Identified micro-organisms in hospitalized community-acquired pneumonia patients living near goat and poultry farms

Background In the Netherlands, an increased risk of community-acquired pneumonia (CAP) has been reported for adults living near goat and poultry farms. Previous results of respiratory microbiome studies in hospitalized CAP patients near poultry farms suggested a higher relative abundance of Streptococcus pneumoniae. This retrospective study, using routine laboratory data from hospitalized CAP patients, aims to explore possible aetiologic micro-organisms of CAP in relation to livestock exposure. Methods Patient characteristics and PCR and urinary antigen test results were retrieved retrospectively from electronic medical records of CAP patients admitted to the Jeroen Bosch Hospital or Gelre Hospital in the Netherlands during 2016–2017. Distances between the patients’ home address and the nearest poultry and goat farm were calculated. Differences in laboratory test results between CAP patients with and without goat or poultry farms within 2 km of their home address were analyzed using Fisher’s exact test. Results In total, 2230 CAP episodes with diagnostic results were included. In only 25% of the CAP episodes, a micro-organism was detected. A positive urinary antigen test for S. pneumoniae was found more often in patients living within two kilometers of goat (15.2% vs. 11.3%) and poultry farms (14.4% vs. 11.3%), however these differences were not statistically significant (p = 0.1047 and p = 0.1376). Conclusion Our retrospective analysis did not show statistically significant differences in the identified micro-organisms in hospitalized CAP patients related to livestock farming. The study was hampered by limited statistical power and limited laboratory results. Therefore, the potential increased CAP risk around goat and poultry farms will be further explored in a prospective study among CAP patients in primary care.

A community-acquired Legionnaires’ disease outbreak caused by Legionella pneumophila serogroup 2: an uncommon event, Italy, August to October 2018

In September 2018 in Brescia province, northern Italy, an outbreak of Legionnaires' disease (LD) caused by Legionella pneumophila serogroup 2 (Lp2) occurred. The 33 cases (two fatal) resided in seven municipalities along the Chiese river. All cases were negative by urinary antigen test (UAT) and most were diagnosed by real-time PCR and serology. In only three cases, respiratory sample cultures were positive, and Lp2 was identified and typed as sequence type (ST)1455. In another three cases, nested sequence-based typing was directly applied to respiratory samples, which provided allelic profiles highly similar to ST1455. An environmental investigation was undertaken immediately and water samples were collected from private homes, municipal water systems, cooling towers and the river. Overall, 533 environmental water samples were analysed and 34 were positive for Lp. Of these, only three samples, all collected from the Chiese river, were Lp2 ST1455. If and how the river water could have been aerosolised causing the LD cases remains unexplained. This outbreak, the first to our knowledge caused by Lp2, highlights the limits of UAT for LD diagnosis, underlining the importance of adopting multiple tests to ensure that serogroups other than serogroup 1, as well as other Legionella species, are identified.

Detection of Streptococcus pneumoniae in Urine by Loop-Mediated Isothermal Amplification

Objective To assess the loop-mediated isothermal amplification (LAMP) to detect cell-free DNA from Streptococcus pneumoniae in urine samples from children with pneumococcal pneumonia. Methods LAMP reactions using four primers (backward inner primer, forward inner primer, B3, and F3) targeting conserved regions of the S. pneumoniae ply gene and DNA from the recombinant plasmid pTrc99A-ply were optimized for temperature (65°C) and MgSO4 concentration (8 mM) conditions. Urine samples from 71 patients with symptoms of pneumonia and from 17 healthy children were tested side by side using the isothermal methodology LAMP and the commercial urinary antigen test, BinaxNOW S. pneumoniae assay. Percentages of sensitivity, specificity, positive predictive value (PPV), negative predictive value, and positive (LR) were calculated to compare both tests. Results The specificity of the LAMP reaction was confirmed against several species of bacteria and yeast that can cause pneumonia or urine infections. The suitability of the LAMP assay was evaluated in urine samples from 71 patients and 17 healthy children. All patients (100%) with confirmed pneumococcal pneumonia were positive for the LAMP assay. Among patients with possible/probable pneumonia, 74.1% were identified as positive using the LAMP test. Notably, a higher specificity (95.4%), PPV (94.1%) and positive LR (21.7) were found compared with the urinary antigen test. Conclusion The presence of S. pneumoniae cell-free DNA in urine samples of pediatric patients can be used as a specific diagnostic biomarker for community-acquired pneumonia by using the LAMP methodology.