PURPOSE: Different characteristics of airway microbiome in asthmatics may lead to differential immune responses, which in turn cause eosinophilic or neutrophilic airway inflammation. However, the relationships among these factors have yet to be fully elucidated.METHODS: Microbes in induced sputum samples were subjected to sequence analysis of 16S rRNA. Airway inflammatory phenotypes were defined as neutrophils (>60%) and eosinophils (>3%), and inflammation endotypes were defined by levels of T helper (Th) 1 (interferon-γ), Th2 (interleukin [IL]-5 and IL-13), Th-17 (IL-17), and innate Th2 (IL-25, IL-33, and thymic stromal lymphopoietin) cytokines, inflammasomes (IL-1β), epithelial activation markers (granulocyte-macrophage colony-stimulating factor and IL-8), and Inflammation (IL-6 and tumor necrosis factor-α) cytokines in sputum supernatants was assessed by enzyme-linked immunosorbent assay.RESULTS: The numbers of operational taxonomic units were significantly higher in the mixed (n = 21) and neutrophilic (n = 23) inflammation groups than in the paucigranulocytic inflammation group (n = 19; p < 0.05). At the species level, Granulicatella adiacens, Streptococcus parasanguinis, Streptococcus pneumoniae, Veillonella rogosae, Haemophilus parainfluenzae, and Neisseria perflava levels were significantly higher in the eosinophilic inflammation group (n = 20), whereas JYGU_s levels were significantly higher in the neutrophilic inflammation group compared to the other subtypes (P < 0.05). Additionally, IL-5 and IL-13 concentrations were correlated with the percentage of eosinophils (P < 0.05) and IL-13 levels were positively correlated with the read counts of Porphyromonas pasteri and V. rogosae (P < 0.05). IL-1β concentrations were correlated with the percentage of neutrophils (P < 0.05). had a tendency to be positively correlated with the read count of JYGU_s (P = 0.095), and was negatively correlated with that of S. pneumoniae (P < 0.05).CONCLUSIONS: Difference of microbial patterns in airways may induce distinctive endotypes of asthma, which is responsible for the neutrophilic or eosinophilic inflammation in asthma.
Asthma ; Colony-Stimulating Factors ; Cytokines ; Enzyme-Linked Immunosorbent Assay ; Eosinophils ; Haemophilus parainfluenzae ; Inflammasomes ; Inflammation ; Interleukin-13 ; Interleukin-33 ; Interleukin-5 ; Microbiota ; Necrosis ; Neisseria ; Neutrophils ; Phenotype ; Pneumonia ; Porphyromonas ; Sequence Analysis ; Sputum ; Streptococcus ; Streptococcus pneumoniae ; Veillonella

The aim of this study was to identify the non-Aggregatibacter actinomycetemcomitans bacteria grown on the tryptic soy-serum-bacitracin-vancomycin (TSBV) medium, an A. actinomycetemcomitans selective medium. A total of 82 unidentified bacterial isolates from the oral cavities of a Korean population were kindly provide by the Korean Collection for Oral Microbiology. All the clinical isolates were grown on TSBV medium and bacterial DNA purified from each isolate was subjected to PCR with universal primers specific for bacterial 16S rRNA genes (16S rDNAs) sequence. The each bacterial 16S rDNA was amplified by PCR and the nucleotide sequences of it was determined by the dideoxynucleotide chain termination method. They were identified by 16S rDNA sequence comparison method at the specie-level. The data showed that Neisseria spp. (42 strains), Fusobacterium spp. (10 strains), Capnocytophaga spp. (8 strains), Propionibacterium acnes (5 strains), Aggregatibacter aprophilus (4 strains), Campylobacter spp. (5 strains), Veillonella dispar (3 strains), Streptococcus sp. (1 strain), Haemophilus parainfluenzae (1 strain), Leptotrichia wadei (1 strain), Morococcus sp./Neisseria sp. (1 strain), and Staphylococcus sp. (1 strain) were identified. These results could be used to develop a new A. actinomycetemcomitans-selective medium which is more effective than the TSBV medium in future studies.
Aggregatibacter ; Bacteria* ; Base Sequence ; Campylobacter ; Capnocytophaga ; DNA, Bacterial ; DNA, Ribosomal ; Fusobacterium ; Genes, rRNA ; Haemophilus parainfluenzae ; Leptotrichia ; Methods ; Neisseria ; Polymerase Chain Reaction ; Propionibacterium acnes ; Staphylococcus ; Streptococcus ; Veillonella

OBJECTIVETo investigate the distribution of pathogens of children with community acquired pneumonia (CAP) from the Chongqing area.

METHODSNasopharyngeal specimens and blood specimens of 1 613 children with CAP were collected between January 2014 and December 2014 for bacterial culture and detection of 7 respiratory viruses and antibodies against Mycoplasma pneumoniae (MP).

RESULTSThe overall positive rate of bacteria was 50.22% (810 cases). Hemophilus parainfluenzae (40.8%), Streptococcus pneumonia (29.7%) and Moraxelle catarrhalis (7.3%) were the predominant ones. Among the viruses, the top detected virus was respiratory syncytial virus (RSV, 58.3%), followed by parainfluenza virus type3 (17.4%) and adenovirus (14.3%). A total of 481 cases (29.82%) were MP-positive. The co-infection rate was 32.18% (519 cases), and the mixed infections of bacteria and viruses were common (47.4%).

CONCLUSIONSRSV and Hemophilus parainfluenzae are the major pathogens of CAP in children from the Chongqing area. MP is also an important pathogen. The co-infection of bacteria and viruses is prevalent.

Adolescent ; Child ; Child, Preschool ; Community-Acquired Infections ; etiology ; Female ; Haemophilus parainfluenzae ; isolation & purification ; Hospitalization ; Humans ; Infant ; Male ; Mycoplasma pneumoniae ; isolation & purification ; Pneumonia ; etiology ; Respiratory Syncytial Viruses ; isolation & purification

Toothbrushes play an essential role in oral hygiene. However, they can be significant in microbial transmission and can increase the risk of infection, since they can serve as a reservoir for microorganisms in healthy, oral-diseased and medically ill adults. This study was conducted to evaluate toothbrush contamination in six toothbrushes donated from four people. Two participants each supplied two toothbrushes - one used in the bathroom and one used in the workplace. The other two people each donated two toothbrushes used in the workplace. Polymerase chain reaction was used to construct a 16S rRNA clone library. Sequences of cloned DNA were compared with those from the reference organisms provided by GenBank. A total 120 clones, representing 20 clones for each toothbrush, were analyzed. They are composed of six pylum, 46 genera and 79 species. The most dominant species were Streptococcus oralis, Streptococcus parasanguinis and Haemophilus parainfluenzae. Enterobacter and Escherichia were recovered from toothbrushes used domestically. Toothbrushes used in the workplace did not contain Enterobacteria.
Adult ; Bacteria ; Clone Cells ; Databases, Nucleic Acid ; DNA ; Enterobacter ; Enterobacteriaceae ; Escherichia ; Haemophilus parainfluenzae ; Humans ; Oral Hygiene ; Polymerase Chain Reaction ; Streptococcus ; Streptococcus oralis

Prosthetic valve endocarditis (PVE) caused by Haemophilus parainfluenzae (H. parainfluenzae) is very rare. Here, we report a case of H. parainfluenzae PVE that developed following the Bentall procedure complicated by a pseudoaneurysm and cerebral emboli. A diagnosis was delayed in this case because of the slow-growing nature of the organism and the unusual clinical presentation.
Aneurysm, False* ; Diagnosis ; Endocarditis* ; Haemophilus parainfluenzae* ; Heart Valve Prosthesis ; Intracranial Embolism ; Paramyxoviridae Infections

Prosthetic valve endocarditis (PVE) caused by Haemophilus parainfluenzae (H. parainfluenzae) is very rare. Here, we report a case of H. parainfluenzae PVE that developed following the Bentall procedure complicated by a pseudoaneurysm and cerebral emboli. A diagnosis was delayed in this case because of the slow-growing nature of the organism and the unusual clinical presentation.
Aneurysm, False* ; Diagnosis ; Endocarditis* ; Haemophilus parainfluenzae* ; Heart Valve Prosthesis ; Intracranial Embolism ; Paramyxoviridae Infections

OBJECTIVETo establish multiplex PCR-based assays for detecting H.influenzae and H.parainfluenzae. And the PCR-based assays were applied to detect the carriage rates of H.influenzae and H.parainfluenzae in nasopharyngeal swab specimens which were collected from healthy children.

METHODSMultiplex primers for species-specific PCR were designed by using DNAstar soft based on the sequences of 16S rRNA genes from genus Haemophilus to detect H.influenzae and H.parainfluenzae.

RESULTSThe sensitivity of the 16S rRNA PCR assay for detecting H.influenzae and H.parainfluenzae was 97.53% and 100% respectively, and the specificity was 95.89% and 96.63% respectively. Youden's Index on the ability to detect H.influenzae and H.parainfluenzae was 0.9342 and 0.9663 respectively. 666 nasopharyngeal swab specimens were collected from healthy children. The detection rates of H.influenzae and H.parainfluenzae were 14.11% and 16.07% respectively by using isolation and culture methods. The detection rates of H.influenzae and H.parainfluenzae were 43.54% and 57.96% respectively by 16S rRNA PCR assays. The carriage rates of serotypes a, b, c, d, e, f and non-typeable isolates were 0% (0/666), 0.15% (1/666), 1.20% (8/666), 0.15% (1/666), 1.20% (8/666), 1.80% (12/666), 95.50% (636/666) respectively.

CONCLUSIONThe multiplex PCR assays were very rapid, reliable and feasible methods for detection of H.influenzae and H.parainfluenzae in pharyngeal swab specimens which were compared to conventional isolation and culture methods. 95.5% of H.influenzae strains in healthy children were nontypeable. The encapsulated or typable strains were mainly three serotypes which was c, e, and f serotype.

Haemophilus influenzae ; classification ; genetics ; isolation & purification ; Haemophilus parainfluenzae ; classification ; genetics ; isolation & purification ; Humans ; Multiplex Polymerase Chain Reaction ; methods ; Nasopharynx ; microbiology ; RNA, Bacterial ; genetics ; RNA, Ribosomal, 16S ; genetics ; Sensitivity and Specificity

The HACEK group of microorganisms is responsible for approximately 3-6% of endocarditis cases and is a major cause of culture-negative endocarditis. Here, we report a case of Haemophilus parainfluenzae infective endocarditis that was diagnosed by direct PCR sequencing of 16S rRNA from resected vegetation. A healthy 26-yr-old man was admitted to the emergency room (ER) on March 27, 2011 because of intermittent high fever. The patient was prescribed cefpodoxime for 5 days at the ER. Six and 11 sets of blood cultures were performed at the ER and in a general ward, respectively, using BACTEC Plus Aerobic/F (Becton-Dickinson, USA) and Lytic Anaerobic/F Plus (BD) together. Echocardiography revealed a large vegetation at the posterior mitral valve leaflet. After performing mitral valvoplasty on hospital day (HD) 11, the vegetation tissue was cultured in thioglycolate broth, blood agar, Brucella agar, and MacConkey agar for 7 days, but no organism was grown. Direct PCR sequencing of 16S rRNA of the tissue revealed the presence of H. parainfluenzae. In the 17 sets of blood cultures, bacterial growth was detected in only 2 aerobic bottles of 5 sets taken at HD 9 after 10-day and 14-day incubation. The organism was identified as H. parainfluenzae by using the VITEK NHI card (bioMerieux, France). Direct PCR sequencing of vegetation could be useful in diagnosing bacterial pathogens in infective endocarditis patients, especially in culture-negative cases.
Agar ; Brucella ; Ceftizoxime ; Echocardiography ; Emergencies ; Endocarditis ; Fever ; Haemophilus ; Haemophilus parainfluenzae ; Humans ; Mitral Valve ; Paramyxoviridae Infections ; Patients' Rooms ; Polymerase Chain Reaction ; Sequence Analysis, RNA

Blood culture-negative infective endocarditis (CNE) can be a diagnostic dilemma. Herein, we report a case of CNE caused by Haemophilus parainfluenzae identified only via 16S rRNA sequence analysis directly from valve tissue. A 17-year-old boy presented with high spiking fever for one month. Pansystolic murmur (Grade III) and vegetation (0.65x0.26 cm and 0.62x0.55 cm) on the anterior mitral valve leaflet via transesophageal echocardiogram suggested the diagnosis of infective endocarditis (IE). However, blood culture performed on admission was negative even after 2 weeks of incubation. Gram stain and culture of a direct tissue specimen failed to identify causative microorganism, while 16S rRNA gene sequences (548 bp) showed 100% identity with those of Haemophilus parainfluenzae (GenBank: FJ939586.1). The 16S rRNA sequence analysis with a direct tissue specimen might be useful in cases of CNE.
Endocarditis ; Fever ; Genes, rRNA ; Haemophilus ; Haemophilus parainfluenzae ; Mitral Valve ; Sequence Analysis

Haemophilus parainfluenzae, one of the member of the HACEK group of gram-negative oropharyngeal species, is a rare cause of subacute native valve endocarditis. Infective endocarditis caused by H. parainfluenzae appears to carry a high incidence rate of cerebral embolism, often making the timing of surgical intervention difficult. A 52-year-old male was diagnosed with acute endocarditis caused by H. parainfluenzae complicated with acute cerebral infarctions. After institution of antibiotic therapy, this patient was mechanically ventilated because of the sudden onset of dyspnea. Repeated two-dimensional echocardiography demonstrated rate-dependent mitral stenosis without interval change of vegetations on the mitral valve. Making a decision regarding the timing of surgical intervention was difficult because of a fear of clinical deterioration after early valve replacement.
Cerebral Infarction ; Dyspnea ; Echocardiography ; Endocarditis ; Endocarditis, Bacterial ; Haemophilus ; Haemophilus parainfluenzae ; Heart Valve Prosthesis Implantation ; Humans ; Incidence ; Intracranial Embolism ; Male ; Middle Aged ; Mitral Valve ; Mitral Valve Stenosis ; Paramyxoviridae Infections