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Effects of Pasteurella multocida toxin on porcine bone marrow cell differentiation into osteoclasts and osteoblasts.
The effect of Pasteurella multocida toxin (PMT) on porcine osteoclast and osteoblast differentiation was studied using in vitro cell culture systems. When grown in the presence of Vitamin D3, isolated porcine bone marrow cells formed multinucleated cells with features characteristic of osteoclasts. Exposure of bone marrow cells to Vitamin D3 and PMT during growth resulted in formation of increased numbers and earlier appearance of osteoclasts compared to controls. Ultrafiltered medium form PMT-treated cells likewise increased osteoclast numbers, suggesting that a soluble mediator may be involved in the action of PMT. When cell cultures were treated with fluorescein-labeled PMT, fluorescence was found within the cytoplasm of small, round cells that did not resemble either osteoclasts or osteoclastic precursor cells. Cultures of porcine bone marrow cells exposed to dexamethasone, ascorbic acid, and beta-glycerophosphate developed into osteoblastic cells that formed multilayered, mineralized nodules. Exposure of osteoblastic cultures to low concentration of PMT resulted in retarded cell growth, formation of decreased numbers of nodules and minimal to no mineralization in the nodules; higher concentration of PMT resulted in increased cellular debris and poor growth of cells, with no nodule formation. These findings suggest that PMT may induce turbinate atrophy in pigs by increasing osteoclast numbers and inhibiting osteoblastic bone formation. The effect of PMT on osteoclastic differentiation and growth may not be due to a direct effect on preosteoclastic cells, but rather due to alterations in the soluble mediator secretion by marrow stromal cells.
Syphilitic aortic aneurysm and squamous cell carcinoma of the penis: a case report.
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Evaluation of Pasteurella multocida isolated from rabbits by capsular typing, somatic serotyping, and restriction endonuclease analysis.
The goal of this study was to characterize Pasteurella multocida isolated from rabbits. Five hundred and fifty-three apparently healthy rabbits were sampled for this study. Nasal swabs were collected from each rabbit for P. multocida isolation and identification. Isolates were further characterized by capsular and somatic antigens and genomic DNA fingerprinting. Thirty-nine P. multocida isolates were recovered from 553 rabbits (7%). Capsular typing was done by depolymerization of P. multocida capsule by Staphylococcus aureus hyaluronidase and by disc diffusion with mucopolysaccharidase enzymes (heparinase III, chondroitinase AC, and hyaluronidase). Thirty-one (79%) of the isolates were capsular type A, and 8 isolates (21%) had untypable (UT) capsules. The gel-diffusion precipitin test was used to determine the somatic type of P. multocida isolates. Nineteen isolates were somatic serotype 3 (49%), 12 were serotype 1 (31%), 1 was serotype 2, 2 were serotype 5, 2 were serotype 12 with a weak reaction to antiserum raised against serotype 7 (5%), and 1 was serotype 4. Two of the isolates (5%) were UT. Restriction endonuclease analysis of the DNA of the isolates revealed 7 distinct profiles by digestion with HindIII, and 12 profiles were obtained with HpaII, whereas digestion with EcoRI did not differentiate between any of the P. multocida DNA isolates studied. The DNA restriction endonuclease enzyme HpaII was found more useful for differentiating between DNA fingerprints of P. multocida rabbit isolates. However, no correlation between capsular type, somatic serotypes, and DNA fingerprints was seen in this study.
Molecular epidemiology of two fowl cholera outbreaks on a free-range chicken layer farm.
Two outbreaks of fowl cholera on a multiage free-range egg farm were investigated. The outbreaks occurred in 1994 and 2002. A total of 22 strains of Pasteurella multocida were available for study, 11 from the 1994 outbreak and 11 from the 2002 outbreak. Lesions typical of acute fowl cholera were seen in the 1994 outbreak, whereas both acute and chronic fowl cholera occurred in the 2002 outbreak. The isolates were examined in an extended phenotypic typing methodology, by a P. multocida-specific polymerase chain reaction (PCR), by the Heddleston somatic serotyping scheme, and by restriction endonuclease analysis (REA) typing using the enzyme HpaII. All 22 strains had the same phenotypic properties, all were confirmed as P. multocida by PCR, all were Heddleston serovar 4, and all had the same REA pattern. The results indicate that these 2 outbreaks were caused by the same clone of P. multocida--despite the 8-year time period between the outbreaks.
Distribution of indole-producing urease-negative pasteurellas in animals.
Three hundred fifty-six animal isolates of indole-positive urease-negative cultures of Pasteurella, which would formerly have been classified as P. multocida, were examined with respect to their relationship to the recently described P. multocida subspecies (ssp.) multocida, septica, and gallicida and P. canis, P. stomatis/Taxon 16, and Pasteurella sp. B. Two hundred sixty-three (73.9%) of the cultures could be identified with one of these taxa, and 93 isolates (26.1%), representing 17 different biotypes, were unassignable. Pasteurella multocida ssp. multocida was the predominant taxon throughout and in most of the 25 animal species from which isolations were made. In dogs, P. canis was the most frequent. Different degrees of host predilection were observed also in P. multocida ssp. septica for cats, P. canis for sheep, and 2 of the unassignable biotypes for cattle and dogs, respectively. Overall, the respiratory tract was the most frequent source of isolates, but a propensity of P. multocida ssp. septica for localization in the central nervous system of cats was noted.
Use of ELISA to detect toxigenic Pasteurella multocida in atrophic rhinitis in swine.
The use of an enzyme-linked immunosorbent assay (ELISA) as a means of detecting dermonecrotoxin-producing strains of Pasteurella multocida was investigated. The assay was evaluated as a means to identify toxigenic P. multocida isolates recovered from nasal secretions of swine with atrophic rhinitis. The sensitivity and specificity of the ELISA for detecting dermonecrotoxin-producing P. multocida strains were compared to those of mouse-inoculation and cytotoxicity assays. The ELISA was highly sensitive and more specific than animal inoculation or tissue culture assay and is thus a more effective method for screening swine herds for the presence of toxigenic strains of P. multocida. The ELISA is a rapid, effective, economical way to identify toxigenic P. multocida isolates.
Minimum inhibitory concentration breakpoints and disk diffusion inhibitory zone interpretive criteria for tilmicosin susceptibility testing against Pasteurella spp. associated with bovine respiratory disease.
Tilmicosin is a novel macrolide antibiotic developed for exclusive use in veterinary medicine. The first tilmicosin-containing product was approved to treat bovine respiratory disease associated with pasteurellae. The development of antimicrobial susceptibility testing guidelines for tilmicosin was predicated on the relationship of clinical efficacy studies that demonstrated a favorable therapeutic outcome, on pharmaco-kinetic data, and on in vitro test data, as recommended by the National Committee for Clinical Laboratory Standards (NCCLS). The NCCLS-approved breakpoints for the MIC dilution testing are resistant > or = 32 micrograms/ml, intermediate 16 micrograms/ml, and susceptible < or = 8 micrograms/ml. The zone of inhibition interpretive criteria for disk diffusion testing with a 15-micrograms disk are resistant < or = 10 mm, intermediate 11-13 mm, and susceptible > or = 14 mm.
Evaluation of techniques for the detection of toxigenic Pasteurella multocida strains from pigs.
Recently acquired field isolates and archived isolates from our collection of Pasteurella multocida were analyzed for production of dermonecrotic toxin. Detection of the toxin was carried out using a fetal lung feline (FLF) cell line and a commercial enzyme-linked immunosorbent assay (ELISA) kit. The dermonecrotic toxin gene (ToxA) was also detected using a polymerase chain reaction (PCR) technique. Results from the 3 methods were compared. Field isolates (group 1) came from a commercial herd that had clinical signs of atrophic rhinitis. Fifty-six (17.9%) strains were isolated from 312 nasal swabs. Thirty-five of these strains belonged to serotype A and the rest (21/56), although probably serotype D, were not characterized further. All of these strains were toxin negative based on both the ELISA and FLF cell culture results. Five isolates gave faint bands in the PCR reaction, and the rest (51/56) were PCR negative. PCR and ELISA were also performed from the initial swab cultures (mixed cultures); 7 samples gave faint PCR bands, but ELISA results were all negative. Archived strains (group 2) had been isolated from clinical cases of atrophic rhinitis and from cases of pulmonary pasteurellosis. A total of 76 strains were analyzed; 46 were serotype A, and the rest (30) were serotype D. ELISA and FLF cell culture tests were negative for all serotype A strains; however, 3 strains showed faint bands in the PCR reaction. Fourteen serotype D strains showed positive results in both the ELISA and the FLF cell culture tests. PCR from these samples also gave positive results showing a strong band in the gel. However, 4 strains that were ELISA and FLF cell culture negative showed a faint band in the PCR reaction. The 3 methods gave similar results in the detection of the P. multocida dermonecrotic toxin. However, complete agreement among the tests was achieved only when strong PCR bands were considered positive. This is the first report that demonstrates the use of FLF cell line for the detection of toxigenic P. multocida.
Pasteurella multocida in peritoneal dialysis: a rare cause of peritonitis associated with exposure to domestic cats.
Pasteurella multocida is a rare cause of peritonitis in peritoneal dialysis patients with only 10 cases reported in the literature so far. All cases were observed in patients with close contact with cats, usually with a direct puncture of the dialysis tubing. We report a case of Pasteurella multocida peritonitis in a patient maintained under continuous cycling peritoneal dialysis (CCPD), who had frequent and close contact with cats. Patients should be made aware of this potential complication and advised to keep domestic animals away from the location of their peritoneal exchanges.
Pancoast's syndrome due to chronic pneumonia by Pasteurella multocida.
Infectious causes of Pancoast's syndrome are extremely rare. We describe the case of a patient with Pancoast's syndrome due to chronic pneumonia resulting from Pasteurella multocida. The patient was not immunosuppressed and had had no contact with animals. The diagnosis was made by transthoracic needle aspiration and institution of therapy with cefuroxime-axetil resulted in resolution of his symptoms.
SEPPIC vaccine adjuvants for poultry.
Two inactivated antigens (Newcastle and Pasteurella Multocida) were formulated with different adjuvants and tested in two separate experiments in poultry. Oil formulations constituting water in oil (W/O) or water in oil in water (W/O/W) emulsions were assessed for antibody response, protection, local reactions, and vaccine physicochemical parameters. Robust, efficacious, and safe formulations were obtained with W/O formulations whereas W/O/W was especially safe with maintained efficacy. Results show that it is possible to improve traditional Tween Span formulations for safety and efficacy parameters by using Montanide ISA 70 for W/O formulations and Montanide ISA 206 for W/O/W when safety is the priority.
Immunization of rabbits against Pasteurella multocida using a commercial swine vaccine.
Elaboration of heat-labile toxin (PMT) is an important virulence factor in some isolates of Pasteurella multocida from rabbits. Previously, we reported that immunization with inactivated PMT (IPMT) stimulated protective immunity to challenges from PMT. To test the hypothesis that immunization with a commercial swine vaccine containing IPMT stimulates similar protective immunity, groups of five rabbits were inoculated twice intramuscularly (i.m.), 10 days apart, with 0.5 ml of sterile saline or a commercial swine P. multocida bacterin-toxoid (BT). In addition, a group was inoculated intranasally with 5 microg of IPMT. Serum and nasal lavage samples were taken on days 0, 7, 14 and 21 after initial immunization and assayed by ELISA for anti-PMT antibody. Serum IgG and nasal lavage IgA were detectable by day 14 in BT and IPMT-immunized rabbits, but not in the saline controls. Groups of similarly inoculated rabbits were then challenged intranasally with 28 microg of PMT 21 days after initial immunization, and necropsied 7 days later, along with control challenged and non-challenged rabbits. Histological lesion severity was graded on a numerical scale. Non-immunized and saline, challenged controls developed more severe pneumonia, pleuritis, nasal turbinate atrophy and testicular atrophy than IPMT and BT-immunized rabbits. The results confirm the hypothesis that immunization with a commercial swine P. multocida BT confers protective immunity in rabbits against challenges from PMT.
Pathogenicity of Pasteurella multocida A:3 in Flemish giant and New Zealand white rabbits.
Pasteurella multocida A:3 was isolated during an outbreak of pasteurellosis in Flemish Giant (FG) rabbits. Since New Zealand White (NZW) rabbits housed in the same room were not as severely affected as FG rabbits, experimental inoculation was undertaken to determine if FG rabbits were more susceptible than NZW rabbits to pasteurellosis induced by this isolate. Rabbits of each breed were inoculated with P. multocida A:3 and observed for 3 weeks. Four of 5 FG rabbits developed severe clinical disease on days 6, 9, 12 and 14 after inoculation; whereas, the one affected NZW rabbit became ill 14 days after inoculation. All rabbits with clinical disease developed fibrinosuppurative pleuritis, pyothorax and pneumonia which was more severe in FG than NZW rabbits. At necropsy, P. multocida A:3 was isolated from multiple sites of the diseased rabbits. No significant difference (P = 0.099) in the prevalence of lesions between the two breeds was found; however, the score of pneumonia and pleuritis was 3 times greater in FG rabbits than NZW rabbits.
The first outbreak of fowl cholera in Muscovy ducks (Cairina moschata) in Japan.
The first outbreak of fowl cholera occurred in a flock of Muscovy ducks (Cairina moschata) in Okinawa Prefecture of Japan in November 1990. Fifty (25%) of 200 birds in a farm died of an acute disease. Remaining birds recovered after treatment with oxytetracycline. Pasteurella multocida subsp. multocida was isolated in pure culture from all tissues tested from two dead birds. Serovars of the isolates were identified as Carter's capsular type A. Heddleston's type 3.4.12, and Namioka's type 5:A which have not been demonstrated in Japan. Pathologically, multiple necrosis and bacterial aggregates were prominent in several organs, particularly in the liver. The isolate killed chickens when inoculated intravenously at a concentration of 10(8) colony forming units.
Toxigenic type A Pasteurella multocida as a causative agent of nasal turbinate atrophy in swine.
Although no clinical signs of atrophic rhinitis (AR) were recognized in 2- and 5-week-old pigs, approximately 60% of 2- to 6-month-old pigs showed clinical signs of AR in an affected pig farm. None of the pigs had normal turbinate at slaughter. Bordetella bronchiseptica was not isolated from any of the pigs before onset and incipient stage of the outbreak (2-week to 2-month-old). Pasteurella multocida of capsular type D was not isolated from any of those pigs. However, toxigenic P. multocida of capsular type A was isolated from a number of the pigs immediately before onset and incipient stage of the outbreak. Thirty-six-day-old primary specific-pathogen-free pigs were inoculated intranasally with a toxigenic type A P. multocida isolated from a 5-week-old pig. Severe nasal turbinate atrophy was observed in those pigs which were necropsied at 3 weeks post-inoculation. This is the first report on outbreak of severe nasal turbinate atrophy induced by toxigenic type A P. multocida in Japan.
Protectivity of an immunoaffinity-purified 39 kDa capsular protein of avian Pasteurella multocida in mice.
A 39 kDa protein of avian Pasteurella multocida strain P-1059 (serovar A:3) was purified from a crude capsular extract by immunoaffinity chromatography by using a ligand of purified mouse monoclonal antibody to the 39 kDa capsular protein of the strain. Protective activity of the purified 39 kDa protein antigen was determined by inoculation of ddY mice twice with 25 or 125 mug of the protein and challenge-exposure with 10 or 50 LD(50) of strains P-1059 or X-73 (serovar A:1). The results showed that the antigen gave high protection (60 to 100%). These results indicated that the 39 kDa protein of avian P. multocida is a cross-protective antigen over serovars A:1 and A:3.
The effects of early chest tube placement on empyema resolution.
STUDY OBJECTIVES: The objective of this study was to determine the impact of the timing of chest tube insertion on outcome for the treatment of empyema, using a new animal model of empyema. DESIGN: A prospective, controlled randomized, blinded design was used. SETTING: The study was conducted in an animal research laboratory. PATIENTS OR PARTICIPANTS: Sixty-six 2- to 3-kg rabbits were used in this study. INTERVENTIONS: After induction of empyema, the rabbits were divided into four groups. Fourteen rabbits had chest tubes placed at 24 h after empyema induction. Seventeen rabbits had chest tubes placed at 48 h and 14 rabbits had chest tubes placed at 72 h after empyema induction. Twenty-one rabbits served as control rabbits and had no chest tubes placed. MEASUREMENTS AND RESULTS: Ten days after induction of empyema, the rabbits were killed. The pleural spaces of each rabbit were examined and a gross score, pleural peel score, and a microscopic score were calculated for each rabbit. The median gross score, mean pleural peel score, and median microscopic scores were significantly higher in the rabbits that underwent late chest tube placement (72 h) relative to those that underwent early chest tube placement (24 or 48 h). CONCLUSIONS: This study supports previous expert opinion statements and conclusions from retrospective analyses that early chest tube placement (relative to delayed chest tube placement) is beneficial for the treatment of empyema.