-buffered saline (PBS) only was used as a control group . Pasteurella multocida was isolated from the nasal cavity of all infected rabbits in group 1 and from tracheal swabs of seven rabbits in this group . Four rabbits in group 1 died with clinical signs of septicaemia, two rabbits had mucopurulent nasal discharge and pneumonic lesions and the other two did not show any clinical signs or gross lesions . The ultrastructural changes detected were deciliation or clumping of cilia of ciliated epithelium, cellular swelling, vacuolation and sloughing . The subepithelial capillaries showed congestion, intravascular fibrin deposition, platelets aggregation and endothelial injury . Pasteurella multocida was observed attached to the injured endothelial cells . Heterophils, mast cells, vacuolated monocytes and macrophages infiltrated the lamina propria and between the degenerated epithelial cells. J Vet Med B Infect Dis Vet Public Health, 2001 Sep, 48(7), 555 - 60 Antimicrobial susceptibility of Pasteurella multocida isolated from cattle and pigs; Yoshimura H et al.; Minimum inhibitory concentrations (MICs) of 10 antimicrobial agents were determined for Pasteurella multocida from cattle and pigs (72 and 68 isolates, respectively) . Higher MICs were observed with oxytetracycline, doxycycline, tilmicosin and thiamphenicol for porcine isolates than for bovine isolates . Enrofloxacin was the most active, with an MIC for 90% of the isolates (MIC90) of 0.05 microg/ml for both bovine and porcine isolates . Aspoxicillin exhibited the same excellent activity against penicillin-susceptible isolates as ceftiofur, with MICs ranging from < or = 0.025 to 0.1 microg/ml . Aminoglycosides were less active, with an MIC90 of > 100 microg/ml for both bovine and porcine isolates. J Vet Med B Infect Dis Vet Public Health, 2001 Sep, 48(7), 513 - 8 The effect of Pasteurella haemolytica A2 infection on phagocytosis efficiency of caprine broncho-alveolar macrophages; Zamri-Saad M et al.; An experiment was designed to study the in vivo effect of Pasteurella haemolytica A2 infection on the phagocytosis activity of caprine broncho-alveolar macrophages and the extent of pneumonic lesions . Twelve healthy local Kacang goats, about 7 months of age, were divided into two groups of six . Goats in group 1 were inoculated intratracheally with 4 ml inoculum containing 2.8 x 10(9) colony-forming units (CFU)/ml of Staphylococcus aureus . Goats in group 2 were inoculated intratracheally with 4 ml of inoculum containing 9.5 x 10(8) CFU/ml of Pasteurella haemolytica A2 isolated earlier from pneumonic lungs of goat . At intervals of 3 and 7 days post-challenge five goats from each group were killed and the lungs were washed with sterile phosphate-buffered saline . Smears were prepared from the lung washing fluid and the number of macrophages with phagocytic activity was determined . At day 3 post-infection, goats of both groups showed a similar pattern of pneumonic lesion . The lung washing fluid of goats in group 2 was found to contain numerous neutrophils and macrophages . Goats in group 2 showed significantly (P < 0.05) higher extent of lung lesions than group 1 . Similarly, the average extent of lung lesions was significantly (P < 0.05) more severe in group 2 at day 7 post-infection . The lung washing fluid contained mostly macrophages . The phagocytic activity following S . aureus infection was more efficient and significantly (P < 0.01) higher compared with infection by P . haemolytica A2 . There were weak correlations between the extent of pneumonic lesion and the phagocytic activity . Thus, goats with poor phagocytic activity were likely to develop more extensive lung lesions. J Vet Med Sci, 2001 Sep, 63(9), 1055 - 6 Isolation of Pasteurella multocida during an outbreak of infectious septicemia in Japanese quail (Coturnix coturnix japonica); Goto Y et al.; In May 1994, about fifty Japanese quails out of ninety being bred for experimental purposes at Miyazaki University died of acute septicemia within a few days . At autopsy, there were no gross pathological lesions, however, severe bacteremia was observed in all cases . Bacterial examination revealed the presence of Pasteurella multocida in blood and several organs in pure culture and they were of Carter's capsular type A, Heddleston's type 3-4 and Namioka's type O-8-9 . The LD50 of bacteria in quails and mice were 4.3 x 10(4) cfu and 3.9 x 10(2) cfu, respectively . All of the three chickens experimentally infected with 4 x 10(4) of the isolate died within 20 hr after the infection and several bacteria were recovered from their blood and organs . This, to our knowledge, is the first report on an outbreak of fowl cholera in Japanese quails in Japan. Kansenshogaku Zasshi, 2001 Sep, 75(9), 780 - 4 {A clinical study on patients detected Pasteurella multocida from sputum}; Gonda H et al.; We reported ten cases, (four female and six male), whose sputum cultures positive for Pasteurella multocida from 1990 to 2000 . In the past eleven years increasing numbers of cases have appeared in our hospital . The majority of the cases with P . multocida possessed some underlying pulmonary diseases (seven cases, 70%), inactive lung tuberculosis or bronchiectasis . There were compromised hosts such as high ages person, steroids dependent person and diabetes mellitus patients . P . multocida was almost susceptible to antibioticus (penicillin and cephalosporins), although some erythromycin resistant strains were identified . The cats' oral cavities in our two cases were cultured and P . multocida were isolated . In our survey the prevalence of this organism is as high as 85% in cats . Our data suggests that patients who are in the high infection risk category are easily infected to P . multocida. Bioorg Med Chem Lett, 2001 Oct 22, 11(20), 2751 - 4 Synthesis and structure-activity relationships of thiotetronic acid analogues of thiolactomycin; Sakya SM et al.; 3-Acetyl analogues of thiolactomycin, a thiotetronic acid natural product, were synthesized and profiled against livestock pathogens . Some analogues showed improved activity over thiolactomycin against Staphylococcus aureus and comparable activity against Pasteurella multocida . Several semisynthetically modified analogues of thiolactomycin showed no improvement in activity over thiolactomycin. Biochem J, 2001 Sep 15, 358(Pt 3), 585 - 98 Kinetic properties of the acylneuraminate cytidylyltransferase from Pasteurella haemolytica A2; Bravo IG et al.; Neuroinvasive and septicaemia-causing pathogens often display a polysialic acid capsule that is involved in invasive behaviour . N-Acetylneuraminic acid (NeuAc) is the basic monomer of polysialic acid . The activated form, CMP-Neu5Ac, is synthesized by the acylneuraminate cytidylyltransferase (ACT; EC 2.7.7.43) . We have purified this enzyme from Pasteurella haemolytica A2 to apparent homogeneity (522-fold) . The protein behaved homogeneously on SDS/PAGE as a 43 kDa band, a size similar to that of Escherichia coli, calf, mouse and rat . Specific activity in crude lysate displayed one of the highest values cited in the literature (153 m-units/mg) . We have studied the steady-state kinetic mechanism of the enzyme by using normalized plot premises . The catalysis proceeds through a Ping Pong Bi Bi mechanism, with CTP as the first substrate and CMP-NeuAc as the last product . The true Km values were 1.77 mM for CTP and 1.82 mM for NeuAc . The nucleotides CDP, UTP, UDP and TTP, and the modified sialic acid N-glycolylneuraminic acid were also substrates of the ACT activity . The enzyme is inhibited by cytidine nucleotides through binding to a second cytidyl-binding site . This inhibition is greater with nucleotides that display a long phosphate tail, and the genuine inhibitor is the substrate CTP . At physiological concentrations, ATP is an activator, and AMP an inhibitor, of the ACT activity . The activated sugar UDP-N-acetylglucosamine acts as an inhibitor, thus suggesting cross-regulation of the peptidoglycan and polysialic acid pathways . Our findings provide new mechanistic insights into the nature of sialic acid activation and suggest new targets for the approach to the pathogenesis of encapsulated bacteria. Microbiology, 2001 Oct, 147(Pt 10), 2739 - 48 Genetic diversity of Pasteurella multocida fowl cholera isolates as demonstrated by ribotyping and 16S rRNA and partial atpD sequence comparisons; Petersen KD et al.; The genetic diversity of Pasteurella multocida, the aetiological agent of fowl cholera, was investigated . The strain collection comprised 69 clinical isolates representing a wide spectrum of hosts and geographic origin . The three type strains for the subspecies of P . multocida were also included . Avian isolates of P . multocida subsp . multocida and P . multocida subsp . septica did not represent separate lines by HpaII ribotyping and the two type strains of mammalian origin (porcine and cat bite) seemed to be representative of avian strains of P . multocida subspp . multocida and septica . By ribotyping, all P . multocida subsp . gallicida strains, except one chicken isolate and the type strain, clustered together . This indicated that the bovine type strain was not representative of this subspecies and that most strains of P . multocida subsp . gallicida are genetically related and may be distantly related to other P . multocida isolates, including those of avian origin . By 16S rRNA and atpD sequence comparisons of selected strains, including both P . multocida isolated from birds and mammals and selected distantly related Pasteurella species associated with birds and mammals, it was found that P . multocida is monophyletic . Extended DNA-DNA hybridizations are highly indicated since strains may exist which would connect the existing subspecies at species level . The considerable genetic diversity of P . multocida fowl cholera isolates is probably related to the clonal nature of this organism, resulting in many divergent lines. Avian Dis, 2001 Jul-Sep, 45(3), 655 - 8 Pathogenicity and drug susceptibility of the Pasteurella anatis isolated in chickens in Taiwan; Lin MY et al.; A strain of Pasteurella anatis (PA) was isolated from the sinus of an adult leghorn laying chicken with sinusitis, nasal discharge, drop in egg production, and low mortality, symptoms initially thought to indicate infectious coryza . The tiny, smooth, whitish colonies were identified as PA . To compare its pathogenicity with that of commercial broilers, nine groups, 10 birds per group, of 10-day-old broilers were individually inoculated with the strain of PA, Pasteurella multocida (PM), or Escherichia coli (EC) by intravenous, intraperitoneal, intramuscular, or subcutaneous inoculation . The PA was determined to cause the signs, lesions, and septicemic death, which are similar to the symptoms of PM or EC infection . At 1 wk postinfection (PI), the mortality rate was between that of PM and EC infection at 1 wk PI . Twenty antimicrobial-containing discs were evaluated, and the isolate was highly sensitive to cetiofer, amoxicillin, lincopectin, and furazolidone . Furthermore, it was moderately sensitive to tetracycline and enrofloxacin and only slightly sensitive to cephalothin, chloramphenicol, flumequine, nalidixic acid, neomycin, oxolinic acid, streptomycin, and trimethoprim . The PA infection was treated successfully with amoxicillin. Avian Dis, 2001 Jul-Sep, 45(3), 572 - 80 Purification of a cross-protective antigen from Pasteurella multocida grown in vitro and in vivo; Rimler RB; A peptone-based medium was formulated to grow Pasteurella multocida in vitro, which expressed an antigen that induces cross protection in turkeys against different serotypes . Vaccines of various chromatographic fractions obtained from P . multocida grown in the medium induced active immune cross protection in turkeys, and sera from these turkeys passively cross protected naive poults . An antigen of approximately 39 kD molecular size was purified by preparative sodium dodecyl sulfate-polyacrylamide gel electrophoresis and electroelution from hydroxyapatite chromatographic fractions of both in vivo- and in vitro-grown P . multocida . The purified antigen from either source induced active immune cross protection but no passive protection in one of two experiments . Increasing the dose of vaccine resulted in both active and passive immune cross protection in the second experiment. Lett Appl Microbiol, 2001 Sep, 33(3), 216 - 21 Development of a 23S rRNA-based PCR assay for the identification of Pasteurella multocida; Miflin JK et al.; AIMS: The aim of this work was to develop a rapid diagnostic test for Pasteurella multocida . METHODS AND RESULTS: A polymerase chain reaction (PCR) assay using primers derived from the 23S rRNA gene sequence of Past . multocida was developed . The PCR assay correctly identified all 144 isolates of Past . multocida tested, including type strains of the three subspecies as well as the reference strains for the Heddleston and Carter typing schemes . Of 20 closely related bacteria from the family Pasteurellaceae tested, only the type strains of Past . canis biovar 2 and Past . avium biovar 2 were positive . These two bacteria, formerly known as Bisgaard Taxon 13, are the closest phylogenetic relatives of Past . multocida based on 16S ribosomal rRNA . All phylogenetically unrelated avian and porcine organisms tested were negative . CONCLUSION: This PCR enables rapid identification of Past . multocida colonies from avian or porcine origin . SIGNIFICANCE AND IMPACT OF THE STUDY: Veterinary diagnostic laboratories can use this PCR to rapidly and accurately diagnose fowl cholera and porcine pasteurellosis. Vaccine, 2001 Sep 14, 19(32), 4842 - 50 Bordetella bronchiseptica fimbrial protein-enhanced immunogenicity of a Mannheimia haemolytica leukotoxin fragment; Rajeev S et al.; Leukotoxin produced by Mannheimia (Pasteurella) haemolytica is an important virulence factor in shipping fever pneumonia in feedlot cattle and is a critical protective antigen . In this study, the immune response to a chimeric protein generated by combining a gene fragment encoding neutralizing epitopes of M . haemolytica leukotoxin and a fimbrial protein gene (fim N) from Bordetella bronchiseptica was evaluated . The recombinant gene was cloned in a bacterial expression vector under the control of the tac promoter and expressed as a fusion protein with glutathione-S-transferase (GST) in Escherichia coli . Immunization of mice with the recombinant protein, GST-LTXFIM elicited a significantly stronger anti-leukotoxin antibody response than comparable immunizations with GST-LTX fusion proteins lacking FIM N . The GST-LTXFIM was also more stable than GST-LTX during storage at -80 degrees C, thus alleviating a stability problem inherent to leukotoxin . This chimeric protein may be a candidate for inclusion in new generation vaccines against shipping fever pneumonia. Plasmid, 2001 Jul, 46(1), 60 - 4 A cryptic plasmid from Pasteurella multocida has a predicted protein nearly identical to a transport protein from Actinobacillus actinomycetemcomitans; McGee JE et al.; Several plasmids from Pasteurella multocida have been shown to carry antibiotic resistance genes but no other genes possibly related to the organism's pathogenesis . We report here that sequence from the plasmid pLEM from a fowl isolate of P . multocida, strain 1059, contained one open reading frame that had significant identity with a predicted protein from pVT745, a plasmid that was isolated from a human oral isolate of Actinobacillus actinomycetemcomitans . This predicted protein had significant homology at the amino acid level to cation transport proteins . Front Biosci, 2001 Sep 01, 6, D1128 - 50 Molecular genetic analysis of virulence in Mannheimia (pasteurella) haemolytica; Highlander SK; Mannheimia haemolytica (previously known as Pasteurella haemolytica) is a weakly hemolytic, gram-negative coccobacillus that is an opportunistic pathogen of cattle, sheep and other ruminants . In stressed, immunocompromised animals, the organism causes a fibrinous, necrotic pneumonia, commonly called "shipping fever" . In the United States, economic losses due to shipping fever pneumonia surpass the combined cost of all other diseases of cattle . M . haemolytica, which is a member of the family Pasteurelleaceae, includes twelve serotypes (A1, A2, A5-A9, A12-14, A16 and A17) based on capsular antigen typing . Worldwide, serotypes A1 and A2 predominate, though all serotypes can cause disease . Serotype A1 causes pasteurellosis in cattle and has been the subject extensive study, while serotype A2 causes disease in sheep and is less-well characterized . Potential virulence factors of M . haemolytica have been identified and characterized by gene cloning and DNA sequence analysis . These factors include a ruminant-specific leukotoxin, an anti-phagocytic capsule, lipopolysaccharide, iron-regulated outer membrane proteins, lipoproteins, a sialoglycoprotease, a neuraminidase and two potential immunoglobulin proteases . Unlike the well-characterized leukotoxin, little is known about the expression of these other virulence factors . Attempts to dissect the mechanisms of M . haemolytica pathogenesis have been hindered by the lack of a robust genetic system for mutation of the organism, though new tools for genetic manipulation of M . haemolytica have been developed . Expression plasmids and operon fusion plasmids have been created and a series of antibiotic resistance cassettes useful for site-specific recombination have been constructed . It is anticipated that use of these tools for gene expression and mutagenesis, in combination with the soon to be released genomic sequence of a serotype A1 organism, will aid in understanding the molecular mechanisms of pathogenesis of M . haemolytica and will help to drive development of new vaccines to prevent shipping fever. J Endotoxin Res, 2001, 7(1), 49 - 52 A novel acute phase marker in cattle: lipopolysaccharide binding protein (LBP); Schroedl W et al.; The host response to infection, the "acute phase response" is a highly conserved series of physiological reactions including marked changes in concentrations of plasma proteins . These proteins have been shown to participate in the immune response to infections . Several recent studies have elevated the role of acute phase proteins (APPs) as predictive markers in infection . APPs such as serum amyloid A and haptoglobin but not C-reactive protein (CRP) have been identified as markers of inflammation in cattle . In humans, lipopolysaccharide (LPS) binding protein (LBP) has certain biological functions in host defence and participates in acute phase reactions . We measured plasma levels of LBP in a group of 20 calves experimentally infected with Gram-negative Mannheimia haemolytica (Pasteurella) in comparison to haptoglobin, the most widely studied APP in cattle . In infected calves, LBP levels rose significantly 6 h after infection, reaching a maximum at 24 h . Haptoglobin concentrations significantly rose after 12 h, and peak responses were measured 48 h after infection . Thus, LBP may prove to be a diagnostic marker in cattle infection and is faster than haptoglobin in detecting sepsis. Vet Microbiol, 2001 Oct 1, 82(4), 361 - 72 Random amplification of polymorphic DNA and phenotypic typing of Zimbabwean isolates of Pasteurella multocida; Dziva F et al.; Eighty-one isolates presumptively identified as Pasteurella multocida from a variety of diseases in animals in Zimbabwe were subjected to biochemical characterization, capsular typing and RAPD analysis . The majority of isolates (over 80%) were assigned into named taxa and were predominantly P . multocida subsp . multocida and P . multocida subsp . septica, whilst the remainder were unassigned . Serogroup A was predominant among the three capsular types (A, B and D) of P . multocida detected . Three main RAPD clusters and three subclusters were observed among the majority of isolates (93.8%), whilst the remainder was found to be weakly related . Nine different groups of strains with similar RAPD profiles (100% similarity) were also observed . The reference strain of capsular serogroup F clustered with the reference strain of P . multocida subsp . septica, whilst all other serogroups clustered with reference strains of subsp . multocida and gallicida . Notably, serogroups A and D were observed to be closely related to the reference strain of subsp . multocida . The relationship between biotype, capsular type, host origin and disease manifestation was not clear-cut . However, most pig isolates of subsp . multocida clustered together as did most cattle isolates of subsp . multocida . RAPD tended to separate subsp . multocida from septica. J Wildl Dis, 2001 Jul, 37(3), 413 - 26 Health protocol for translocation of free-ranging elk; Corn JL et al.; When considering an elk (Cervus elaphus) restoration program, wildlife managers must evaluate the positive and negative elements of translocation . We prepared this protocol to give an overview of health considerations associated with translocation of elk, with an emphasis on movement of free-ranging elk from western North America to the southeastern USA . We evaluated infectious agents and ectoparasites reported in elk from two perspectives . First, we made a qualitative estimate of the ability of the agent to be introduced and to become established . This was done using a selected set of epidemiologic factors . Second, if there was a good possibility that the organism could become established in the release area, the potential pathological consequences for elk and other wildlife, domestic animals, and humans were assessed via examination of the literature and consultation with other animal health specialists . The results of these evaluations were used to classify infectious agents and ectoparasites as low risk (n = 174), unknown risk (n = 10), and high risk (n = 9) . We classified Anaplasma marginale, Anaplasma ovis, Mycobacterium paratuberculosis, Pasteurella multocida serotype 3, Elaphostrongylus cervi, Dicrocoelium dendriticum, Fascioloides magna, Echinococcus granulosus, Dermacentor albipictus, and Otobius megnini as unknown risks . High risk infectious agents and ectoparasites were the agent of chronic wasting disease, Brucella abortus, Mycobacterium bovis, Dermacentor andersoni, Ixodes pacificus, and Psoroptes sp . Parelaphostrongylus tennis, Elaeophora schneideri, and a Babesia sp . are parasites endemic in the southeastern USA that may present a "reverse risk" and adversely affect elk if released in some parts of the region . We developed a five-component protocol to reduce the risk of introduction of high risk infectious agents and ectoparasites that included: (1) evaluation of the health status of source populations, (2) quarantines, (3) physical examination and diagnostic testing, (4) restrictions on translocation of animals from certain geographic areas or populations, and (5) prophylactic treatment. Infect Immun, 2001 Sep, 69(9), 5931 - 5 Escherichia coli cytotoxic necrotizing factor and Pasteurella multocida toxin induce focal adhesion kinase autophosphorylation and Src association; Thomas W et al.; Cytotoxic necrotizing factor 1 and Pasteurella multocida toxin induced dose- and time-dependent increases in focal adhesion kinase (FAK) Tyr397 phosphorylation in Swiss 3T3 cells . FAK autophosphorylation was sensitive to inhibitors of p160/ROCK and coincided with the formation of stable complexes between FAK and Src family members. Biologicals, 2001 Mar, 29(1), 7 - 10 Pasteurella multocida toxin type D serological assay as an alternative to the toxin neutralisation lethality test in mice; Finco-Kent DL et al.; The objective of this work was to develop an enzyme-linked immunosorbent assay (ELISA) for the detection of antibodies against Pasteurella multocida toxin type D, that correlated to a mouse lethality test . Currently, the mouse lethality test is one of several tests used world-wide to evaluate serological responses in animals immunised with vaccines containing toxoids . The mouse lethality test involves injecting mice with a mixture of toxin and test serum sample (from animals that have been vaccinated with a toxoid), and then determining antibody titre of the test serum from the number of mice that survive . Thus, the titre calculated is based on the neutralising activity of the test serum . The mouse lethality test requires large numbers of animals and causes severe distress to the animals . Organisations world-wide are working towards alternatives to animals in the development and control of biological products for human and veterinary use . Additionally, the mouse lethality test is labour-intensive, costly and lacks robustness and may be difficult to reproduce between different technicians . We have developed a double sandwich ELISA to measure anti- P . multocida toxoid type D antibodies in swine serum . Sera from swine immunised with vaccines containing type D toxoid showed good correlation to the mouse lethality assay (Spearman analysis=0.94 and Pearson analysis=0.84) . When compared to the mouse lethality test, titres obtained using the ELISA format had higher correlation with protective immunity (i.e., lower turbinate atrophy) following challenge with virulent P . multocida . The ELISA assay is more robust, reproducible and costs less than the mouse lethality assay; and it complements efforts to reduce the use of animals in testing . Can J Vet Res, 2001 Jul, 65(3), 181 - 7 Pharmacokinetics of orbifloxacin and its concentration in body fluids and in endometrial tissues of mares; Haines GR et al.; Pharmacokinetics and distribution of orbifloxacin into body fluids and endometrium was studied in 6 mares after intragastric (IG) administration at a single dose rate of 7.5 mg/kg body weight . Orbifloxacin concentrations were serially measured in serum, synovial fluid, peritoneal fluid, urine, cerebrospinal fluid, and endometrial tissues over 24 hours . Minimum inhibitory concentrations of orbifloxacin were determined for 120 equine pathogens over an 11-month period . The mean peak serum concentration (Cmax) was 2.41+/-0.30 microg/mL at 1.5 hours after administration and decreased to 0.17+/-0.01 microg/mL (Cmin) at 24 hours . The mean elimination half-life (t1/2) was 9.06+/-1.33 hours and area under the serum concentration vs time curve (AUC) was 20.54+/-1.70 mg h/L . Highest mean peritoneal fluid concentration was 2.15+/-0.49 microg/mL at 2 hours . Highest mean synovial fluid concentration was 1.17+/-0.28 microg/mL at 4 hours . Highest mean urine concentration was 536.67+/-244.79 microg/mL at 2 hours . Highest mean endometrial concentration was 0.72+/-0.23 microg/g at 1.5 hours . Mean CSF concentration was 0.46+/-0.55 microg/mL at 3 hours . The minimum inhibitory concentration of orbifloxacin required to inhibit 90% of isolates (MIC90) ranged from < or = 0.12 to > 8.0 microg/mL, with gram-negative organisms being more sensitive than gram-positive organisms . Orbifloxacin was uniformly absorbed in the 6 mares and was well distributed into body fluids and endometrial tissue . At a dosage of 7.5 mg/kg once a day, many gram-negative pathogens, such as Actinobacillus equuli, Escherichia coli, Pasteurella spp., and Salmonella spp . would be expected to be susceptible to orbifloxacin. Trop Anim Health Prod, 2001 Jul, 33(4), 275 - 83 Specific antibodies of Pasteurella multocida in newborn calves of vaccinated dams; el-Eragi AM et al.; Twenty-five newborn Holstein Friesian calves, from dams vaccinated against haemorrhagic septicaemia (HS), were tested repeatedly over the first 6 months of life to monitor the transferred antibody levels against HS . Enzyme-linked immunosorbent assays were used to measure the specific HS antibodies with antigens from Pasteurella multocida strains B:2 and E:2 . There was a significant curvilinear relationship between the monitored IgG response and the age of the calves . Peak serum IgG levels were obtained during the period from 8 to 16 weeks of age . Beyond this age, the concentration of IgG in the serum fell away. Poult Sci, 2001 Jun, 80(6), 689 - 94 Effect of selection for increased body weight in turkeys on lymphoid organ weights, phagocytosis, and antibody responses to fowl cholera and Newcastle disease-inactivated vaccines; Li Z et al.; The influence of selection was studied for increased 16-wk BW in turkeys on in vivo phagocytic activity, antibody responses to vaccines, and weight of the spleen and bursa of Fabricius . A line (F) of turkeys selected long term for increased 16-wk BW and its corresponding randombred control (RBC2) were compared . Phagocytic activity was evaluated by the carbon clearance assay . Antibody responses to inactivated Newcastle disease virus and Pasteurella multocida vaccines were examined by ELISA . Body weight and relative weights of spleen and bursa of Fabricius of the two lines were also compared . The F line had lower phagocytic activity than the RBC2 line (P < 0.05) . In addition, the F line had greater BW, relative weight of spleen, and ratio of spleen to bursa of Fabricius weight (P < 0.01) but had a lower relative weight of bursa of Fabricius at 9 wk of age . However, there were no line differences in the antibody responses to Newcastle disease virus or P . multocida vaccines at 1, 2, 3, 4, 5, or 12 wk after vaccination . Based on the present results, it is suggested that long-term selection for increased 16-wk BW might have resulted in changes in the immune system, as indicated by changes in the relative weights of the spleen and bursa of Fabricius and phagocytic activity . The decreased phagocytic activity in the F line may be partially responsible for increased susceptibility to specific diseases in this line. Vet Res, 2001 May-Aug, 32(3-4), 323 - 39 Antimicrobial resistance in pasteurella and mannheimia: epidemiology and genetic basis; Kehrenberg C et al.; Isolates of the genera Pasteurella and Mannheimia cause a wide variety of diseases of great economic importance in poultry, pigs, cattle and rabbits . Antimicrobial agents represent the most powerful tools to control such infections . However, increasing rates of antimicrobial resistance may dramatically reduce the efficacy of the antimicrobial agents used to control Pasteurella and Mannheimia infections . This review presents a short summary of the infections caused by Pasteurella and Mannheimia isolates in food-producing animals and the possibilities of preventing and controlling primary and secondary pasteurellosis . Particular reference is given to antimicrobial chemotherapy and the resistance properties of Pasterurella and Mannheimia isolates . The genetic basis of the most predominant resistance properties such as resistance to beta-lactam antibiotics, tetracyclines, aminoglycosides, sulfonamides, and chloramphenicol is discussed . This is depicted with reference to the role of plasmids and transposons in the spread of the resistance genes among Pasteurellaceae and members of other bacterial families and genera. J Clin Microbiol, 2001 Jul, 39(7), 2558 - 64 Pasteurella multocida subsp . multocida and P . multocida subsp . septica differentiation by PCR fingerprinting and alpha-glucosidase activity; Hunt Gerardo S et al.; Pasteurella multocida is composed of three subspecies that are often differentiated by fermentation of sorbitol and dulcitol . We studied 35 dulcitol-negative P . multocida isolates from infected dog and cat bite wounds, 16 of which yielded weak and/or conflicting fermentation reactions in Andrades sorbitol, thus making it difficult to distinguish between the two dulcitol-negative subspecies of P . multocida, i.e., P . multocida subsp . multocida and P . multocida subsp . septica . All isolates and two control strains were further analyzed using a PCR fingerprinting technique with a single primer (M13 core) and assessed for alpha-glucosidase (alpha-Glu) activity . Although the PCR fingerprint patterns and alpha-Glu activity did not correlate well with the sorbitol fermentation reactions, they did correlate well with each other . All strains identified as P . multocida subsp . septica were positive for alpha-Glu activity and exhibited the group I PCR fingerprint profile . All strains categorized as P . multocida subsp . multocida displayed either the group II or group III PCR fingerprint profile; 9 of 11 of these isolates were alpha-Glu negative . These data suggest that both PCR fingerprinting and alpha-Glu activity provide reliable means for differentiating P . multocida subsp . multocida from P . multocida subsp . septica, particularly in strains that produce weak and/or discrepant sorbitol fermentation reactions. Avian Dis, 2001 Apr-Jun, 45(2), 540 - 3 Infraorbital sinusitis associated with Pasteurella multocida in pen-raised ring-necked pheasants; Chin RP et al.; Pasteurella multocida, somatic serotype 6, was isolated from the infraorbital sinuses of 8-wk-old ring-necked pheasants with severe sinusitis . In addition, Escherichia coli, Pasteurella haemolytica-like bacteria, Mycoplasma gallinaceum, and Mycoplasma glycophilum were also isolated from some of the sinuses . Clinical signs appeared 3 days after placement on the grow-out ranch . The sinusitis consisted of severe unilateral or bilateral distention of the sinuses by mucoid to caseous exudate . Mortality and morbidity were low . Birds responded to treatment with tetracycline after proper medication procedures . The source of the infection was not determined, though possible sources include the brood ranch, wild animals, or wild waterfowl. Avian Dis, 2001 Apr-Jun, 45(2), 522 - 8 An outbreak of duck viral enteritis (duck plague) in domestic Muscovy ducks (Cairina moschata domesticus) in Illinois; Campagnolo ER et al.; Duck viral enteritis (DVE) was diagnosed in an outbreak of the disease in a resident population of Muscovy ducks (Cairina moschata domesticus) on a privately owned multispecies game bird production facility in Illinois, where it claimed 625 ducks . This disease condition had not been reported previously in domestic ducks in Illinois . Although other varieties and age groups of domestic waterfowl (i.e., black ducks, rhumen ducks, Pekin ducks, ducklings, and geese) were present on the game bird farm, the morbidity and mortality (100%) in this epornitic was solely limited to adult ducks of the Muscovy lineage . The clinical signs in the affected ducks were lethargy, diarrhea, dehydration, and death within 2-3 hr of onset of symptoms . Gross pathologic changes were nonspecific and included ecchymotic hemorrhage, effusion of fluid and blood within body cavities reflective of an acute systemic infectious disease . Light microscopic findings were necrosis of primarily digestive lining epithelium and variable lymphohistiocytic infiltration within mucosal and serosal connective tissues . Intranuclear inclusions resembling characteristic herpetic (i.e., Cowdry type A) inclusions were observed primarily in the digestive, respiratory, and reproductive tracts; liver; and spleen . Esophageal candidiasis, bacteriosis, and systemic Pasteurella anatipestifer infections, thought to be concurrent or opportunistic infections, were present in several ducks . DVE virus was demonstrated in infected Muscovy duck embryo fibroblast cells by direct DVE virus-specific fluorescent antibody staining. Berl Munch Tierarztl Wochenschr, 2001 May-Jun, 114(5-6), 169 - 72 Mastitis in lactating camels (Camelus dromedarius) in Afar Region, north-eastern Ethiopia; Bekele T et al.; Quarter milk samples (n = 543) from 152 traditionally managed lactating camels (Camelus dromedarius) in Afar Region, north-eastern Ethiopia were examined to determine the prevalence of camel mastitis and identify its bacterial causes . Out of 152 camels examined, 19 (12.5%) were diagnosed as clinical mastitis cases based on clinical signs and bacteriological examinations . Of the 257 California Mastitis Test (CMT) positive quarter milk samples 162 (63.0%) yielded pathogenic bacteria . A positive correlation was observed between CMT positive results and presence of major pathogens in camel milk samples . The main mastitis pathogens isolated were Staphylococcus aureus, coagulase-negative staphylococci, Streptococcus agalactiae, S . dysgalactiae, and other species of streptococci, Pasteurella haemolytica and E . coli . Results of the present study suggest that mastitis in Afar camels is prevalent, Gram-positive cocci are the major isolates from camel milk samples and the CMT can be used as a screening test for the detection of mastitis in camels. JAMA, 2001 Jun 6, 285(21), 2763 - 73 Tularemia as a biological weapon: medical and public health management; Dennis DT et al.; OBJECTIVE: The Working Group on Civilian Biodefense has developed consensus-based recommendations for measures to be taken by medical and public health professionals if tularemia is used as a biological weapon against a civilian population . PARTICIPANTS: The working group included 25 representatives from academic medical centers, civilian and military governmental agencies, and other public health and emergency management institutions and agencies . EVIDENCE: MEDLINE databases were searched from January 1966 to October 2000, using the Medical Subject Headings Francisella tularensis, Pasteurella tularensis, biological weapon, biological terrorism, bioterrorism, biological warfare, and biowarfare . Review of these references led to identification of relevant materials published prior to 1966 . In addition, participants identified other references and sources . CONSENSUS PROCESS: Three formal drafts of the statement that synthesized information obtained in the formal evidence-gathering process were reviewed by members of the working group . Consensus was achieved on the final draft . CONCLUSIONS: A weapon using airborne tularemia would likely result 3 to 5 days later in an outbreak of acute, undifferentiated febrile illness with incipient pneumonia, pleuritis, and hilar lymphadenopathy . Specific epidemiological, clinical, and microbiological findings should lead to early suspicion of intentional tularemia in an alert health system; laboratory confirmation of agent could be delayed . Without treatment, the clinical course could progress to respiratory failure, shock, and death . Prompt treatment with streptomycin, gentamicin, doxycycline, or ciprofloxacin is recommended . Prophylactic use of doxycycline or ciprofloxacin may be useful in the early postexposure period. Acta Vet Hung, 2000, 48(4), 387 - 95 Clinical study of the disease of calves associated with Mycoplasma bovis infection; Stipkovits L et al.; Clinical, bacteriological and serological examination of 35 calves from the age of 5 to 26 days was performed in a Holstein-Friesian dairy herd endemically infected with Mycoplasma bovis . M . bovis was isolated from 48.6% of nasal swabs taken from the calves at the age of 5 days, and from 91.4% of the same calves at the age of 26 days, indicating the gradual spread of infection . The isolation rate of Pasteurella multocida did not change much, and varied from 28.6 to 25.7% . No P . haemolytica could be detected . In addition to M . bovis and P . multocida, the herd was also infected with different viruses (including bovine viral diarrhoea virus, infectious bovine rhinotracheitis virus, bovine adenoviruses, parainfluenza-3 virus, and bovine respiratory syncytial virus) as a large proportion of the sera of newborn calves contained colostral antibodies against these viruses . In most of the newborn calves severe clinical signs (fever, depression, inappetence, hyperventilation, dyspnoea, nasal discharge and coughing) due to M . bovis infection developed . The clinical signs appeared already on the fifth day of life, and their incidence was the highest at the age of 10 to 15 days . Three calves (8.6%) died as a result of severe serofibrinous pneumonia . The surviving calves showed very poor weight gain (ranging from 1.5 to 3.5 kg) during the first two weeks of life. Microb Pathog, 2001 Jun, 30(6), 347 - 57 Lipopolysaccharide enhances cytolysis and inflammatory cytokine induction in bovine alveolar macrophages exposed to Pasteurella (Mannheimia) haemolytica leukotoxin; Lafleur RL et al.; Pasteurella (Mannheimia) haemolytica leukotoxin (Lkt) and lipopolysaccharide (LPS) are the primary virulence factors contributing to the pathogenesis of lung injury in bovine pneumonic pasteurellosis . Previous studies have characterized in vitro responses of bovine alveolar macrophages (AMs) to Lkt and LPS . Activation of AMs with Lkt or LPS causes induction of proinflammatory cytokines, and Lkt causes cytolysis of AMs at higher concentrations . Since AMs are exposed to both of these bacterial virulence factors during disease, previous studies may have underestimated the possibility of functional interactions between Lkt and LPS . The purpose of this study was to characterize the effect of simultaneous exposure to both Lkt and LPS on AM cytolysis and proinflammatory cytokine expression . Using cellular leakage of lactate dehydrogenase as an indirect measure of cytolysis, we studied AM responses to Lkt alone, LPS alone and Lkt+LPS . We found that 80-200 pg/ml LPS, which does not itself cause cytolysis, synergistically enhanced the cytolysis induced by 2-5 Lkt units (LU)/ml Lkt . Northern blot analysis demonstrated that synergism between Lkt and LPS resulted in increased levels of IL-8 mRNA, and that the kinetic patterns of TNF-alpha and IL-8 mRNA expression induced by Lkt+LPS differed from those induced by each agent separately . Finally, the WEHI 164 (clone 13) bioassay was used to show that Lkt/LPS synergism resulted in enhanced secretion of biologically active TNF-alpha . These results provide direct evidence of synergism between Lkt and LPS in AM cytolysis and inflammatory cytokine expression . Additional studies to characterize the molecular basis of this phenomenon are indicated . Microb Pathog, 2001 Jun, 30(6), 325 - 35 Mast cell density and substance P-like immunoreactivity during the initiation and progression of lung lesions in ovine Mannheimia (Pasteurella) haemolytica pneumonia; Ramirez-Romero R et al.; To determine the density of mast cells (MCs) and the extent of substance P (SP) immunoreactivity during initiation and progression of pneumonic pasteurellosis (PP), 18 lambs were inoculated intrabronchially with Mannheimia (Pasteurella) haemolytica or saline, and lung tissue was collected at 1, 15 and 45 days post-inoculation (n=3, each group) . Additionally, the left (non-inoculated) contralateral lungs in bacteria-inoculated animals were collected as controls . At 1 day after bacterial inoculation the lungs had typical M . haemolytica lesions . These pneumonic lesions had fewer numbers of MCs and reduced histamine content . Macrophages infiltrating some of the inflamed areas were strongly immunoreactive for SP . At 15 days, MCs remained scarce at sites where lung damage persisted, i.e . pyogranulomatous foci, but were increased in number in areas of interstitial damage . Pulmonary ganglion neurons were strongly immunoreactive for SP . By 45 days the fibrosing changes became more defined as pleural fibrosis, fibrosing alveolitis, alveolar epithelial hyperplasia and bronchiolitis obliterans . These lungs had increased numbers of MCs, but histamine content was not different from saline- and non-inoculated left lungs . Substance P immunoreactivity occurred only in nerves and was scarce and mild . This work demonstrates that MC density decreases initially with PP, but increases with progression of PP . SP fibres tend to be decreased during the initiation and at 45 days of PP, but other cells, such as macrophages and neuronal ganglion cells, produce substance P during progression of PP and thereby constitute an additional source of substance P . Int J Antimicrob Agents, 2001 Jun, 17(6), 505 - 10 Suppression of Mannheimia (Pasteurella) haemolytica serovar 1 infection in lambs by intrapulmonary administration of ovine antimicrobial anionic peptide; Kalfa VC et al.; In this study, the efficacy of ovine antimicrobial anionic peptide (AP) was assessed in a lamb model of acute pneumonia . A single intratracheal dose of the peptide, H-DDDDDDD-OH (0.5 mg) reduced pulmonary inflammation and the concentration of Mannheimia (Pasteurella) haemolytica in infected lung tissue . Administration of H-DDDDDDD-OH after infection was more effective in reducing the consolidation and lesion scores at the deposition site than its administration prior to infection . Hence, the in vivo effectiveness of AP suggests that it may have applications in the treatment of pulmonary infections . Further studies are needed to confirm these findings and also to determine the optimal doses and intervals of H-DDDDDDD-OH therapy. Int J Antimicrob Agents, 2001 Jun, 17(6), 471 - 6 Postantibiotic effects and postantibiotic sub-MIC effects of erythromycin, roxithromycin, tilmicosin, and tylosin on Pasteurella multocida; Lim J et al.; When intermittent dosing is used during treatment, the concentrations of antibiotics fluctuate and subinhibitory concentrations may occur between doses . Postantibiotic effects (PAEs) and postantibiotic subinhibitory effects (PA SMEs) on bacteria may provide additional, valuable information for the rational use of a drug in clinical practice . In this study tilmicosin was the most active antibiotic tested against P . multocida type D with MICs ranging from 4-16 mg/l . Roxithromycin and tilmicosin induced a statistically significantly longer PAE than did tylosin against P . multocida types A and D (P < 0.05) . The duration of PAEs and PA SMEs were proportional to the concentrations of drugs used for exposure . The PA SMEs were substantially longer than PAEs on P . multocida . Tilmicosin had a longer PA SME compared with erythromycin, roxithromycin and tylosin for P . multocida . The computerized incubator used in the present study provided an efficient and convenient determination of PAE and PA SME, allowing frequent measurements of the bacterial growth. Vet Microbiol, 2001 Aug 20, 81(4), 305 - 14 Survival of Mannheimia (Pasteurella) haemolytica in tracheobronchial washings of sheep and cattle; Rowe HA et al.; The growth, morphology and long-term survival of a representative isolate of Mannheimia haemolytica serotypes A1 and A2 were monitored in ovine and bovine tracheobronchial washings . Both strains survived for at least 244 days in ovine tracheobronchial washings and 156 days in bovine tracheobronchial washings . The addition of fresh washings at these times prompted an increase for serotype A2 but no change in viability for serotype A1 in ovine tracheobronchial washings and an increase for both serotypes in bovine tracheobronchial washings . When growth and survival was compared using tracheobronchial washings from ruminant and non-ruminant species there was a trend towards longer survival in ruminant fluids.Long-term survival was associated with temporary or permanent change from normal size colonies to 'micro-colonies' on sheep blood agar . Subculture allowed reversion to normal colony morphology . Analysis showed these micro-colonies to consist of chains of elongated bacteria . M . haemolytica serotype A2 was more robust in its ability to withstand nutrient deprivation for long periods of time . These survival mechanisms may have important implications for pathogenesis. Vet Pathol, 2001 May, 38(3), 297 - 310 Pulmonary expression of tumor necrosis factor alpha, interleukin-1 beta, and interleukin-8 in the acute phase of bovine pneumonic pasteurellosis; Malazdrewich C et al.; Inflammatory cytokines are suspected to contribute to the pathogenesis of bovine pneumonic pasteurellosis (BPP) through neutrophil recruitment, leukocyte activation, and the induction of a broad array of soluble inflammatory mediators . An in vivo experimental model of BPP was used to characterize the pulmonary expression kinetics of tumor necrosis factor alpha (TNFalpha), interleukin-1 beta (IL-1beta), and interleukin-8 (IL-8) genes and proteins during the acute phase of disease development . Cytokine expression in bronchoalveolar lavage (BAL) fluid, BAL cells, and pneumonic lung parenchyma was quantitated by northern blot analysis, enzyme-linked immunosorbent assay (ELISA), and in situ hybridization at 2, 4, 8, 16, and 24 hours after endobronchial inoculation of Pasteurella (Mannheimia) haemolytica . Expression of TNFalpha, IL-1beta, and IL-8 was significantly increased in the airways and lung lesions of infected calves as compared with mock-infected controls . Although kinetic patterns varied, peak levels of cytokine mRNA occured within 8 hours postinfection (PI), and peak cytokine concentrations occurred within 16 hours PI . In all samples, IL-8 was expressed to the greatest extent and TNFalpha was least expressed . Expression of TNFalpha was restricted to alveolar macrophages . Alveolar and interstitial macrophages produced IL-1beta and IL-8 in the first 4 hours; bronchial and bronchiolar epithelial cells were also significant sources of IL-8 during this period . By 8 hours PI, neutrophils were the dominant source of both IL-1beta and IL-8 . These findings demonstrate a spatial and temporal association between pulmonary expression of inflammatory cytokines and acute lung pathology, supporting the hypothesis that cytokines contribute to inflammatory lung injury in BPP. Equine Vet J, 2001 May, 33(3), 256 - 64 A case-control study of respiratory disease in Thoroughbred racehorses in Sydney, Australia; Christley RM et al.; In order to investigate the role of infectious agents in the aetiology of lower respiratory tract disease in Thoroughbred racehorses, a matched case-control study was conducted . Cases were identified by the presence of coughing, and were compared to a control population matched on time of sample collection and location within the same training establishment . Tracheal wash samples were collected from 100 cases and 148 controls . Case horses were more likely than controls to have endoscopic and cytological evidence of airway inflammation . There was no significant association between serological evidence of infection by commonly implicated respiratory viruses and coughing . Similarly, mycoplasma were rarely isolated and were not associated with disease . In contrast, there was a strong association between isolation of greater than a total of 10(3) colony-forming units/ml of tracheal wash and coughing . Individual bacterial species associated with disease included Streptococcus zooepidemicus, Streptococcus pneumoniae, Streptococcus suis, Streptococcus sanguis, Pasteurella spp and Bordetella bronchiseptica . This study provides evidence of the role of bacterial infection in the aetiology of lower respiratory tract inflammation in racehorses . However, in 58% of cases, few or no bacteria were isolated . Hence, at the time of identification of disease, there was no evidence of viral, bacterial or mycoplasmal infection in the majority of coughing horses . The aetiology of the signs observed in these horses requires further investigation. Infect Immun, 2001 Jun, 69(6), 4109 - 15 Pasteurella multocida gene expression in response to iron limitation; Paustian ML et al.; Pasteurella multocida is the causative agent of a wide range of diseases in avian and mammalian hosts . Gene expression in response to low iron conditions was analyzed in P . multocida using whole-genome microarrays . The analysis shows that the expression of genes involved in energy metabolism and electron transport generally decreased 2.1- to 6-fold while that of genes used for iron binding and transport increased 2.1- to 7.7-fold in P . multocida during the first 2 h of growth under iron-limiting conditions compared with controls . Notably, 27% of the genes with significantly altered expression had no known function, illustrating the limitations of using publicly available databases to identify genes involved in microbial metabolism and pathogenesis . Taken together, the results of our investigations demonstrate the utility of whole-genome microarray analyses for the identification of genes with altered expression profiles during varying growth conditions and provide a framework for the detailed analysis of the molecular mechanisms of iron acquisition and metabolism in P . multocida and other gram-negative bacteria. Infect Immun, 2001 Jun, 69(6), 3628 - 34 Biological activity of a C-terminal fragment of Pasteurella multocida toxin; Busch C et al.; The protein toxin of Pasteurella multocida PMT is a potent mitogen and activator of phospholipase Cbeta . In this study different toxin fragments were investigated . A C-terminal fragment encompassing amino acids 581 through 1285 (PMT581C) was constructed, which was inactive toward intact embryonic bovine lung (EBL) cells after addition to culture medium but caused reorganization of the actin cytoskeleton and rounding up of cells when introduced into the cells by electroporation . As the holotoxin, the toxin fragment PMT581C induced an increase in total inositol phosphate levels after introduction into the cell by electroporation . A C-terminal fragment shorter than PMT581C as well as N-terminal fragments were inactive . Exchange of cysteine-1165 for serine in the holotoxin resulted in a complete loss of the ability to increase inositol phosphate levels . Correspondingly, the mutated toxin fragment PMT581C.C1165S was inactive after cell introduction by electroporation, suggesting an essential role of Cys-1165 in the biological activity of the toxin. Eur J Clin Microbiol Infect Dis, 2001 Mar, 20(3), 210 - 3 Molecular identification of TEM-1 beta-lactamase in a Pasteurella multocida isolate of human origin; Naas T et al.; Two clinical strains of Pasteurella multocida were isolated from an HIV-infected patient who developed arthritis . Strain FB-1, which was isolated from a dog-bite wound, was resistant to narrow-spectrum penicillins . The second strain, FB-2, which was present in blood cultures as well as the dog-bite wound, was susceptible to all beta-lactam agents . Arbitrarily primedpolymerase c