Branhamella catarrhalis Pneumonia

The diagnosis of Branhamella catarrhalis pneumonia in five cases was established by culture of pulmonary secretions obtained by transtracheal aspiration. B catarrhalis caused an acute lobar pneumonia which usually responded promptly to appropriate antimicrobial therapy. Recognition that this organism may cause pneumonia in a nonimmunocompromised person should alert clinicians to consider it as a possible pathogen when Gramnegative diplococci are seen on smears of specimens from the lower respiratory tract.     

Bronchopulmonary infection due to Branhamella catarrhalis.

Over six months Branhamella catarrhalis was isolated in pure culture from the sputum of 81 patients with symptoms of acute respiratory tract infection. Of 38 patients who were infected in the community, over half required admission to hospital. The remaining 43 patients acquired the infection in hospital. Forty one of the 81 isolates produced beta-lactamase, 24 of these being hospital acquired infections. As a result 40% of patients who were treated with ampicillin did not respond. Most patients had chronic lung diseases or lung cancer or were taking corticosteroids. Three patients died and one required assisted ventilation; strains producing beta-lactamase were isolated in each case. Acute bronchitis developed in one previously healthy young non-smoker. It is concluded that B catarrhalis is an important pathogen of the lower respiratory tract which should be reported, and strains producing beta-lactamase should be identified. Otherwise, treatment with inappropriate antibiotics may result in increased morbidity or mortality.     

Electron microscopic observation of Branhamella catarrhalis

The hemagglutination (HA) test was done on 85 strains of Branhamella catarrhalis, isolated from sputum of patients with respiratory infections; 53% were HA positive strains. Three HA positive and three HA negative strains were selected and were observed under the electron microscope. The bacterial cell wall appeared to be lobulated and its total thickness was about 38 nm. The nuclear region consisted of whorls or fibrils and dense bodies. Five strains were fimbriated and one strain was nonfimbriated. The size of fimbriae was about 68 nm in length and 4.5 nm in width. The fimbriae of Branhamella catarrhalis were densely arranged and peritrichous in distribution. There was no change of fimbriation between broth and agar cultures.     

A note on susceptibility of Branhamella catarrhalis to heavy metals

The susceptibility of 56 strains of Branhamella catarrhalis and ten Neisseria spp. to arsenate, silver, nickel, mercury, lead, cadmium, chromium, manganese, iron, cobalt and molybdenum was tested with an agar dilution technique. All but two strains of B. catarrhalis were resistant to multiple metal ions. There were not sufficient differences in susceptibility, however, to allow the development of a typing scheme based on resistograms. Heavy metal resistance in Branhamella was unrelated to beta-lactamase production. Neisseria spp. were more susceptible to metal ions than B. catarrhalis and this may form the basis of a simple diagnostic test.     

A note on susceptibility of Branhamella catarrhalis to heavy metals

The susceptibility of 56 strains of Branhamella catarrhalis and ten Neisseria spp. to arsenate, silver, nickel, mercury, lead, cadmium, chromium, manganese, iron, cobalt and molybdenum was tested with an agar dilution technique. All but two strains of B. catarrhalis were resistant to multiple metal ions. There were not sufficient differences in susceptibility, however, to allow the development of a typing scheme based on resistograms. Heavy metal resistance in Branhamella was unrelated to beta-lactamase production. Neisseria spp. were more susceptible to metal ions than B. catarrhalis and this may form the basis of a simple diagnostic test.     

Antibacterial activity of cefpodoxime against Branhamella catarrhalis.

The antibacterial activity of cefpodoxime against Branhamella catarrhalis was studied. All of the 65 clinical isolates tested were inhibited at and below 1.56 micrograms/ml, both at 10(7) and at 10(5) CFUs. The following was further studied on B. catarrhalis N-5 which showed average susceptibility to each drug examined. Bactericidal activity was observed at and above the MIC. Scanning and transmission electron microscopy revealed morphological changes, such as cellular swelling, bleb formation, inhibition of septum formation, and lysis, of the cells exposed to cefpodoxime at concentrations around the MIC. Cefpodoxime was poorly hydrolyzed by the beta-lactamase and it showed affinity for two penicillin-binding proteins that had approximate molecular weights of 83 and 74 kilodaltons, with I50 values of 3.7 and 2.1 micrograms/ml, respectively.     

Lipids of Branhamella catarrhalis and Neisseria gonorrhoeae.

Three strains of Branhamella catarrhalis and three strains of Neisseria gonorrhoeae were analyzed with regard to their phospholipid and neutral lipid composition. B. catarrhalis (ATCC 23246) contained 5.12 +/- 0.34% lipid, determined gravimetrically, compared to 8.56 +/- 0.15% and 9.73 +/- 0.06% for two strains of N. gonorrhoeae. Cardiolipin, phosphatidylglycerol, and phosphatidyl-ethanolamine were identified in extracts of both species. In addition, B. catarrhalis contained small amounts of phosphatidylcholine, and N. gonorrhoeae contained small amounts of lyso-phosphatidylethanolamine, which accumulated with autolysis accompanying late cell culture growth. The kinetics of change of relative amounts of phospholipids in both species were measured and found to differ substantially. Neutral lipid accounted for 30.4% of the total lipid of B. catarrhalis (ATCC 23246) and 7.6% of the total lipid of N. gonorrhoeae NYH 002. Hydrocarbons, triglycerides, free fatty acids, coenzyme Q, diglycerides, and free hydroxy fatty acids were identified in the neutral lipid fraction of both species. The three strains of N. gonorrhoeae, sensitive, intermediate, and resistant to penicillin, exhibited no significant difference in the composition or metabolism of phospholipid.     

Possible presence of a capsule in Branhamella catarrhalis.

Clinical isolates of Branhamella catarrhalis from patients with respiratory infections were used in this study. Electron microscopic observation after treating Branhamella catarrhalis with immune serum and ruthenium red revealed the capsule. In the phagocytosis test, most organisms were not ingested by human polymorphonuclear neutrophils in the presence of normal rabbit serum (NRS), while organisms were primarily cell associated and apparently ingested in the presence of immunized rabbit serum (IRS). The capsule may be one of the virulence factors in this bacteria. This study demonstrates the possible presence of a capsule in Branhamella catarrhalis.     

Supersusceptibility to hydrophobic antimicrobial agents and cell surface hydrophobicity in Branhamella catarrhalis

To clarify the cause of the supersusceptibility of Branhamella catarrhalis to macrolide antibiotics, which are well-known to be inactive to most Gram-negative bacteria, we determined its cell surface hydrophobicity by the partition experiment between water and hydrocarbons. Its cell surface was found to be markedly more hydrophobic than that of Escherichia coli or Pseudomonas aeruginosa cells. This suggested that the outer membrane of B. catarrhalis plays no role as a diffusion barrier towards hydrophobic agents.     

一株卡他布朗汉姆菌的重新鉴定

目的对中国医学细菌保藏管理中心库藏的一株卡他布朗汉姆菌CMCC(B)29103株进行重新鉴定。方法用营养琼脂培养基培养CMCC(B)29103株,对其进行形态观察、生理生化特性、脂肪酸组分、分子生物学等多相鉴定,同时与模式株DSM25388T相对照;分析CMCC(B)29103株的特征属性、进化位置以及与Acinetobacter indicus模式株DSM25388T的同源性。结果形态学特性、生理生化以及脂肪酸组分构成均与DSM25388T株十分相似,仅存在个别差异;16 S rRNA基因序列比对显示,CMCC(B)29103株与Acinetobacter(不动杆菌属)相近,与模式株DSM25388T相似性最高,为99.85%。基于Acinetobacter属所有成员的16 S rRNA和rpo B基因的系统进化分析均显示CMCC(B)29103与DSM25388T稳定聚类成一个独立分支,且二者的DNA-DNA同源性为78.3%。结论CMCC(B)29103株属于Acinetobacter indicus种,与模式株DSM25388T为不同的菌株,可将其更名为Acinetobacter i ndicus。     

Characteristics of nonpathogenic Neisseria and Branhamella catarrhalis based on cytopathogenicity

The study of the action of 12 Neisseria species belonging to 112 strains, 6 B. catarrhalis strains and 202 meningococcal strains on the culture of continuous cell line F1 (human amniotic cells) has revealed that nonpathogenic Neisseria are essentially weaker than meningococci in their pathogenicity (expressed in terms of CPD50). Among nonpathogenic Neisseria highly cytopathogenic strains occur in 13.9% of cases, which gives grounds for considering them opportunistic bacteria. Sharply pronounced correlation between the adhesive and pathogenic properties of Neisseria has been observed. The cytopathogenic action of Neisseria is accompanied by the lesion of the chromosomal apparatus of mitotic infected cells.     

Ultrastructural study on the adherence of Branhamella catarrhalis to oropharyngeal epithelial cell.

In the present study, it was observed that Branhamella catarrhalis adhere to the microplicae of the oropharyngeal epithelial cells. Both long and short microplicae patterns are present on the surface of oropharyngeal epithelial cells and the adherence ability of fimbriated Branhamella catarrhalis also varies according to the microplicae pattern. It was found that Branhamella catarrhalis attached more to one surface of the epithelial cell than to the other, suggesting that the presence of receptors are more on one surface than on the other. Branhamella catarrhalis did not attach to the mucus layer but directly to the epithelial cell surface. Ruthenium red staining specimen showed that Branhamella catarrhalis attached to a granular ruthenium red positive layer on the microplicae and also to a ruthenium red positive component, external to the unit membrane of the epithelial cell membrane.     

Transmission electron microscopy studies of Moraxella (Branhamella) catarrhalis

A trypsin-sensitive 200-kDa protein has been reported to be exclusively associated with haemagglutinating isolates of Moraxella (Branhamella) catarrhalis. Transmission electron microscopy studies revealed that haemagglutination by M. catarrhalis to both human and rabbit erythrocytes was mediated by a trypsin-sensitive outer fibrillar coat. This fibrillar layer was absent on non-haemagglutinating isolates examined. Immuno-electron microscopy, using a polyclonal antiserum containing antibodies to the 200-kDa protein as a probe, showed that the 200-kDa protein is present on the outer fibrillar layer of the bacterium. These findings suggest that the haemagglutinin of M. catarrhalis is a 200-kDa protein present on the outer fibrillar coat.     

Expression of Fimbriae and Host Response in Branhamella catarrhalis Respiratory Infections

Sputum during the acute exacerbation of chronic respiratory diseases were observed under the electron microscope, to determine the in vivo expression of surface structures of Branhamella catarrhalis (B. catarrhalis), the polymorphonuclear neutrophil (PMN) response to B. catarrhalis infections, and the composition of sputum. It was found that during infection fimbriae are expressed in B. catarrhalis. However, there were sparsely to densely fimbriated bacteria in each sputum sample. The length of the fimbriae were from 50 to 76 nm. In the sparsely fimbriated B. catarrhalis, external to the cell wall, a thin, granular, electron-dense layer was observed. Due to the presence of fimbriae, this layer was not seen in densely fimbriated B. catarrhalis. Blebs were also found in B. catarrhalis. PMNs were found to phagocytose both B. catarrhalis and debris. Evidence was found that debris were formed mainly by the destruction of PMNs. Bacteria as well as debris were phagocytosed by PMNs.     

Studies on the cellular and free lipopolysaccharides from Branhamella catarrhalis.

Cellular and free lipopolysaccharides obtained from Neisseria catarrhalis and Branhamella catarrhalis were found to be essentially identical. Both cellular and free lipopolysaccharides contained core-oligosaccharides of the following composition: D-glucose (4 mol), D-galactose (1 mol), 2-amino-2-deoxy-D-glucose (1 mol), and 3-deoxy-D-manno-octulosonic acid. Aldoheptose and phosphate components were below levels of detection. Several physical methods indicated that all core-oligosaccharide preparations were identical. Lipid A preparations from cellular and free lipopolysaccharides of both organisms were qualitatively and quantitatively similar; they were composed of decanoic acid, dodecanoic acid, 3-hydroxy dodecanoic acid, 2-amino-2-deoxy-D-glucose, phosphate, and ethanolamine. The results tend to justify the transfer of Neisseria catarrhalis to the genus Branhamella.     

Progress toward the development of a vaccine to prevent Moraxella (Branhamella) catarrhalis infections

Moraxella catarrhalis is a major cause of otitis media and respiratory disease. Vaccine development is at the antigen identification stage. This review examines the more promising antigens, including the 200K protein, the hemagglutinins, the lactoferrin-binding proteins, the UspA proteins, the CopB protein, the transferrin-binding proteins, the CD protein, the E protein and lipooligosaccharide conjugates. Clinical testing of some of these antigens should begin soon.