Legionella pneumophila is an environmental bacterium, an opportunistic premise plumbing pathogen that causes the Legionnaires’ disease. L. pneumophila presents a serious health hazard in building water systems, due to its high resistance to standard water disinfection methods. Our aim was to study the use of photodynamic inactivation (PDI) against Legionella. We investigated and compared the photobactericidal potential of five cationic dyes. We tested toluidine blue (TBO) and methylene blue (MB), and three 3-N-methylpyridylporphyrins, one tetra-cationic and two tri-cationic, one with a short (CH3) and the other with a long (C17H35) alkyl chain, against L. pneumophila in tap water and after irradiation with violet light. All tested dyes demonstrated a certain dark toxicity against L. pneumophila; porphyrins with lower minimal effective concentration (MEC) values than TBO and MB. Nanomolar MEC values, significantly lower than with TBO and MB, were obtained with all three porphyrins in PDI experiments, with amphiphilic porphyrin demonstrating the highest PDI activity. All tested dyes showed increasing PDI with longer irradiation (0–108 J/cm2), especially the two hydrophilic porphyrins. All three porphyrins caused significant changes in cell membrane permeability after irradiation and L. pneumophila, co-cultivated with Acanthamoeba castellanii after treatment with all three porphyrins and irradiation, did not recover in amoeba. We believe our results indicate the considerable potential of cationic porphyrins as effective anti-Legionella agents.

Considering the fact that water is a basic need of every living being, it is important to ensure its safety. In this work, the data on the presence of the opportunistic pathogen P. aeruginosa in drinking water (n = 4171) as well as in pool water (n = 5059) in Primorje-Gorski Kotar County in Croatia in the five-year period (2016–2020) were analysed. In addition, the national criteria were compared with those of neighboring countries and worldwide. The proportion of P. aeruginosa-positive samples was similar for drinking water (3.9%) and pool water (4.6%). The prevalence of this bacterium was most pronounced in the warmer season. P. aeruginosa-positive drinking water samples were mostly collected during building commissioning, while pool samples were from entertainment and spa/hydromassage pools. Outdoor pools showed a higher percentage of positive samples than indoor pools, as well as the pools filled with freshwater rather than seawater. The highest P. aeruginosa load was found in rehabilitation pools. Croatia, Serbia and Montenegro are countries that have included P. aeruginosa in their national regulations as an indicator of the safety of water for human consumption as well as for bottled water, while Slovenia and Bosnia and Herzegovina have limited this requirement to bottled water only. In the case of swimming pool water, this parameter is mandatory in all countries considered in this study.

Bacterial pneumonia caused by the inhalation of aerosols contaminated with Legionella spp. is also known as Legionnaires' disease. In this study, we report a case of pneumonia caused by Legionella pneumophila sg.1 in a 58-year-old man who visited a sea water-filled whirlpool within a hotel and spa complex. The patient's Legionella urine antigen test was positive for L. pneumophila sg.1. During the field study, samples were taken from both the outdoor and indoor sea water-filled pools. Samples from the whirlpool were culture positive for L. pneumophila sg.1. Typing results indicated sea water isolate belonged to Sequence type ST82 and Allentown/France MAb subgroup. In vitro experiments showed that L. pneumophila strains are able to survive within sea water up to 7 days, and survival time is prolonged with sea water dilution. Also, our results indicate that L. pneumophila Allentown strain was the most resistant to adverse conditions in sea water with the highest values of DT50 (420 min) and DT90 (1,396 min). The possible source of infection was adding potable water for filling up the whirlpool. The survival of the L. pneumophila in additionally conditioned sea water should be considered in a further study.

Cadmium occurs naturally in the environment and as an anthropogenic pollutant. Exposure to low concentrations of cadmium is inevitable and may produce toxic effects. Another important aspect of cadmium toxicity is its interaction, often antagonistic, with essential elements such as selenium. The aim of this study was to highlight the risks of long-term exposure to low cadmium concentrations, using a scientific and chemical approach and hares (Lepus europaeus Pallas) as model organisms in a field study. Two study areas were monitored. Levels of cadmium and selenium were quantified in the organs of hares, the expression of metallothioneins I + II and the products of lipid peroxidation were determined. The median cadmium concentrations (wet weight) in the muscle, liver, kidney and brain of hares from an exposed group ranged from 0.033 to 0.037, 0.763 to 1.054, 3.090 to 16.594 and 0.016 to 0.087 µg g(-1), respectively; whereas, the median selenium concentrations (wet weight) ranged from 0.100 to 0.108, 0.153 to 0.332, 0.677 to 0.701 and 0.078 to 0.116 µg g(-1), respectively. Expression of the metallothioneins I + II proteins was observed in tissues. Lipid peroxidation (LPO) levels, measured as malondialdehyde (MDA) equivalents, increased with the cadmium concentration. Further research on long-term exposure to low concentrations of cadmium in the environment is needed.

The aim of this study was to analyze and evaluate risks of long-term exposure to mercury in hares (Lepus europaeus Pallas), with a chemical-analytical approach evaluating median mass fraction of toxic mercury in the hares organs (liver, kidney, muscle and brain). To obtain better insight into possible effects of mercury, the study included screening of the oxidative status after long term exposure to low concentrations of mercury. Hares organs were analyzed for total mercury concentration by AAS. Glutathione and glutathione-dependent enzymes status was also investigated. The median mercury concentrations (wet weight) in the liver, kidney, muscle and brain of the hares ranged from 0.058-0.189, 0.138-0.406, 0.013-0.046 and 0.022-0.102 μg/g respectively. Concentration of the glutathione (GSH), glutathione-peroxidase (GPx), and glutathione-reductase (GR) activity increased with the mercury concentration. However, glutathione S-transferase (GST) and superoxide-dismutase (SOD) activity decreased with the mercury concentration. The results of this study show the impact of environmentally absorbed mercury on the antioxidant status of the examined hares. Further research on long-term exposure to low concentrations of mercury is needed.