Early detection and rapid identification of Streptococcus pneumoniae and Haemophilus influenzae in acute pneumonias

The comparative study of the diagnostic value of the enzyme immunoassay (EIA), indirect immunofluorescence (IF) and countercurrent immunoelectrophoresis (CIE) was made. The serological identification of the isolated and reference pneumococci (19) and H. influenzae (38) strains revealed the possibility of using all three microanalytical methods for this purpose. The study of pneumococcal and H. influenzae antigens in native sputum obtained from 74 patients with acute pneumonia showed that EIA and indirect IF were highly sensitive, their sensitivity considerably exceeding that of the bacteriological analysis. Pneumococcal antigens were detected in 66.2% of patients by EIA and in 54.0% of patients by indirect IF, while H. influenzae antigens were detected in 58.1% of patients by EIA and in 67.6% of patients by indirect IF. The sensitivity of CIE proved to be considerably lower; in the detection of pneumococcal antigens it was level with the sensitivity of the bacteriological analysis (23.0%) and H. influenzae antigens could be detected only in 27.0% of patients.     

Haemophilus influenzae biovars in patients with acute and chronic inflammatory lung diseases

In the study of the biological properties of 175 H. influenzae strains isolated from patients with acute and chronic inflammatory lung diseases (ILD) and from donors, a wide spread of biovars I, II and III (according to Kilian) was revealed; these biovars constituted 80% of the cultures under study. In donors, H. influenzae strains were characterized by a wide spectrum of biovars, but biovars I, II and VI constituted more than a half (64.2%) of the strains obtained in the course of these investigations. In acute ILD, only H. influenzae biovars I, II and III were isolated with the prevalence of biovar II (56.4%). In chronic ILD, all H. influenzae biovars were represented, but biovars II and III prevailed (58.7%). The four-fold difference in the occurrence of H. influenzae strains belonging to undetermined biovars was established in donors in comparison with ILD patients (46.7 +/- 9.8% and 12.0 +/- 2.5%; p less than 0.001).     

Nontypeable Haemophilus influenzae inhibits autolysis and fratricide of Streptococcus pneumoniae in vitro

Streptococcus pneumoniae (SP) and nontypeable Haemophilus influenzae (NTHi) are common commensals of the human airway and major bacterial pathogens of otitis media (OM) and other upper airway infections. The interaction between them may play an important role in the pathogenesis of polymicrobial infections. Although previous studies suggested NTHi could promote pneumococcal survival and biofilm formation, how NTHi affects pneumococcal activities has not been defined. Our data in the present studies indicated that the outcome of the interaction between SP and NTHi was in a cell-density-dependent manner and the enhancement of pneumococcal survival happened at the later stages of culturing. Using quantitative PCR, we found that the expression of pneumococcal genes regulating autolysis and fratricide, lytA and cbpD, were significantly down-regulated in co-culture with NTHi. We further observed that influence of NTHi was not on direct cell-to-cell contact, but that this contact may contribute to the interaction between these two microorganisms. These results suggest that pneumococcal survival and biofilm formation can be enhanced by down-regulating pneumococcal cell wall hydrolase production thereby inhibiting pneumococcal autolysis and fratricide in the presence of NTHi.     

Colonization and turnover of Streptococcus pneumoniae, Haemophilus influenzae, and Moraxella catarrhalis in otitis-prone children

Recurrent otitis media are frequently intractable during childhood. It is unclear whether recurrent otitis media is caused by etiological bacteria colonization or by new infections. Streptococcus pneumoniae, Haemophilus influenzae and Moraxella catarrhalis were isolated from the nasopharynx of 7 otitisprone and 2 non-prone children with recurrent otitis media. Plural bacterial species and strains were found in all children while affected by otitis media. The same strain was repeatedly isolated from all otitisprone children even after administration of antibiotics but was not from the non-prone children. Antibiotic susceptibility did not differ significantly among the same repeatedly isolated strains. This pilot study suggests that the etiological bacteria tend to colonize and is hard to eliminate in otitis-prone children.     

Comparative characteristics of the Haemophilus influenzae strains isolated from healthy children and from patients with acute and chronic bronchopulmonary diseases

The comparative biochemical and serological characterization of 424 H. influenzae strains isolated from healthy children and patients with acute and chronic bronchopulmonary diseases is presented. As the result of biotyping H. influenzae strains, 82.3-90.9% of the strains isolated from both healthy children and patients with acute and chronic bronchopulmonary diseases were found to belong to the first three biotypes according to M. Kilian's classification. Among H. influenzae strains isolated from healthy children no capsular variants were detected in the coagglutination test. From patients with acute and chronic diseases of respiratory organs, as a rule, the capsular variants of H. influenzae were isolated (94.4% and 98.1%, respectively). In patients with chronic pneumonia biotypes I, II and III, more seldom biotype V, proved to be mo st invasive. In the determination of the minimum inhibiting concentration of ampicillin, no H. influenzae strains resistant to this antibiotic were detected.     

A comparative study of preservation and storage of Haemophilus influenzae

The aim of this study was to compare the efficacy of conservation by freezing the strains of Haemophilus influenzae at -20 degrees C and -70 degrees C. Skim milk supplemented with glucose, yeast extract and glycerol allowed highest viability of H. influenzae both at -20 degrees C and -70 degrees C from the media analyzed. Trypticase soy broth and brain heart infusion broth supplemented with glycerol, allowed excellent recovery. Use of cotton swaps as supporting material, with or without addition of cryoprotective agents, did not modify H. influenzae viability after six months of storage. Concentration of the initial inoculum positively affected viability when stored at -20 degrees C. Initial concentration did not influence survival after storage at -70 degrees C. Thawing at room temperature should not exceed 3 h as to get highest survival percentage.     

Sequence variation in the hpd gene of nonencapsulated Haemophilus influenzae isolated from patients with chronic bronchitis

The molecular diversity of protein D of nonencapsulated Haemophilus influenzae strains isolated from persistently infected patients with chronic bronchitis was studied by sequencing the hpd gene of four independently obtained isolates. The nucleotide (nt) sequences of the hpd genes of two strains were identical. The other two hpd sequences showed nt substitutions which were mostly synonymous. As a consequence the deduced amino acid (aa) sequences differed from the consensus sequence only by a few aa. No changes in the hpd genes were observed among the four variants of the four strains persisting in chronic bronchitis patients for 9, 11, 8 and 3 months, respectively, although variation in their major outer membrane proteins P2 and P5 occurred. We conclude that the hpd gene is conserved during chronic infections of nonencapsulated H. influenzae.     

Isolation of spheroplastic forms of Haemophilus influenzae from sputum in conventionally treated chronic bronchial sepsis using selective medium supplemented with N-acetyl-D-glucosamine: possible reservoir for re-emergence of infection.

The isolation rate of Haemophilus influenzae from patients with persistent production of purulent sputum has been increased by the routine use of a selective medium. Nevertheless, some purulent sputum still fails to yield a pathogen. The selective medium was supplemented with N-acetyl-D-glucosamine to encourage primary isolation of colony forming, spheroplastic H influenzae, which reverted to normal forms on subculture. On the basis of in vitro experiments it is postulated that these spheroplastic forms of H influenzae may be induced by inadequate antimicrobial chemotherapy and may be responsible for re-emergence of symptoms in these patients during or shortly after stopping chemotherapy.     

儿童肺炎链球菌和流感嗜血杆菌耐药及感染特征分析

目的 了解儿童呼吸道感染肺炎链球菌(Streptococcus pneumonia,SP)和流感嗜血杆菌(Haemophilus influenzae,Hi)的分布特征、耐药情况,及耐药菌抗生素间的相互影响,以更合理地指导临床用药.方法 对2009-2010年临床呼吸道感染患儿进行痰、咽拭子或肺泡灌洗液培养分离Hi和SP.因子需求试验鉴定Hi,头孢硝噻酚法检测β-内酰胺酶;奥普托辛和胆汁溶菌试验确认SP.两菌均采用K-B法检测常用对抗生素的耐药性.结果 收集SP 495株,Hi 515株,多见于3岁以下儿童,以呼吸科分离率最高.SP对红霉素、四环素、阿奇霉素、复方新诺明的耐药率分别为98.4％、66.1％、98％和81.6％,其对青霉素敏感性仅为9.5％.Hi有42.7％产生β-内酰胺酶,Hi对氯霉素、复方新诺明、氨苄青霉素、阿奇霉素的耐药率分别为22.1％、21.6％、36.7％和62.7％.与青霉素敏感SP和β-内酰胺酶阴性Hi相比,耐药SP和阳性Hi更易对氯霉素、四环素和复方新诺明耐药.结论 SP和Hi以婴幼儿为主,多见于呼吸科.其耐药情况严峻,青霉素耐药和产β-内酰胺酶菌株会诱导其他抗生素耐药,引发多重耐药.合理使用抗生素以及对两菌的耐药监测应引起高度重视.     

Cytological and biochemical indices of induced sputum in patients with bronchial asthma and chronic obstructive bronchitis

The aim of our research was to study the apoptotic index and the representativeness of CD95-receptor in the effector cells, in addition to estimation of the content of tumor necrosis factor-alpha in the induced sputum supernatant observed in patients with bronchial asthma and chronic obstructive bronchitis. These indices were evaluated in the disease dynamics and involves various treatment programs. The object of our research was induced sputum. The analysis has proven that apoptotic index of effector cell nuclei, representativeness of CD95-receptor and the content of tumor necrosis factor-alpha in the induced sputum differed, respectively, in patients with bronchial asthma and chronical obstructive bronchitis, and depended on the period of disease and employed treatment. The induced sputum analysis results can be used for the airway inflammation activity monitoring, with regard to these diseases and to treatment efficiency monitoring.     

Culture and morphological characteristics of Haemophilus influenzae and pneumococcus in bronchial infection]

In the cultures obtained by inoculating sputum samples faken from patients with bronchial infection into solid agar medium prepared on Hottinger's hydrolysate with fresh rabbit blood added Haemophilus influenzae produced colonies varying in their from (dome-shaped, conical, trapeziform), as well as in the morphology of the organisms. Pneumococci produced mainly flat colonies surrounded by the zone of alpha hemolysis. Along-side with isolated H. influenzae and pneumococcal colonies, symbiotic colonies could be observed. In these colonies pneumococci were diffused among H. influenzae.     

Bronchial brushing during fiberoptic bronchoscopy for the cytodiagnosis of lung cancer: comparison with sputum and bronchial washings.

In a study of 50 cases of bronchogenic carcinoma in which brushings and washings during fiberoptic bronchoscopy, as well as sputum cytopathologic examinations were performed in the same patients, accuracy rates were respectively: 76 per cent, 76 per cent and 56 per cent. The main cytologic differences setting brush apart from wash and sputum specimens referred to the arrangement of tumor cells as well as the distribution of chromatin within their nuclei. These differences appeared related to cell degeneration which was minimal in brush materials and maximum in sputum specimens. Only six cases were assigned a different cell type of bronchogenic carcinoma when brush cytopathologic diagnoses were compared with results obtained by biopsy or lobectomy specimens. Our findings are consistent with the view that the brush technique is very accurate for the cytodiagnosis of lung cancer and becomes also rather specific once cytologic characteristics of the fresher samples obtained become familiar to the cytopathologist. Non-observance of the special characteristics of these better preserved cellular samples is the major pitfall as to diagnosing, cell typing and judging degree of differentiation of bronchogenic carcinoma in brush cytology specimens.     

Antibiotic sensitivity of Streptococcus pneumoniae strains isolated from the bronchial contents of patients with inflammatory lung diseases

The study of the sensitivity of 2681 pneumococcal strains to antibiotics, carried out in 1976-1982, demonstrated that most of these strains circulating in Leningrad were sensitive to penicillin, levomycetin, erythromycin and oleandomycin. Penicillin-resistant pneumococcal strains were found to appear in 1980 and later showed a definite increase in number, reaching 4.4% of all strains in 1982. Simultaneously the appearance of levomycetin- and erythromycin-resistant strains and the increase of their relative significance were established. Most of the pneumococcal strains isolated in Leningrad were not sensitive to monomycin, streptomycin and tetracyclin.     

A Comparison of a New Oral Streptogramin XRP 2868 with Quinupristin-Dalfopristin Against Antibiotic-Resistant Strains of Haemophilus influenzae, Staphylococcus aureus, and Streptococcus pneumoniae

A new streptogramin antibiotic XRP 2868 was compared with quinupristin-dalfopristin for inhibitory activities against antibiotic-resistant Haemophilus influenzae, Staphylococcus aureus, and Streptococcus pneumoniae. In each organism examined, XRP 2868 had an IC₅₀ that was twofold to fivefold lower than quinupristin-dalfopristin, for inhibition of cell viability, protein synthesis, and ribosomal subunit formation.