TY - JOUR

T1 - Lessons learned from a one-year study of Legionella spp. cultivation from activated sludge samples

AU - Dörrie, Beatriz

AU - Nogueira, Regina

N1 - Publisher Copyright: © 2024 The Authors

PY - 2024/10

Y1 - 2024/10

N2 - Risk assessment and management of Legionella spp. contamination in activated sludge in wastewater treatment plants is carried out using the culture method. Underestimation of Legionella spp. is frequently reported in the literature, but a comprehensive long-term study of the performance of the method under comparable conditions is still lacking. The aim of this study is to evaluate the recovery rate and limit of detection of the culture method for Legionella spp. from activated sludge samples collected during the different seasons of the year. Activated sludge samples spiked with Legionella pneumophila subsp. pneumophila strain Philadelphia-1 (mean concentration 5.2 ± 0.35 logCFU/mL) were analysed monthly for one year using the culture method. Three different sample pre-treatments were compared, namely filtration, acid treatment and thermal treatment, and the recovery rate and limit of detection were assessed for each. The recovery rate of the culture method for Legionella spp. depended on the type of sample pre-treatment and the season of activated sludge sampling, while the limit of detection depended only on the sample pre-treatment. The best performance of the culture method, defined as the combination of the highest recovery rate and lowest limit of detection, was obtained for the filtered acid pre-treated samples (recovery rate: 89 ± 4 %; limit of detection: 1.3 logCFU/mL in 83 % of the samples). The lowest limit of detection was observed for the filtered thermally pre-treated samples (1.0 logCFU/mL in 93 % of the samples). Simultaneously, both thermally pre-treated samples showed up to a third lower recovery rates than the other pre-treatments in winter, while untreated and acid pre-treated samples showed consistently high recovery rates (>80%, logCFU/mL). The recovery rates of the unfiltered and filtered thermally pre-treated samples showed significant weak to strong positive correlations with the organic and phosphorus load in the influent as well as with the water and atmospheric temperatures, indicating that the recovery rate depends on the seasonal variation of the wastewater composition. This study presents new insights into the detection and quantification of Legionella spp. in activated sludge samples and considers seasonal dependencies in analytical results.

AB - Risk assessment and management of Legionella spp. contamination in activated sludge in wastewater treatment plants is carried out using the culture method. Underestimation of Legionella spp. is frequently reported in the literature, but a comprehensive long-term study of the performance of the method under comparable conditions is still lacking. The aim of this study is to evaluate the recovery rate and limit of detection of the culture method for Legionella spp. from activated sludge samples collected during the different seasons of the year. Activated sludge samples spiked with Legionella pneumophila subsp. pneumophila strain Philadelphia-1 (mean concentration 5.2 ± 0.35 logCFU/mL) were analysed monthly for one year using the culture method. Three different sample pre-treatments were compared, namely filtration, acid treatment and thermal treatment, and the recovery rate and limit of detection were assessed for each. The recovery rate of the culture method for Legionella spp. depended on the type of sample pre-treatment and the season of activated sludge sampling, while the limit of detection depended only on the sample pre-treatment. The best performance of the culture method, defined as the combination of the highest recovery rate and lowest limit of detection, was obtained for the filtered acid pre-treated samples (recovery rate: 89 ± 4 %; limit of detection: 1.3 logCFU/mL in 83 % of the samples). The lowest limit of detection was observed for the filtered thermally pre-treated samples (1.0 logCFU/mL in 93 % of the samples). Simultaneously, both thermally pre-treated samples showed up to a third lower recovery rates than the other pre-treatments in winter, while untreated and acid pre-treated samples showed consistently high recovery rates (>80%, logCFU/mL). The recovery rates of the unfiltered and filtered thermally pre-treated samples showed significant weak to strong positive correlations with the organic and phosphorus load in the influent as well as with the water and atmospheric temperatures, indicating that the recovery rate depends on the seasonal variation of the wastewater composition. This study presents new insights into the detection and quantification of Legionella spp. in activated sludge samples and considers seasonal dependencies in analytical results.

KW - Acid and thermal pre-treatment

KW - Activated sludge

KW - Culture method

KW - Legionella pneumophila

KW - Legionella spp

KW - Wastewater treatment plant

UR - http://www.scopus.com/inward/record.url?scp=85202347069&partnerID=8YFLogxK

U2 - 10.1016/j.jenvman.2024.122266

DO - 10.1016/j.jenvman.2024.122266

M3 - Article

AN - SCOPUS:85202347069

JO - Journal of Environmental Management

JF - Journal of Environmental Management

SN - 0301-4797

IS - 369

M1 - 122266

ER -