Objective: To analyze the pollution status and influencing factors of Legionella pneumophila in a secondary water supply facility in a city. Methods: From June to August 2020, a survey on the level of Legionella pneumophila in secondary water supply unit was carried out in a city in northern China, and 304 sets of secondary water supply facilities were included in the study. A total of 760 water samples were collected from the inlet and outlet water of the secondary water supply facilities and some water samples in the water tank were collected for the detection of Legionella pneumophila, standard plate-count bacteria and related physical and chemical indicators. Through questionnaire survey, the basic information of secondary water supply facilities and daily management of water quality were collected. Multivariate logistic regression model was used to analyze the influencing factors of Legionella pneumophila contamination. Results: Among 304 sets of secondary water supply facilities, most of them were located in residential buildings [57.24% (174/304)]. High and low water tank water supply, low water tank variable frequency conversion water supply and non-negative pressure water supply accounted for 26.6% (81/304), 36.8% (112/304) and 36.5% (111/304), respectively. About 25.7% of facilities (78/304) were positive for Legionella pneumophila. Among them, the positive rates of Legionella pneumophila in high and low water tank water supply, low water tank variable frequency conversion water supply and non-negative pressure water supply facilities were 38.3% (31/81), 29.5% (33/112) and 12.6% (14/111), respectively. The results of multivariate logistic regression model analysis showed that the disinfectant residue could reduce the risk of Legionella pneumophila contamination in water samples, and the OR (95%CI) value was 0.083 (0.022-0.317). The increase of the standard plate-count bacteria and conductivity might increase the risk of Legionella pneumophila contamination in water samples. The OR (95%CI) values were 3.160 (1.667-5.99) and 1.004 (1.001-1.006), respectively. Compared with the non-negative pressure water supply, the risk of Legionella pneumophila contamination of secondary water supply facilities was increased by water supply from high and low water tanks and variable frequency conversion water supply from low water tanks, with OR (95%CI) values of 4.296 (2.096-8.803) and 2.894 (1.449-5.782), respectively. Conclusion: The positive rate of Legionella pneumophila in secondary water supply in the study city is high. Disinfectant residue, conductivity and method of water supply are associated with the positive rate of Legionella pneumophila.
Humans ; Legionella ; Water Microbiology ; Water Supply ; Legionella pneumophila ; Reproduction ; Disinfectants

OBJECTIVE: To investigate the molecular characteristics and intracellular growth ability of Legionella pneumophila (L. pneumophila) strains from 1989 to 2016 in Sichuan Province, China. METHODS: Seventy-nine isolates of L. pneumophila were collected from environmental and clinical sources, including cooling towers, hot springs, bath water, fountains, and patients, and identified with 16S rRNA gene analysis and serum agglutination assay. The isolates were then typed by Sequence-Based Typing (SBT), and Genotyping of forty-two LP1 strains were analyzed by means of multiple-locus VNTR analysis with 8 loci (MLVA-8). All strains were further analyzed for two virulence genes: Legionella vir homologue (lvh) and repeats in structural toxin (rtxA). The intracellular growth ability of 33 selected isolates was determined by examining their interaction with J774 cells. RESULTS: All isolates were identified to L. pneumophila including 11 serogroups, among which the main serogroup were LP1, accounting for 54.43%. Thirty-three different sequence types (STs) from five main clonal groups and five singletons were identified, along with 8 different MLVA patterns. Both the lvh and rtxA loci were found in all 79 strains. Thirty isolates showed high intracellular growth ability in J774 cells. CONCLUSION L. pneumophila is a potential threat to public health, and effective control and prevention strategies are urgently needed.
Bacterial Proteins ; genetics ; Bacterial Toxins ; genetics ; China ; Genotyping Techniques ; Humans ; Legionella pneumophila ; genetics ; growth & development ; isolation & purification ; RNA, Ribosomal, 16S ; genetics ; Water Microbiology

BACKGROUND/AIMS: To use serological and multiplex polymerase chain reaction (PCR) assays to examine sputum samples from patients experiencing acute exacerbation of chronic obstructive pulmonary disease (AECOPD) for the presence of atypical pathogens, including Mycoplasma pneumoniae, Chlamydia pneumoniae, and Legionella pneumophila. METHODS: From September 2012 to February 2014, 341 patients with AECOPD attending outpatient clinics were enrolled as part of a randomized, double-blind, multicenter study. A commercial enzyme-linked immunosorbent assay was used to measure serum immunoglobulin M (IgM) and IgG antibody titers on the first day of the study and at 36 days post-enrollment. Multiplex PCR was used to test sputum samples for the presence of atypical pathogens. A urinary antigen test for L. pneumophila was performed on the first day. RESULTS: Nineteen patients (5.6%) showed serological evidence of acute infection with M. pneumoniae. Also, one and seven patients (2%) showed serological evidence of acute infection with C. pneumoniae and L. pneumophila, respectively. All DNA samples were negative for M. pneumoniae, C. pneumoniae, and L. pneumophila according to PCR. Only one urine sample was positive for L. pneumophila antigen, but serologic evidence was lacking. CONCLUSIONS: Serological testing suggested that infection by atypical pathogens during AECOPD was relatively uncommon. In addition, PCR provided no direct evidence of infection by atypical pathogens. Thus, atypical pathogens may not be a major cause of AECOPD in South Korea.
Ambulatory Care Facilities ; Chlamydophila pneumoniae ; DNA ; Enzyme-Linked Immunosorbent Assay ; Humans ; Immunoglobulin G ; Immunoglobulin M ; Korea ; Legionella pneumophila ; Multiplex Polymerase Chain Reaction ; Mycoplasma pneumoniae ; Pneumonia ; Pneumonia, Mycoplasma ; Polymerase Chain Reaction* ; Pulmonary Disease, Chronic Obstructive* ; Serologic Tests ; Sputum

BACKGROUND: Infection by the intracellular bacteria Mycoplasma pneumoniae, Chamydophila pneumoniae, and Legionella pneumophila are common causes of community-acquired pneumonia (CAP). This study describes the evaluation of a new multiplex real-time PCR test, EuDx™-PN MLC Detection Kit (EUDIPIA), which allows the simultaneous detection of M. pneumoniae, C. pneumoniae, and L. pneumophila in respiratory samples. METHODS: A total of 353 samples were tested using three PCR kits: multiplex PCR (Seeplex PneumoBacter ACE Detection Kit) and two multiplex real-time PCR (EuDx™-PN MLC Detection Kit and Anyplex™ II RB5 Detection Kit). The results were considered true positives (expanded standard) for M. pneumoniae, C. pneumoniae, and L. pneumophila if they were positive according to any of the three tests. RESULTS: The sensitivity and specificity of EuDx™-PN MLC Detection Kit were 93.3–100% and 100%, respectively. The agreement rate and Cohen's kappa coefficient (value) between EuDx™-PN MLC Detection Kit and Anyplex™ II RB5 Detection Kit for M. pneumoniae, C. pneumoniae, and L. pneumophila were 70–100% and 0.82–1, respectively. CONCLUSION: These results demonstrate that the EuDx™-PN MLC Detection Kit is a sensitive, specific, and useful screening tool for the detection of atypical pathogens in respiratory samples and can be helpful in selecting appropriate antimicrobial therapy for patients with respiratory infection.
Bacteria ; Chlamydial Pneumonia* ; Chlamydophila pneumoniae* ; Chlamydophila* ; Humans ; Legionella pneumophila* ; Legionella* ; Mass Screening ; Multiplex Polymerase Chain Reaction ; Mycoplasma pneumoniae* ; Mycoplasma* ; Pneumonia ; Pneumonia, Mycoplasma* ; Polymerase Chain Reaction ; Real-Time Polymerase Chain Reaction ; Respiratory Tract Infections ; Sensitivity and Specificity

OBJECTIVES: Legionnaires' disease (LD) is a severe type of pneumonia caused by inhalation of aerosols contaminated with Legionella. On September 22, 2016, a single case of LD was reported from a newly built apartment building in Gyeonggi province. This article describes an epidemiologic investigation of LD and identification of the possible source of infection. METHODS: To identify the source of LD, we interviewed the patient's husband using a questionnaire based on the Legionella management guidelines from the Korea Centers for Disease Control and Prevention. Water samples from the site were collected and analyzed. An epidemiological investigation of the residents and visitors in the apartment building was conducted for 14 days before the index patient's symptoms first appeared to 14 days after the implementation of environmental control measures. RESULTS: Legionella pneumophila serogroup 1 was isolated from the heated-water samples from the patient's residence and the basement of the apartment complex. Thirty-two suspected cases were reported from the apartment building during the surveillance period, yet all were confirmed negative based on urinary antigen tests. CONCLUSIONS: The likely source of infection was the building's potable water, particularly heated water. Further study of effective monitoring systems in heated potable water should be considered.
Aerosols ; Centers for Disease Control and Prevention (U.S.) ; Disease Outbreaks ; Drinking Water ; Gyeonggi-do ; Hot Temperature ; Humans ; Inhalation ; Korea ; Legionella ; Legionella pneumophila ; Legionnaires' Disease ; Pneumonia ; Serogroup ; Spouses ; Water

OBJECTIVETo investigate the distribution characteristics of Mycoplasma pneumoniae (MP), Chlamydia pneumoniae (CP), and Legionella pneumophila (LP) in hospitalized children with acute respiratory tract infection (ARTI).

METHODSA total of 13 198 hospitalized children with ARTI were enrolled as study subjects. Whole blood and urine were collected. The passive agglutination was used to detect serum MP-IgM, ELISA was used to detect serum CP-IgM, and immunochromatography was performed to detect urinary LP antigen.

RESULTSAmong the 13 198 hospitalized ARTI children, the detection rates of MP, CP, and LP were 25.31%, 12.74% and 3.27%, suggesting that MP had the highest detection rate (P<0.0125). The detection rates of MP in 2013 and 2014 were significantly higher than that in 2012 (P<0.0125). CP had the highest detection rate in 2013, and LP had the highest detection rate in 2014 (P<0.0125). These three pathogens were detected all around the year, and MP had the highest detection rate in all seasons (P<0.0125). The detection rate of mixed infection with three pathogens was 4.35%, and mixed infection with MP and CP was the most common (P<0.0071). Among the children in different age groups, the patients aged 5-16 years showed the highest overall detection rate of three pathogens (P<0.0071). Among the children with different types of ARTI, the children with bronchopneumonia showed the highest overall detection rate of three pathogens (P<0.0045).

CONCLUSIONSMP, CP, and LP, particularly MP, are important pathogens for children with ARTI in the local area. LP infection tends to increase year by year and should be taken seriously in clinical practice.

Acute Disease ; Adolescent ; Child ; Child, Hospitalized ; Child, Preschool ; Chlamydophila pneumoniae ; isolation & purification ; Female ; Humans ; Infant ; Infant, Newborn ; Legionella pneumophila ; isolation & purification ; Mycoplasma pneumoniae ; isolation & purification ; Pregnancy ; Respiratory Tract Infections ; microbiology

The role of atypical bacteria and the effect of antibiotic treatments in acute bronchitis are still not clear. This study was conducted at 22 hospitals (17 primary care clinics and 5 university hospitals) in Korea. Outpatients (aged > or = 18 yr) who had an acute illness with a new cough and sputum (< or = 30 days) were enrolled in 2013. Multiplex real-time polymerase chain reaction (RT-PCR) was used to detect five atypical bacteria. A total of 435 patients were diagnosed as having acute bronchitis (vs. probable pneumonia, n = 75), and 1.8% (n = 8) were positive for atypical pathogens (Bordetella pertussis, n = 3; B. parapertussis, n = 0; Mycoplasma pneumoniae, n = 1; Chlamydophila pneumoniae, n = 3; Legionella pneumophila, n = 1). Among clinical symptoms and signs, only post-tussive vomiting was more frequent in patients with atypical pathogens than those without (P = 0.024). In all, 72.2% of the enrolled patients received antibiotic treatment at their first visits, and beta-lactams (29.4%) and quinolones (20.5%) were the most commonly prescribed agents. In conclusion, our study demonstrates that the incidence of atypical pathogens is low in patients with acute bronchitis, and the rate of antibiotic prescriptions is high.
Anti-Bacterial Agents/therapeutic use ; Bordetella parapertussis/genetics/*isolation & purification ; Bordetella pertussis/genetics/*isolation & purification ; Bronchitis/drug therapy/*microbiology ; Chlamydophila pneumoniae/genetics/*isolation & purification ; Community-Acquired Infections/microbiology ; Female ; Humans ; Hypertension/complications ; Legionella pneumophila/genetics/*isolation & purification ; Male ; Middle Aged ; Mycoplasma pneumoniae/genetics/*isolation & purification ; Real-Time Polymerase Chain Reaction ; Republic of Korea ; Sputum/microbiology

The harshness of legionellosis differs from mild Pontiac fever to potentially fatal Legionnaire's disease. The increasing development of drug resistance against legionellosis has led to explore new novel drug targets. It has been found that phosphoglucosamine mutase, phosphomannomutase, and phosphoglyceromutase enzymes can be used as the most probable therapeutic drug targets through extensive data mining. Phosphoglucosamine mutase is involved in amino sugar and nucleotide sugar metabolism. The purpose of this study was to predict the potential target of that specific drug. For this, the 3D structure of phosphoglucosamine mutase of Legionella pneumophila (strain Paris) was determined by means of homology modeling through Phyre2 and refined by ModRefiner. Then, the designed model was evaluated with a structure validation program, for instance, PROCHECK, ERRAT, Verify3D, and QMEAN, for further structural analysis. Secondary structural features were determined through self-optimized prediction method with alignment (SOPMA) and interacting networks by STRING. Consequently, we performed molecular docking studies. The analytical result of PROCHECK showed that 95.0% of the residues are in the most favored region, 4.50% are in the additional allowed region and 0.50% are in the generously allowed region of the Ramachandran plot. Verify3D graph value indicates a score of 0.71 and 89.791, 1.11 for ERRAT and QMEAN respectively. Arg419, Thr414, Ser412, and Thr9 were found to dock the substrate for the most favorable binding of S-mercaptocysteine. However, these findings from this current study will pave the way for further extensive investigation of this enzyme in wet lab experiments and in that way assist drug design against legionellosis.
Computer Simulation* ; Data Mining ; Drug Delivery Systems ; Drug Design ; Drug Resistance ; Fever ; Legionella pneumophila* ; Legionellosis* ; Legionnaires' Disease ; Metabolism ; Phosphoglycerate Mutase

Endogenous lipoid pneumonia is an uncommon condition. This is a report of a 29-year-old woman diagnosed with endogenous lipoid pneumonia associated with Legionella pneumophila serogroup 1 infection. The patient's endogenous lipoid pneumonia resolved completely after treatment for Legionella pneumophila infection. This suggests that early diagnosis and aggressive treatment of the underlying infection may prevent any long-term sequelae of lipoid pneumonia.
Adult ; Anti-Bacterial Agents ; therapeutic use ; Aza Compounds ; therapeutic use ; Azithromycin ; therapeutic use ; Female ; Fluoroquinolones ; Humans ; Legionella pneumophila ; classification ; Legionnaires' Disease ; diagnosis ; drug therapy ; microbiology ; Pneumonia, Lipid ; diagnosis ; drug therapy ; microbiology ; Quinolines ; therapeutic use ; Treatment Outcome

Bacterial Proteins ; chemistry ; Bacterial Secretion Systems ; Crystallography, X-Ray ; Legionella pneumophila ; metabolism ; Models, Molecular ; Protein Structure, Tertiary