Wastewater-based epidemiology has shown to be a promising and innovative approach to measure a wide variety of illicit drugs that are consumed in the communities. In the same way as for illicit drugs, wastewater-based epidemiology is a promising approach to understand the prevalence of viruses in a community-level.

Global water scarcity and antimicrobial resistance (AMR) represent two escalating crises that urgently demand integrated and effective solutions. While wastewater reuse is increasingly promoted as a strategy to alleviate water scarcity, conventional treatment processes often fail to eliminate persistent contaminants and antibiotic-resistant microorganisms. Cold plasma (CP), a non-thermal advanced oxidation process, has demonstrated the strong potential to simultaneously inactivate pathogens and degrade micropollutants. CP generates a diverse mix of reactive oxygen and nitrogen species (ROS and RNS), as well as UV photons and charged particles, capable of breaking down complex contaminants and inducing irreversible damage to microbial cells. Laboratory studies have reported bacterial log reductions ranging from 1 to >8–9 log₁₀, with Gram-negative species such as E. coli and Pseudomonas aeruginosa showing higher susceptibility than Gram-positive bacteria. The inactivation of endospores and mixed-species biofilms has also been achieved under optimized CP conditions. Viral inactivation studies, including MS2 bacteriophage and norovirus surrogates, have demonstrated reductions >99.99%, with exposure times as short as 0.12 s. CP has further shown the capacity to degrade antibiotic residues such as ciprofloxacin and sulfamethoxazole by >90% and to reduce ARGs (e.g., bla, sul, and tet) in hospital wastewater. This perspective critically examines the mechanisms and current applications of CP in wastewater treatment, identifies the operational and scalability challenges, and outlines a research agenda for integrating CP into future water reuse frameworks targeting AMR mitigation and sustainable water management.

Background Sequencing of SARS-CoV-2 PCR-positive samples was introduced in Slovenia in January 2021. Our surveillance programme comprised three complementary schemes: (A) non-targeted sequencing of at least 10% of samples, (B) sequencing of samples positive after PCR screening for variants of concern (VOC) and (C) sequencing as per epidemiological indication. Aim We present the analysis of cumulative data of the non-targeted surveillance of SARS-CoV-2 and variant-dependent growth kinetics for the five most common variants in Slovenia for the first 9 months of 2021. Methods SARS-CoV-2 PCR-positive samples, from January to September 2021, were selected for sequencing according to the national surveillance plan. Growth kinetics studies were done on Vero E6 cells. Results Altogether 15,175 genomes were sequenced and 64 variants were detected, of which three successively prevailed. Variant B.1.258.17 was detected in ca 80% of samples in January and was replaced, within 9 weeks, by the Alpha variant. The number of cases decreased substantially during the summer of 2021. However, the introduction of the Delta variant caused a fourth wave and completely outcompeted other variants. Other VOC were only detected in small numbers. Infection of Vero E6 cells showed higher replication rates for the variants Alpha and Delta, compared with B.1.258.17, B.1.258, and B.1.1.70, which dominated in Slovenia before the introduction of the Alpha and Delta variants. Conclusion Information on SARS-CoV-2 variant diversity provided context to the epidemiological data of PCR-positive cases, contributed to control of the initial spread of known VOC and influenced epidemiological measures.

Since COVID-19, mpox was the first emerging pathogen to have spread globally in 2022. Wastewater-based surveillance (WBS) has proven to be an efficient early warning system for detecting potential resurgences. This report aims to provide insight into the development and implementation of WBS of mpox in Slovenia and to incorporate the surveillance results into the development of public health interventions. WBS of mpox was conducted during the period from 1 June 2023 to 30 September 2023 at the wastewater treatment plant (WWTP) Ljubljana and WWTP Koper. The selected detection method of the monkeypox virus (MPXV) in the wastewater sample was based on PCR analysis. The implemented laboratory method showed that the sample preparation and concentration method enables a stable procedure for MPXV detection in wastewater samples. The laboratory analysis of wastewater samples from the selected WWTPs did not detect the MPXV during the monitoring period. In the event of MPXV detection in a wastewater sample, targeted public health interventions would be implemented, focusing on increasing awareness among the groups of men who have sex with other men and searching for positive mpox cases. We recommend that the developed system be retained in the case of an emergency epidemiological situation.

COVID-19 is a persistent and ongoing global pandemic caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Non-anthropogenic factors, such as weather conditions and air quality are possible predictors of respiratory diseases, such as COVID-19. Weather conditions may also be a direct cause of biological interactions between SARS-CoV-2 and humans and vary widely between regions. The course of an epidemic is determined by several factors, including demographic and environmental parameters, many of which have an unknown correlation with COVID-19. The goal of this study is to access the influence of ground surface particulate matter and weather parameters on the dissemination of COVID-19 in Ljubljana, Slovenia. Data on average, minimum and maximum daily temperature, relative humidity; precipitation; sun duration in hours; and daily particle matter (PM10 and PM2.5) was obtained from the Slovenian Environmental Agency. The dataset on the number of daily COVID-19 cases in Ljubljana was obtained from the National Institute of Public Health. Data was divided into three groups: entire dataset; partial lifting of measures (from 1st of June to 30th of September); and less stringent containment measures (from 1st of July to 30th of September). Spearman rank correlation was used to investigate the association between new daily COVID-19 cases and weather data. The weather data and the daily new COVID -19 cases did not show any significant correlation during the entire period under investigation, except for PM 10 (0.30). With the beginning of the less stringent containment measures, five of the six weather parameters correlated significantly with the daily cases of COVID-19. Relative humidity showed the highest correlation coefficient (0.48). The correlations observed are highly dependent on the local policies that were in force during the period under study. The interaction between weather conditions and human behaviour may also be an important factor in understanding the relationship between weather and the spread of COVID -19. (www.actabiomedica.it)

With the reclassification of COVID-19 as an endemic disease and the relaxation of measures, Slovenia needed a complementary system for monitoring SARS-CoV-2 infections. This article provides an overview of the epidemiological situation of SARS-CoV-2 in Slovenia using a wastewater surveillance system, demonstrating its usefulness as a complementary tool in epidemiological surveillance. This study found that estimated SARS-CoV-2 infections in Slovenia peaked in September 2022 and showed a declining trend with subsequent lower peaks in March–April and December 2023, mirroring the trends observed from clinical data. Based on both surveillance systems, the most prevalent variant in 2022 was BA.5. By 2023, BQ.1 and other Omicron variants increased in prevalence. By the end of 2023, XBB sublineages and the BA.2.86 variant had become predominant, demonstrating consistent dynamic shifts in variant distribution across both monitoring methods. This study found that wastewater surveillance at wastewater treatment plants in Slovenia effectively tracked SARS-CoV-2 infection trends, showing a moderate to strong correlation with clinical data and providing early indications of changes in infection trends and variant emergence. Despite limitations during periods of low virus concentration, the system proved significant in providing early warnings of infection trends and variant emergence, thus enhancing public health response capabilities.