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Experimentally induced infections bovine keratoconjunctivitis: effectiveness of a pilus vaccine against exposure to homologous strains of Moraxella bovis.

A study was conducted to determine whether vaccination with sonicated pili of Moraxella bovis would protect cattle from subsequent infection and disease when experimentally challenged by exposure to homologous cultures of M bovis. Some calves were intramuscularly inoculated 2 times with pili of M bovis suspended in water, and others were subcutaneously inoculated 2 times with pili of M bovis suspended in oil; 21 days were allowed between inculations. Controls were nonvaccinated calves. Fourteen days after the last inculation, all calves were exposed to virulent homologous cultures of M bovis. The results indicated that vaccination with sonicated pili of M bovis may induce protective immunity against homologous strain challenge exposure. Vaccines in oil that were injected subcutaneously protected to a greater extent than did vaccines in water that were injected intramuscularly. The development of inflammatory nodules at the site of inoculation was associated with resistance to infection and disease. Only 1 of the vaccinated calves that resisted disease lacked precipitating antibodies against sonicated pili at the time of the challenge exposure. This calf had antibodies 2 weeks later.

Animals↗

Induced infectious bovine keratoconjunctivitis: vaccination with whole cell bacterins of Moraxella bovis mixed with Freund's incomplete adjuvant.

Our studies on vaccination against infectious bovine keratoconjunctivitis were extended to include the use of adjuvant bacterins. Seventeen calves were given 2 doses of killed Moraxella bovis mixed with Freund's incomplete adjuvant; 15 calves were given 2 doses of whole cell bacterin without adjuvant. Four weeks later, all vaccinated and non-vaccinated (control) calves were challenge exposed by the conjunctival administration of virulent M bovis. The incorporation of Freund's incomplete adjuvant did not enhance the immunogenicity of M bovis bacterin for protection of calves against infectious bovine keratoconjunctivitis. Results indicated that the use of Freund's incomplete adjuvant does not compensate for a reduction in the number of doses which are required for protective vaccination of cattle.

Animals↗

[Recent trends in incidence of respiratory tract pathogens and antimicrobial susceptibilities of Haemophilus influenzae, Streptococcus pneumoniae and Moraxella catarrhalis isolated in 1994 and 1995].

The incidence of pathogenic bacteria in respiratory tract infections in 1994 and 1995 was investigated using quantitative cultures of sputa from patients with the infections in our department. Haemophilus influenzae, Streptococcus pneumoniae and Moraxella catarrhalis were isolated at high rates (70.5% in 1994 and 73.8% in 1995) from the specimens of out-patients, and the incident rates were similar to the past data. The antimicrobial susceptibilities of these three pathogens were examined with the agar dilution method. The incidence of penicillin (Pc) resistant S. pneumoniae against which MIC of Pc-G was higher than 0.125 microgram/ml was markedly increased from 24% in 1994 to 34.9% in 1995. Most of the Pc resistant isolates were also resistant to other antibiotics including erythromycin, minocycline and tosufloxacin. Serotype of strains against which MIC of Pc-G was higher than 1.0 microgram/ml was 19. The ratios of beta-lactamase-producing strains among H. influenzae isolated in 1994 and 1995 were 20 and 15.8%, respectively, which were slightly higher than those in the past. One quinolone resistant strain was isolated in this study. Although the ratio of beta-lactamase-producing strains among M. catarrhalis was as high (96.7%) as in the past, no increased resistance against the drugs examined was observed.

Anti-Bacterial Agents↗

[Moraxella (Branhamella) catarrhalis: its isolation in the respiratory secretions of adult patients].

OBJECTIVE: We have designed a retrospective study in order to know the clinical significance of the isolation of Moraxella (Branhamella) catarrhalis (MC) in respiratory specimens of adult hospitalized patients. METHODS: We performed a Gram stain and culture on blood-agar, MacConkey media and quantitative culture in chocolate-agar to all respiratory samples. In patients with a clinical diagnosis of pneumonia BCYE-alpha was added. During 2 years (1992-1993) MC was isolated in respiratory specimens from 52 patients. We revised the clinical history of all these patients. RESULTS: MC was isolated in 60 respiratory specimens (sputum and/or tracheobronchial aspirates) from 52 patients. The Gram stain showed gram-negative cocci in 77% and gram-positive cocci in 17% of the cases. MC grew in pure culture in 28 specimens (46.6%). In 23% of cases MC was isolated with Streptococcus pneumoniae and in 21% with Haemophilus influenzae. Fifty-two stocks (86.6%) produced beta-lactamase. Twelve patients had a clinical diagnosis of pneumonia, 8 of them had an underlying chronic respiratory disease. Other 24 patients with an underlying chronic respiratory disease had a bronchial infection as a cause of exacerbation of their respiratory disease. Seven patients without an underlying chronic respiratory disease had a clinical episode of acute bronchitis. Finally, in 9 patients the isolation of MC was considered a colonization. CONCLUSIONS: In 17% cases MC was identified as a gram-positive cocci in the Gram stain, which may cause false diagnosis. The etiological importance of MC in episodes of acute exacerbation of patients with an underlying chronic respiratory disease is high.

Adult↗

Human antibody response to Moraxella catarrhalis antigens.

Moraxella catarrhalis was isolated from 68 of 200 (34%) sputum and 56 (28%) nasopharyngeal swab samples of patients with community-acquired pneumonia. Of the 68 pneumonia patients, 42 (61.8%) were males and 26 (38.2%) females. Fifty one of the 68 patients (75%) had chronic underlying diseases. beta-lactamase was produced by 37 (54.4%) of the 68 sputum samples and 32(57.1%) of the 56 nasopharyngeal isolates. In an ELISA using outer membrane protein antigens of M. catarrhalis against patient sera showed 40 of 68 (58.8%), and 43 of 68 (63.2%) significant increase in convalescent to acute sera when IgA, IgM and IgG3 were used respectively. In control sera only of 30(3.3%) and none showed significant antibody rise when IgA, IgM and IgG3 conjugates were used respectively (P < 0.05).

Acute-Phase Proteins↗

beta-Lactam resistance and beta-lactamase isoforms of Moraxella catarrhalis isolates in Taiwan.

Moraxella catarrhalis is an important pathogen in both upper and lower respiratory tract infections. More than 90% of isolates worldwide produce beta-lactamase. The beta-lactamases produced by M. catarrhalis can be differentiated by isoelectric focusing (IEF) into BRO-1 and BRO-2 patterns. In this study, we investigated the prevalence of various beta-lactamase isoforms in clinical isolates of M. catarrhalis in Taiwan, as well as the relationships of these isoforms with antibiotic resistance. A total of 271 clinical isolates of M. catarrhalis were collected from 12 large medical laboratories in Taiwan from 1 August 1993 to 31 July 1995. The overall prevalence of beta-lactamase production was 98.2% (266 of 271 isolates). Analytical IEF revealed BRO-1 was the most common beta-lactamase pattern among the isolates (238 isolates, 88%); BRO-2 was the only other pattern found, with 32 (12%) isolates. The geometric mean minimum inhibitory concentration of ampicillin for BRO-1 producers was 63-fold higher than that for beta-lactamase-negative isolates, and 6.5-fold higher than that for BRO-2 producers. beta-Lactam antibiotics, such as amoxicillin + clavulanate and the cephalosporins, tested were very active against this species, regardless of whether the isolate produced beta-lactamase or not. In conclusion, beta-lactamase is common among clinical isolates of M. catarrhalis in Taiwan, with BRO-1 being the most common isoform. However, because most isolates tested were still sensitive to amoxicillin + clavulanate and cephalosporins, these agents appear to be reliable alternatives to first-line therapy when M. catarrhalis is contributing to a clinical infection.

Anti-Bacterial Agents↗

Effects of vaccination with a Moraxella bovis bacterin on the subsequent development of signs of corneal disease and infection with M bovis in calves under natural environmental conditions.

A vaccination study was conducted for infectious bovine keratoconjunctivitis (IBK) in 440 purebred Hereford cattle (cows and their newborn calves) of the USDA Meat Animal Research Center cattle herd at Clay Center, Ne. The cattle were allotted to 4 groups: 60 calves were vaccinated with an autogenous Moraxella bovis bacterin (group 1); 60 calves that were matched with group 1 calves were designated nonvaccinated matched controls (group 2); 99 calves were peer group nonvaccinated controls (group 3); and 219 cows, the dams of the calves, were nonvaccinated consorts (group 4). The infection rates in cattle groups 1, 2, 3, and 4 during the summer were 96.6, 98.3, 100, and 79.1%, respectively, and the disease rates were 90, 93, 85, and 20%. The infection and the disease rates were significantly (P less than 0.01) different between claves and cows. The disease rate was also significantly different between older and younger cows. A larger percentage of the affected calves and cows had mild or moderate (61%) signs of IBK rather than severe (39%) signs. The rate of body weight gain was reduced in calves with severe signs of IBK. The results seemed to indicate that little would be gained by vaccinating cattle against IBK under the conditions of study.

Animals↗

Detection of Moraxella bovis antibodies in infectious bovine keratoconjunctivitis by a passive hemagglutination test.

A passive hemagglutination test was developed to detect antibody response to Moraxella bovis in tears. Tannic acid-treated sheep erythrocytes were sensitized with sonicated antigen prepared from M bovis cultures. The test was found to be a relatively simple, specific, and reliable procedure for titrating antibodies in lacrimal secretions. The hemagglutination test could be a valuable method for seroepizootiologic investigation of infectious bovine keratoconjunctivitis.

Animals↗

Diffusion of macrolide antibiotics through the outer membrane of Moraxella catarrhalis.

We reported previously that the high susceptibility of Moraxella catarrhalis to macrolide antibiotics and other hydrophobic antimicrobial agents was related to the hydrophobicity of the cell surface. Electrophoretic analysis of lipopolysaccharide (LPS) extracted from M. catarrhalis revealed a deep rough-type profile similar to that of an LPS Re type mutant of Salmonella typhimurium, which also exhibits high susceptibility to macrolides. Moreover, treatment of 32P-labeled cells of M. catarrhalis by phospholipase C induced the release of radioactive materials. These results suggested that hydrophobic agents such as macrolides readily access the cell surface exposed by the deep rough-type LPS and phospholipids, and permeate into the cell interior through the lipid bilayer. In fact, M. catarrhalis cells rapidly accumulated large amounts of the macrolide antibiotics, erythromycin and rokitamycin, whereas no accumulation of the macrolides was observed in cells having smooth-type or Rc type LPS under the same conditions.

Journal Article↗

The other siblings: respiratory infections caused by Moraxella catarrhalis and Haemophilus influenzae.

Respiratory infections remain substantial causes of morbidity and mortality globally. In this paper, two substantial players in bacterial-associated respiratory disease are assessed as to their respective roles in children and adults and in the developed and developing world. Moraxella catarrhalis, although initially thought to be a nonpathogen, continues to emerge as a cause of upper respiratory disease in children and pneumonia in adults. No vaccine is currently available to prevent M. catarrhalis infection. Haemophilus influenzae type b, originally thought to be the cause of influenza, has now been limited epidemiologically in the developed world due to an effective immunization but it continues to be a major player in the developing world. Nonencapsulated strains of H. influenzae still remain as significant causes of respiratory infections in the developing world especially in exacerbation of chronic obstructive lung disease. Finally, and in brief, the spectrum of Brazilian purpuric fever due to a specific biotype of H. influenzae is discussed.

Journal Article↗

Alpha1-antitrypsin inhibits Moraxella catarrhalis MID protein-induced tonsillar B cell proliferation and IL-6 release.

Alpha1-antitrypsin (AAT) is a major circulating and tissues inhibitor of serine proteinases implicated in the regulation of inflammation and host defence. There is now increasing evidence that AAT may also exhibit anti-inflammatory activities independent of its protease inhibitor function. This study was undertaken to investigate the effects of native (inhibitory) and polymerized (non-inhibitory) forms of AAT on MID (Moraxella IgD binding protein)-induced human tonsillar B cell activation in vitro. We found that 0.5 microg/ml MID induces B cell proliferation and stimulates IL-6 release (p<0.001) relative to non-stimulated controls. Both native and polymerized AAT (0.5 mg/ml) inhibited MID-stimulated B cell proliferation in a similar manner (by 70%, p<0.001), whereas MID-induced IL-6 release was more strongly suppressed by polymerized (9.9-fold, p<0.001) as compared to native AAT (2.8-fold, p<0.01). Electrophoretic analysis of cell culture media did not indicate any interaction between AAT and MID, and flow cytometry data showed no competition for the same receptor. The effects of AATs were observed whether added together with MID or 2h after MID-addition to cell cultures. Thus, our data demonstrate that AAT inhibits MID-induced B cell activation in vitro that is unrelated to its protease inhibitory activity and is not dependent on MID binding to the cell surface.

Adhesins, Bacterial↗

Synthesis of Oligosaccharide Structures from the Lipopolysaccharide of Moraxella catarrhalis.

The synthesis of the octasaccharide [p-(trifluoroacetamido)phenyl]ethyl 4-O-[2-O-(2-acetamido-2-deoxy-alpha-D-glucopyranosyl)-beta-D-glucopyranosyl]-6-O-[2-O-[4-O-(4-O-alpha-D-galactopyranosyl-beta-D-galactopyranosyl)-alpha-D-glucopyranosyl]-beta-D-glucopyranosyl]-3-O-beta-D-glucopyranosyl-alpha-D-glucopyranoside, representing the outer part of the lipooligosaccharide from Moraxella catarrhalis serotype A, is described, together with a hepta-, a hexa-, and a pentasaccaride, composing parts thereof with shorter oligosaccharide chains substituted in the 6-position of the central 3,4,6-branched glucose moiety. The versatility of the use of thioglycosides in oligosaccharide synthesis is shown, since throughout the synthesis thioglycosides are used as glycosyl donor precursors, either directly in dimethyl(methylthio)sulfonium triflate (DMTST)-promoted coupling reactions or after conversion to the corresponding glycosyl bromide in silver triflate-promoted couplings. The effects of different protecting groups, anomeric leaving groups, and solvents used in the various coupling reactions are often substantial, which necessitates the use of easily convertible intermediates.

Journal Article↗

Development and Validation of a Multiplex PCR-Based Assay for the Upper Respiratory Tract Bacterial Pathogens Haemophilus influenzae, Streptococcus pneumoniae, and Moraxella catarrhalis.

Background: Conventional simplex polymerase chain reaction (PCR)-based assays are limited in that they only provide for the detection of a single infectious agent. Many clinical diseases, however, present in a nonspecific, or syndromic, fashion, thereby necessitating the simultaneous assessment of multiple pathogens. Panel-based molecular diagnostic testing can be accomplished by the development of multiplex PCR-based assays, which can detect, individually or severally, different pathogens that are associated with syndromic illness. As part of a larger program of panel development, an assay that can simultaneously detect Haemophilus influenzae, Streptococcus pneumoniae, and Moraxella catarrhalis was developed. These organisms were chosen as they are the most common bacterial pathogens associated with both the acute and chronic forms of otitis media; they are also responsible for a high percentage of sinus infections in both children and adults. In addition, H. influenzae and S. pneumoniae are commonly associated with septic meningitits. Methods and Results: Multiple individual PCR-based assays were developed for each of the three target organisms which were then evaluated for sensitivity and specificity. Utilizing the simplex assays that met our designated performance criteria, a matrix style approach was used to develop a duplex H. influenzae-S. pneumoniae assay. The duplex assay was then used as a single component in the development of a triplex assay, wherein the various M. catarrhalis primer-probe sets were tested for compatibility with the existing assay. A single-step PCR protocol, with species-specific primers for each of the three target organisms and a liquid hybridization-gel retardation amplimer detection system, was developed, which amplifies and then discriminates among each of the amplification products according to size. This assay is able to detect all three organisms in a specific manner, either individually or severally. Dilutional experiments indicate a detection limit of 10 femtograms (fg)(6-7 genomic equivalents) or less of genomic DNA for each of the three microorganisms regardless of the presence of irrelevant DNA. Conclusions: The reliance on individual, robust, species-specific primers and the avoidance of a nested PCR approach make this bacterial multiplex assay suitable for use in the clinical laboratory. This assay has proved useful in both research and patient care applications.

Journal Article↗

The immunoglobulin D-binding part of the outer membrane protein MID from Moraxella catarrhalis comprises 238 amino acids and a tetrameric structure.

Moraxella catarrhalis IgD-binding protein (MID), a 200-kDa outer membrane protein comprising 2,139 amino acids, has recently been isolated and shown to display a unique and specific affinity for human IgD. To identify the IgD-binding region, MID was digested with proteases. In addition, a series of truncated fragments of MID were manufactured and expressed in Escherichia coli followed by analysis for IgD binding in Western and dot blots. The smallest fragment with essentially preserved IgD binding was comprised of 238 amino acid residues (MID(962-1200)). Shorter recombinant proteins gradually lost IgD-binding capacity, and the shortest IgD-binding fragment comprising 157 amino acids (MID(985-1142)) displayed a 1,000-fold reduced IgD binding compared with the full-length molecule. The truncated MID(962-1200) was efficiently attracted to a standard IgD serum and to purified myeloma IgD(kappa) and IgD(lambda) sera but not to IgG, IgM, or IgA myeloma sera. Furthermore, the fragment specifically bound to peripheral blood B lymphocytes, and the binding was inhibited by preincubation with anti-IgD-Fab polyclonal antibodies. Results obtained by introducing five amino acids randomly into MID(962-1200) using transposons suggested that alpha-helix structures were important for IgD binding. Ultracentrifugation experiments and gel electrophoresis revealed that native MID(962-1200) was a tetramer. Interestingly, tetrameric MID(962-1200) attracted IgD more than 20-fold more efficiently than the monomeric form. Thus, a tetrameric structure of MID(962-1200) is crucial for optimal IgD-binding capacity.

Adhesins, Bacterial↗

The in vitro activity of moxifloxacin against Haemophilus influenzae and Moraxella catarrhalis explored using a pharmacodynamic model.

OBJECTIVE: To assess the antibacterial action of moxifloxacin on Haemophilus influenzae and Moraxella catarrhalis using an in vitro pharmacodynamic model of infection. METHODS: Serum concentrations in humans associated with doses of 400 mg once a day for 48 h were simulated and the antibacterial effect measured by the log change in viable count at intervals through the simulation compared to time zero and also the area under the bacterial kill curve (AUBKC). Wild-type strains of H. influenzae and M. catarrhalis with varying beta-lactam susceptibilities (moxifloxacin MICs </=0.1 mg/L) and laboratory-generated mutants with moxifloxacin MICs of 0.5-1.0 mg/L were tested. RESULTS: H. influenzae strains with MICs of </=0.06 mg/L were eradicated by 12 h; strains with MICs of 0.5-1.0 mg/L were eradicated by 24-36 h. M. catarrhalis strains with MICs of </=0.08 mg/L were eradicated by 24-36 h, while the less susceptible laboratory mutant (MIC 1.0 mg/L) was not eradicated even at 48 h. Although we used only a single dosing simulation, the AUC/MIC ratio was strongly related to the AUBKC. CONCLUSIONS: These data indicated that once daily dosing of moxifloxacin holds promise for the treatment of infection due to H. influenzae or M. catarrhalis but bacterial killing is related to MIC.

Journal Article↗

Pathway for Biodegradation of p-Nitrophenol in a Moraxella sp.

A Moraxella strain grew on p-nitrophenol with stoichiometric release of nitrite. During induction of the enzymes for growth on p-nitrophenol, traces of hydroquinone accumulated in the medium. In the presence of 2,2'-dipyridyl, p-nitrophenol was converted stoichiometrically to hydroquinone. Particulate enzymes catalyzed the conversion of p-nitrophenol to hydroquinone in the presence of NADPH and oxygen. Soluble enzymes catalyzed the conversion of hydroquinone to gamma-hydroxymuconic semialdehyde, which was identified by high-performance liquid chromatography (HPLC)-mass spectroscopy. Upon addition of catalytic amounts of NAD, gamma-hydroxymuconic semialdehyde was converted to beta-ketoadipic acid. In the presence of pyruvate and lactic dehydrogenase, substrate amounts of NAD were required and gamma-hydroxymuconic semialdehyde was converted to maleylacetic acid, which was identified by HPLC-mass spectroscopy. Similar results were obtained when the reaction was carried out in the presence of potassium ferricyanide. Extracts prepared from p-nitrophenol-growth cells also contained an enzyme that catalyzed the oxidation of 1,2,4-benzenetriol to maleylacetic acid. The enzyme responsible for the oxidation of 1,2,4-benzenetriol was separated from the enzyme responsible for hydroquinone oxidation by DEAE-cellulose chromatography. The results indicate that the pathway for biodegradation of p-nitrophenol involves the initial removal of the nitro group as nitrite and formation of hydroquinone. 1,4-Benzoquinone, a likely intermediate in the initial reaction, was not detected. Hydroquinone is converted to beta-ketoadipic acid via gamma-hydroxymuconic semialdehyde and maleylacetic acid.

Journal Article↗

[Moraxella catarrhalis: an emerging respiratory pathogen].

In the past years Moraxella (Branhamella) catarrhalis has finally gained respect as a pathogen thanks to the many reports of its causal role. The intent of this review is to provide a critical evaluation of the intent of this review is to provide a critical evaluation of the microbiological features (taxonomy, diagnosis, virulence, epidemiology and drug resistance), clinical diseases and therapy of this microorganism

English Abstract↗

Enhancement of Moraxella bovis-induced keratitis of mice by exposure of the eye to ultraviolet radiation and ragweed extract.

Effects of a ragweed extract and ultraviolet (UV) radiation on Moraxella bovis-induced keratitis of DBA/2J mice were investigated. Mouse eyes were exposed to ragweed extract and/or UV radiation during induction of keratitis. Pathogenesis was enhanced by exposure to ragweed extract alone and by exposure to UV radiation alone. It was enhanced most when eyes were exposed to ragweed extract and UV radiation.

Allergens↗