High-resolution wastewater-based surveillance of three influenza seasons (2022–2025) reveals distinct seasonal patterns of viral activity in Munich, Germany.
In the Northern Hemisphere, annual waves of influenza disease with
varying degrees of spread and severity are observed each winter. With
wastewater-based surveillance (WBS), including both centralized (one
wastewater treatment plant, WWTP) and decentralized (three sewers)
sampling, we aimed to detect differences in influenza viral copy numbers
in wastewater over time, to investigate (sub)-community transmission
within a city. A total of 313 grab/spot and composite samples were
collected in Munich, Germany, during three consecutive influenza seasons
(2022/23, 2023/24, and 2024/25) and were analyzed for influenza A virus
(IAV) and influenza B virus (IBV) nucleic acids using digital droplet
PCR (ddPCR). IAV and IBV wastewater copy numbers and citywide reported
influenza cases showed strong correlations in both sampling approaches,
suggesting the decentralized approach to be a reliable indicator of
infection trends across the city. The three influenza seasons analyzed
differed significantly in terms of their seasonal distribution, for
example, exhibiting a strong co-circulation of IAV and IBV only in the
2024/25 season. Only with wastewater analysis, we reveal a reporting
delay of influenza A cases at the beginning of the 2023/24 season.
Higher influenza copy numbers were detected in sewer samples compared to
the WWTP influent, likely due to viral decay. The study underscores the
potential of influenza WBS to enable detection of seasonal onset early,
identify local transmission patterns, and reveal underreporting in
routine surveillance systems.