Microflora of the upper respiratory tract of normal young children and in respiratory tract diseases. Normal microflora and dysbiosis at different levels of the upper respiratory tract in healthy children and pneumonia patients

Material obtained from the mucous membranes of the upper respiratory tract (the anterior section of the nasal cavity, the fauces) in young children, both healthy and suffering with different forms of acute pneumonia, has been analyzed with due regard to the structure of the microflora, its specific composition and the size of populations formed by different species constituting the microflora. This analysis has made it possible to determine the species constituting normal microflora, to detect its dysbiotic changes, and to determine their degree (partial or complete dysbacteriosis). The degree of pathologic changes in the microecological balance of the upper respiratory tract has been shown to reflect the severity of acute pneumonia and to be determined by the characteristics of the natural resistance system in young children.     

Comparative characterization of the microflora of the upper respiratory tract mucous membranes in bronchial asthma and allergic persistent rhinitis

A total of 250 patients with diagnosed bronchial asthma (BA) were examined by microbiological methods. Among them--188 children and 62 adults. In 87 patients the microflora of nasal mucosa was studied, in 40--of pharynx only and in 123 patients--both the nasal and the pharynx. For comparative analysis earlier data obtained in 69 patients with persistent allergic rhinitis (PAR) were used. The cultures isolated from the nasal mucosa of BA patients were shown to number 18 genera and 42 species, while among those isolated from pharynx mucosa 20 genera and 40 species. Monocultures were isolated from the nasal mucosa only in 23% of the examined patients and from the pharynx mucosa--only in 1.42%. Associations with different numbers of components were isolated from nasal and pharynx mucosa (2 to 6, 2 to 8 respectively). Staphylococcus aureus was regarded as the main species of nasal biocenosis in BA and PAR, as well as pharynx biocenosis in BA. Besides, in BA other Staphylococcus species (schleiferi, caprae, capitis, hominis, etc.), reversely related to the main species, could be isolated from both mucous membranes. Similarities and differences in microflora of biocenoses in both nosological forms, confirming links between PAR and BA, are considered.     

Disturbance of immune characteristics of the mucous membranes of the respiratory tract in bronchopulmonary diseases in children

A total of 100 children with bronchopulmonary diseases were examined. Of these, in 80 bronchial asthma and in 20 children acute pneumonia were diagnosed. The deficiency of local cell mediated and humoral factors on the mucous membranes of the upper respiratory tracts was established. In the squamous and columnar epithelium cells an increased destruction and cytolysis was noted leading to disturbances in the integrity of the protective barrier of the epithelial cover. In addition, this study revealed the deficiency of neutrophilic leukocytes, pronounced local eosinophilia and the low level of humoral protective factors. In acute pneumonia an increase in the amount of neutrophilic leukocytes in the secretion of the nasal cavity together with the appearance of destructive processes in epithelial cells took place. Pronounced deficiency in serum IgA and IgG was accompanied by compensatory increase in secretory IgA.     

Intestinal microbiocenosis in children with thymomegaly and acute lower respiratory tract infections and regimen of its correction

57 children with thymomegaly from 3 months to 3 years of age with acute lower respiratory tract infections were studied. Disturbances of gut microflora - changes in both obligate and potentially harmful symbionts were detected in 70.2% of cases. In 47,5% of cases increased quantity of enterococci was observed. Decreased quantities of bifidobacteria and lactobacilli were observed in all and 27.5% of studied patients respectively. Most diverse gut microflora has been observed in children with pneumonia and thymomegaly of II level. During treatment of children with thymomegaly changes in gut microflora should be considered along with changes in the immune system.     

Aetiology of upper respiratory tract infections in children in Arak city: a community based study

Viruses are frequent causes of upper respiratory tract infections in children. We investigated the viral aetiology of community-acquired upper respiratory tract infections (URIs) in young children treated as outpatients in community settings. During November 2008, nasal swab specimens were taken from children with recent onset of upper respiratory tract infections. The patients attended day care or primary schools; the specimens were randomly obtained by pediatricians from schools and childcare institutions and sent for identification by PCR method. A total of 300 specimens were collected. From all samples, 40.67% were positive for at least 1 virus, viz. adenovirus 11.76%, rhinovirus 9.8%, respiratory syncytial virus 6.08%, influenza virus 5.56%, parainfluenza virus 4.9%, enterovirus 2.94% and a combination of 2 viruses 2%. Clinical manifestations of the respiratory infections were as follows: 70.7% of the patients had coryza, 69.3% cough, 26% sneezing, 19.7% sore throat, 2.7% headache, 7.7% fever, 2.3% conjunctivitis, 1.3% abdominal pain and 1% hoarseness. The results of this study demonstrate that adenoviruses and rhinoviruses are the two most common viral agents isolated from pediatric outpatients with acute URIs in autumn in Arak City. Coryza and cough were the most common symptoms in children. Sore throat and hoarseness were more prevalent in infections caused by influenza virus, conjunctivitis in parainfluenza, and coryza in rhinovirus infections.     

Importance of associated microorganisms in the development of acute pneumonia

Sputum samples obtained from 106 patients with acute pneumonia have been studied by the quantitative microbiological method. Different microbial associations have been shown to play an important role in the development of acute pneumonia (67.9%). Microbiological studies have revealed the prevalence of pneumococci in the etiological structure of the disease. Staphylococci, hemophilic bacteria and Neisseria have been found to take part in the development of acute pneumonia in a lesser number of cases and more often occur in combination with pneumococci or form different associations themselves. The so-called "etiological" type of microbial associations has been determined, two or three microbial species being isolated at high concentrations (10(6) and over) and the percentage of patients with such combinations being 59.7 +/- 5.8%. The analysis of the clinical course of the disease has revealed that the presence of different microbial species in patients linked, to a great extent, with the clinical picture of the disease. These data may be used for prescribing adequate etiotropic therapy and for prognostication. Quantitative bacteriological studies carried out in the dynamics of the disease permit timely detection of changes in the microflora of the respiratory tracts, the evaluation of the effectiveness of antibacterial therapy in the process of its implementation and the rational correction of therapeutic measures.     

Autoflora in the upper respiratory tract of Apollo astronauts.

The typical microbial inhabitants of the oral and nasal cavities of Apollo astronauts were identified before space flight and generally found to be similar to those previously reported for healthy male adults. Additional analyses of samples collected immediately after return of the Apollo 13, 14, 15, and 16 crew members to earth were performed to evaluate the effects of space travel on the microbial bioburden of the upper respiratory tract. In-flight cross-contamination and buildup of pathogens such as Staphylococcus aureus were noted, although significant increases in nonpathogenic species were absent. Other proposed alterations, such as dysbacteriosis (flooding of the mouth with a single species) and simplification of the autoflora, did not occur. Generally, the incidence and quantitation of each species after flight was within the preflight range, although the number of viable Haemophilus cells recovered from the mouth decreased significantly after space flight. Except for those minor alterations listed above, the aerobic and anaerobic bacterial components of the upper respiratory autoflora of Apollo astronauts was found to be stable after space flight of up to 295 h.     

Nosopharyngeal microflora in ambulatory treated children and adults with upper respiratory tract infections

Upper respiratory tract consists resident and transient bacterial microflora, which in appropriate condition can cause infection. Bacteriological study was performed among 201 patients with upper respiratory tract infections treated in ambulatory. From nasal and pharyngeal swabs Staphylococcus aureus, Haemophilus influenzae, Streptococcus pneumoniae, Moraxella catarrhalis, and Streptococci group A, B, C, G were isolated. Antibiotic susceptibility testing of isolated strains was performed using CLSI criteria. All isolated strains of streptococci were susceptible to penicillin; some of them demonstrated resistance to macrolides and lincosamides. Few isolated strains of H. influenzae demonstrated resistance to penicillin and cotrimoxazole. Azitromycin resistant strains were not detected. All isolated strains of M. catarrhalis were beta-lactamase positive and demonstrated resistance to penicillin. Strains of methicillin sensitive S. aureus (MSSA) were isolated most frequently from pharyngeal swabs (35.4%) and S. pneumoniae (33.3)--from nasal swabs.     

肺炎支原体和沙眼衣原体致儿童急性下呼吸道感染的研究

本文旨在研究儿童社区获得性急性下呼吸道感染(ALRTI)中肺炎支原体(MP)和沙眼衣原体(CT)的感染特征。采用实时荧光定量聚合酶链反应(PCR)检测2006年10月～2008年2月因ALRTI收入复旦大学附属儿科医院的患儿呼吸道标本中MP和CT的DNA,并分析2种病原体感染患儿的临床特征与实验室检查结果。在1312份深部鼻咽分泌物标本中,MP和CT检出率分别为7.85%(103/1312)和2.97%(39/1312)。MP在5岁以上患儿中的检出率为33.33%(30/90),而CT在3个月以内患儿中的检出率为6.28%(31/494)。MP感染后易出现40℃以上高热,较少发生发绀与重症呼吸道感染;白细胞计数常无明显升高,但C反应蛋白升高较多见。CT感染后40℃以上高热和重症呼吸道感染少见,C反应蛋白升高也较少见。结果提示,在5岁以上儿童的社区获得性ALRTI中,MP是重要的病原体;而在3个月以内儿童中,CT为常见病原体之一。     

冬季肺炎和哮喘急性加重患儿呼吸道微生物多样性

目的研究冬季肺炎和哮喘患儿呼吸道微生物的多样性。方法选择2017年11月至2018年1月在我院急诊科治疗的确诊为肺炎和哮喘急性加重的患儿159例,其中肺炎患儿102例,哮喘急性加重患儿57例。选择同时期在本院就诊的无呼吸道疾病患儿88例,设为对照组。检测患儿呼吸道微生物分布情况。结果肺炎和哮喘患儿呼吸道微生物多样性增加。肺炎患儿菌群丰度前3位的菌属分别是链球菌属、不动杆菌属、克雷伯菌属。哮喘患儿菌群丰度前3位的分别是嗜血杆菌属、莫拉氏菌属、葡萄球菌属。呼吸道病毒检测结果显示,肺炎患儿检出率前3位的病毒分别是肺炎支原体、呼吸道合胞病毒和副流感病毒3型。哮喘患儿病毒检出率前3位的是柯萨奇病毒、呼吸道合胞病毒和肺炎支原体。肺炎患儿肺炎支原体检出率明显高于哮喘患儿,柯萨奇病毒检出率明显低于哮喘患儿。结论冬季肺炎和哮喘患儿的临床表现和体征较为相似,但是病原微生物检出情况有所不同。病原微生物检出的差异性有助于正确诊断和鉴别儿童肺炎和哮喘。     

Dysbiosis of upper respiratory tract microbiota in elderly pneumonia patients

Bacterial pneumonia is a major cause of morbidity and mortality in elderly. We hypothesize that dysbiosis between regular residents of the upper respiratory tract (URT) microbiome, that is balance between commensals and potential pathogens, is involved in pathogen overgrowth and consequently disease. We compared oropharyngeal microbiota of elderly pneumonia patients (n=100) with healthy elderly (n=91) by 16S-rRNA-based sequencing and verified our findings in young adult pneumonia patients (n=27) and young healthy adults (n=187). Microbiota profiles differed significantly between elderly pneumonia patients and healthy elderly (PERMANOVA, P<0.0005). Highly similar differences were observed between microbiota profiles of young adult pneumonia patients and their healthy controls. Clustering resulted in 11 (sub)clusters including 95% (386/405) of samples. We observed three microbiota profiles strongly associated with pneumonia (P<0.05) and either dominated by lactobacilli (n=11), Rothia (n=51) or Streptococcus (pseudo)pneumoniae (n=42). In contrast, three other microbiota clusters (in total n=183) were correlated with health (P<0.05) and were all characterized by more diverse profiles containing higher abundances of especially Prevotella melaninogenica, Veillonella and Leptotrichia. For the remaining clusters (n=99), the association with health or disease was less clear. A decision tree model based on the relative abundance of five bacterial community members in URT microbiota showed high specificity of 95% and sensitivity of 84% (89% and 73%, respectively, after cross-validation) for differentiating pneumonia patients from healthy individuals. These results suggest that pneumonia in elderly and young adults is associated with dysbiosis of the URT microbiome with bacterial overgrowth of single species and absence of distinct anaerobic bacteria. Whether the observed microbiome changes are a cause or a consequence of the development of pneumonia or merely coincide with disease status remains a question for future research.     

High correlation between the bacterial species in denture plaque and pharyngeal microflora

Objectives : The morbidity and mortality of the dependent elderly that result from aspiration pneumonia are recognized as a major geriatric health problem. Most cases of bacterial pneumonia are initiated following colonization or superinfection of the pharynx by pathogenic bacteria, followed by aspiration of pharyngeal contents. A recent study revealed that bacteria, that commonly cause respiratory infection, colonized the dentures of dependent elderly. This suggests that denture plaque may function as a reservoir of potential respiratory pathogens to facilitate colonization on the pharynx. The purpose of this study was to determine the possible correlation between denture and pharyngeal microflora. Study Design : The denture and pharyngeal bacterial flora of 50 dependent elderly were examined, and the microorganisms identified by culturing. The agreement between the bacterial species in denture plaque and pharyngeal microflora was investigated using the Kappa method. Results : The microorganism species on the dentures and pharyngeal mucosa of the subjects had an agreement rate of 68.5%. The agreement rate for each of the bacterial species of the dentures and pharynx was also demonstrated to be high. Conclusions : Dentures should be considered an important reservoir of organisations which could colonise the pharynx, and the importance of controlling denture plaque for the prevention of aspiration pneumonia cannot be overemphasized.     

Determination of pneumococcus antibodies in secretions in acute pneumonia in children

IgA-antibodies to pneumococcal antigens were measured by enzyme immunoassay in saliva and nasal washings, obtained from 32 children aged 6 months to 14 years with acute pneumonia and from 25 healthy children aged 2.5-11 years. In the secretions of children with acute pneumonia an essential increase in the levels of IgA-antibodies in comparison with those observed in healthy children was detected. The levels of antibodies to pneumococci in saliva and in nasal washings were in direct correlation. These data are indicative of a protective role played, probably, by secretory antibodies at the portals of infection in pneumococcal pneumonia.     

Microbiocenosis of the large intestine in acute complicated pneumonia in children

Fifty-six children of early age with pneumonia developed on the background of frequent respiratory infections were placed under observation. In 91.1% of these cases microecological disturbances in the intestine were detected, 46.6% of the patients having third-degree dysbacteriolysis. In such cases it is more correct to regard the "intestinal" syndrome as the clinical manifestation of disturbances in the biocenosis of the intestine, the state of its microflora. Intestinal dysbacteriosis aggravates the course of acute aggravated pneumonia in children and requires the inclusion of special therapy aimed at normalizing intestinal microflora.     

Microbial flora variations in the respiratory tract of mice.

A stable microbial system in the respiratory tract acts as an important defense mechanism against pathogenic microorganisms. Perturbations in this system may allow pathogens to establish. In an ecological environment such as the respiratory tract, there are many diverse factors that play a role in the establishment of the indigenous flora. In the present work we studied the normal microbial flora of different areas of the respiratory tract of mice and their evolution from the time the mice were born. Our interest was to know which were the dominant groups of microorganisms in each area, which were the first capable of colonizing and which dominated over time to be used as probiotic microorganisms. Our results show that Gram negative facultatively anaerobic bacilli and strict anaerobic microorganisms were the last ones to appear in the bronchia, while aerobic and Gram positive cocci were present in all the areas of the respiratory tract. The number of facultative aerobes and strict anaerobes were similar in the nasal passage, pharynx instilled and trachea, but lower in bronchia. The dominant species were Streptococcus viridans and Staphylococcus saprophyticcus, followed by S. epidermidis, Lactobacilli and S. cohnii I which were present on every studied days but at different proportions. This paper is the first part of a research topic investigating the protective effect of the indigenous flora against pathogens using the mice as an experimental model.     

The role of Chlamydia pneumoniae in etiology of acute bronchitis in children

During examination of 60 children aged 3-15 years with manifestations of acute bronchitis Chlamydia pneumoniae antigen was detected in their nasal and pharynx smears in 11.7% of cases with the use of enzyme immunoassay and indirect immunofluorescence test. In 10% of cases chlamydial pneumonia was diagnosed serologically. In the course of complex laboratory diagnostics C. pneumoniae was detected in 13.3% of cases. The most complete information could be obtained by a combination of different methods of the antigen detection as well as specific IgG and IgM antibodies.     

Peripheral blood T cells response in human parainfluenza virus-associated lower respiratory tract infection in children

Human Parainfluenza virus (HPIV) causes lower respiratory tract infections (LRTI) mostly in young children. Respiratory viral infections may decline T cells in circulation and display enhanced pathogenicity. This study is aimed to analyze T cells alterations due to HPIV in children with LRTIs. Children (N = 152) with bronchitis or pneumonia, admitted in tertiary care hospitals were included in the study. Respiratory samples (throat or nasopharyngeal swabs) were taken and HPIV genotypes (1–4) were analyzed through RT-PCR. Peripheral blood T cells, CD3+, CD4+, CD8+, and CD19+, were analyzed in confirmed HPIV positive and healthy control group children through flow cytometry. The positivity rate of HPIV was 24.34% and the most prevalent genotype was HPIV-3 (20.40%). HPIV-1 and HPIV-2 were detected in 0.66% and 02% children respectively. The T lymphocyte counts were observed significantly reduced in children infected with HPIV-3. CD4+ cell (1580 ± 97.87) counts did not change significantly but the lowest CD8+ T cell counts (518.5 ± 74.00) were recorded. Similarly, CD3+ and CD19 cell ratios were also reduced. The CD4/CD8 ratio was significantly higher (3.12 ± 0.59) in the study population as compared to the control group (2.18 ± 0.654). Changes in the count of CD8+ T cells were more pronounced in patients with bronchiolitis and pneumonia. It is concluded that CD8+ T cells show a reduced response to HPIV-3 in children with severe LRTIs suggesting a strong association of these cells with disease severity.     

Microbial incidence in upper respiratory tracts of workers in the paper industry.

The incidence of microbes in the nasal cavities of workers in three paper and board mills was investigated. A total of 234 persons exposed to microbial aerosols and splashes from paper machine wires and debarker drums formed the exposed group. The control group consisted of 294 workers from the dry working areas: the winding and packing sections. Chi-square analysis was used to test the differences in the frequency of microbial incidence and various symptoms between the exposed and control groups. The nasal cavities of many workers, particularly workers in the debarkers, proved to be contaminated by Klebsiella pneumoniae, other coliforms, yeasts, and molds; usually only one microbe was involved, but sometimes two or several species were found. Nasal bacteria and yeasts were largely derived from the mill and debarker air; the microbes in the air came mainly from process waters. Lack of association of nasopharyngeal symptoms with either exposure to aerosols or nasal microbial contamination was interpreted as an indication of host defenses that were adequate to protect workers from harmful microbial colonization in paper mill environments.     

Antibiotic sensitivity of the bacterial microflora from the sputum of children with acute pneumonia]

Bacteriological analysis of sputum of 598 children with acute pneumonia was performed. Sensivity of 1348 cultures belonging to 8 bacterial species with respect to benzylpenicillin, streptomycin, chloramphenicol, tetracycline, oxytetracycline, chlortetracycline, erythromycin, oleandomycin, neomycin and monomycin was determined. It was shown that the sputum microflora was often resistant to the antibiotics widely used in the medical practice for prolonged periods of time, such as benzylpenicillin, streptomycin, chloramphenicol, tetracyclines. However, it usually remained sunsitive to neomycin, monomycin, erythromycin and oleandomycin. It was found that antibioticogrammes defining the antibiotic choice were of great significance for therapy of acute pneumonia.     

健康儿童与轮状病毒感染儿童肠道菌群结构的比较研究

目的比较分析轮状病毒感染个体与健康个体肠道菌群结构的差异。方法采集11例轮状病毒感染个体及6例健康个体的粪便样品,提取粪便样品中细菌的混合DNA,先通过ERIC-PCR结合分子杂交的技术分析两组个体之间肠道微生物组成的相似性;再扩增粪便样品中菌群的16SrRNA基因,利用PCR—TGGE技术分析肠道菌群的组成情况。结果轮状病毒感染个体与健康个体相比,肠道菌群中GC含量较低的菌明显减少,同时肠道菌群有宿主专一性。结论轮状病毒感染会导致儿童肠道内菌群结构失调。