Free-Living Protozoa and Legionella spp. Coexistence and Bacterial Diversity in Drinking Water Systems in Apartment Buildings and Hotels in Riga and Its Surroundings

Free-living protozoa (FLP) can create biofilms in water supply systems and can harbor bacteria, which potentially can be pathogenic, such as Legionella spp. Each year there are more cases of legionellosis in Latvia, so this problem is actual: in 2019 there were 42 cases, but in 2024-88 cases. In this study, the investigated question of the coexistence of FLP and Legionella spp. and bacterial diversity in the drinking water supply systems of Riga, Salaspils, and Jurmala multiapartment buildings and hotels situated in Riga and Jurmala, identify the main FLP genus, and study factors associated with FLP and Legionella spp. occurrence. With microscopy, microbiological, and molecular biology methods, FLP and, specifically, free-living amoeba (FLA) were detected and identified, and Legionella spp. bacteria were isolated. Three FLP genera were identified, including Acanthamoeba, Vahlkampfia, and Hartmanella (Vermamoeba). In hot water, more FLP and Legionella co-existence occurrences were detected. In 64.7% of FLP-positive samples, Hartmanella (Vermamoeba) spp. was detected. Various potentially pathogenic bacteria, such as Coxiella, Leptospira, and Mycobacterium, were detected in the water sample DNA sequences. The average hot water temperature in Riga was lower than 50 degrees C, which is not enough to minimize the risk of the Legionella bacteria proliferation. The Shannon's index values showed that bacterial diversity was higher in cold water samples, and the Pearson test showed that the correlation between building floor and Legionella quantity is positive. In this study, we also discovered that differences in bacterial diversity between water samples from two Daugava River banks' water sources are not significant, but the biggest exception was a much higher percentage of Chaetonotida (hairybellies) in the left river bank samples. Noticeably, there are more Legionella and FLP-positive samples from the kitchen than from the apartment shower. Each hotel building from this study has its own similar bacterial diversity in its water supply system.