To improve the efficiency of phosphorus (P) release from marine sediments and contribute to P loop closure, this study proposed a novel strategy combining bio-inoculation with polyphosphate-accumulating organisms (PAOs) and chemical enhancement via chelating agents. Based on prior findings, two-stage experiments were conducted. In Stage 1, anaerobic batch tests assessed the effect of different chelating agents for P release. While citrate showed no promoting effect, the addition of ethylenediaminetetraacetic acid (EDTA) significantly enhanced total P release, reaching 48.5 % within 15 days. In Stage 2, PAO-acclimated sediments were introduced into the system, followed by alternating anaerobic-aerobic fed-batch operation for 7 days, and subsequent EDTA addition with anaerobic incubation for another 6 days. This combined approach achieved a total P release efficiency 83.4 %, with final soluble P concentrations reaching 145.9 mg/L. During this process, PAOs were rapidly enriched, with their relative abundance increasing from 12.9 % to 65.0 %. Metagenomic analysis revealed that EDTA promoted environmental filtering, selectively enriching PAOs (Candidatus Accumulibacter) and thereby reinforcing their specific contributions to P functional genes. The resulting P-rich supernatant was then subjected to precipitation. PHREEQC simulations guided the prediction of optimal precipitation conditions, and laboratory experiments confirmed that most soluble P, especially Fe-bound forms, could be efficiently recovered, with maximum precipitation efficiencies of 98.8 %.

Wastewater-based surveillance (WBS) allows the analysis of pathogens, chemicals or other biomarkers in wastewater to derive unbiased epidemiological information at population scale. After re-gaining attention during the SARS-CoV-2 pandemic, the field holds promise as a surveillance and early warning system by tracking emerging pathogens with pandemic potential. Expanding the current toolbox of analytical techniques for wastewater analysis, we explored the use of Hyperplex PCR (hpPCR) to analyse SARS-CoV-2 mutations in wastewater samples collected weekly in up to 22 sites across Sweden between October 2022 and December 2023. The samples were tested using a probe panel ranging from 10- to 18-plex, continuously adapted within 1–2 weeks to quantify relevant mutations of concern over time. For cross-validation, the samples were simultaneously analysed with commonly used methods including quantitative PCR (qPCR) and next-generation sequencing (NGS). hpPCR is demonstrated herein to provide (1) systematic single nucleotide specificity with a straightforward probe design, (2) high multiplexity with minimal panel re-optimization requirements and (3) 4–5-week earlier mutation detection relative to NGS with comparable performance of mutation frequency quantification (Pearson r = 0.88, n = 50). Hence, hpPCR is shown to be a powerful complementary tool to the current workflow involving NGS and qPCR by facilitating the assembly of dynamic high-plex panels compatible with high-frequency monitoring of multiple key pathogens and/or variants in WBS.

Volatile fatty acids (VFAs) are platform chemicals with a higher value with wide applications. The feasibility of producing high-purity VFAs from source-separated wastewater using mixed-culture fermentation and membrane techniques was investigated. Batch studies were conducted for VFA production from blackwater and food waste under acidic and alkaline conditions. The VFA production from blackwater was higher at initial pH 9 with yield of up to 0.82 ± 0.03 gCOD/gCOD_fed due to higher buffer capacity, homogeneity, and biodegradability. The highest VFA yield from food waste was 0.36 ± 0.02 gCOD/gCOD_fed at initial pH 5. VFAs from the blackwater were dominated by acetic acid (up to 93 %), regardless of pH VFAs from the food waste were dominated by butyric acid (up to 76 %) and propionic acid (up to 52 %) at pH 5 and 9, respectively. Both the substrate types and pH influenced the microbial communities of the fermentation reactors. Bacteroides (up to 40 %) and Atopostipes (up to 51 %) were dominant genii for blackwater at initial pH 5 and 9, while Clostridium_sensu_stricto clusters (up to 58 %) and Romboutsia (up to 37 %) dominated food waste fermentation microbial communities at pH 5 and 9, respectively. VFAs were separated and purified with nanofiltration (NF) and reverse osmosis (RO). NF produced high-purity, but low-concentration permeate (900 −1500 mgCOD/L) at 50 % permeate recovery (with up to 42 % of acetic acid permeating the NF membrane). A higher concentration of pure VFAs (3x higher) was achieved with a subsequent step of RO. The study highlights the feasibility of the recovery of functional chemicals from waste.

Wastewater monitoring is highly efficient in SARS-CoV-2 surveillance for tracking virus spread through travel, surpassing traditional airport passenger testing. This study explored the links between SARS-CoV-2 contents and variants from aircraft to city, assessing the impact of detected variants from international travellers versus the local population. A total of 969 variants using next-generation sequencing (NGS) were examined to understand the links between—aircraft, Arlanda airport, wastewater treatment plants (WWTPs), and Stockholm city—and compared these to variants detected in Stockholm hospitals from January to May 2023. SARS-CoV-2 contents in WWTPs reflected local infection rates, requiring analysis from multiple plants for an accurate city-wide infection assessment. Variants initially detected in aircraft arriving from China did not spread widely during the study period. RT-qPCR is adequate for the detection of specific variants in wastewater, including Variants Under Monitoring. However, NGS remains a powerful method for identifying novel variants. Wastewater monitoring was more effective than clinical testing in the early detection of specific variants, with notable delays observed in clinical surveillance. Furthermore, a broad range of variants are detected in wastewater that surpasses clinical tests. This underscores the vital role of wastewater-based epidemiology in managing future outbreaks and enhancing global health security.

Human adenovirus (HAdV) type F41 has been identified as a possible cause of the non-A-to-E hepatitis outbreak. This study uses wastewater monitoring to track HAdV F40 and F41, supporting clinical investigations and providing insights into the pathogen's role in the outbreak. Given the limited clinical monitoring in Sweden of HAdV-F40/41, this approach also helps estimate the true infection burden of this pathogen during the outbreak. This study developed three qPCR assays for the hexon, penton, and fiber genes of HAdV F40 and F41. The hexon assay was F41-specific, while the fiber assay detected multiple HAdV-F strains. Comprehensive monitoring of HAdV-F40/41 levels in Stockholm's wastewater was conducted over 1.5 years, capturing the period before, during, and after the outbreak. A significant infection wave was observed in spring 2022, with strains beyond lineage 2 contributing to the outbreak. Moreover, simultaneous SARS-CoV-2 surveillance revealed that HAdV-F infections peaked at different times from COVID-19, but the HAdV-F wave aligned with the relaxation of pandemic restrictions. These findings offer valuable insights for future HAdV-F investigations and confirm its role in the non-A-to-E hepatitis outbreak.

Global usage of pharmaceuticals has led to the proliferation of bacteria that are resistant to antimicrobial treatments, creating a substantial public health challenge. Here, we investigate the emergence of sulfonamide resistance genes in groundwater and surface water in Patna, a rapidly developing city in Bihar, India. We report the first quantification of three sulfonamide resistance genes (sulI, sulII and sulIII) in groundwater (12-107 m in depth) in India. The mean relative abundance of gene copies was found to be sulI (2.4 x 10(-2) copies/16S rRNA gene) > sulII (5.4 x 10(-3) copies/16S rRNA gene) > sulIII (2.4 x 10(-3) copies/16S rRNA gene) in groundwater (n = 15) and surface water (n = 3). A comparison between antimicrobial resistance (AMR) genes and wastewater indicators, particularly tryptophan:fulvic-like fluorescence, suggests that wastewater was associated with AMR gene prevalence. Urban drainage channels, containing hospital and domestic wastes, are likely a substantial source of antimicrobial resistance in groundwater and surface water, including the Ganges (Ganga) River. This study is a reference point for decision-makers in the fight against antimicrobial resistance because it quantifies and determines potential sources of AMR genes in Indian groundwater.

This study examined the anaerobic release of phosphorus (P) from two different Baltic Sea sediments (B and F), focusing on the impact of initial concentration of externally introduced waste-derived volatile fatty acids (VFA) as the carbon source, temperature, pH, and mixing conditions. The first batch bioreactor set was operated to demonstrate the effect of VFA on anaerobic P release at different concentrations (1000–10000 mg/L as COD) at 20 °C. A notable P release of up to 15.85 mg/L PO4–P was observed for Sediment B at an initial carbon concentration of 10000 mg COD/L. However, VFA consumption in the bioreactors was minimal or no subsequent. The second batch bioreactor set was carried out to investigate the effect of temperature (20 °C-35 °C), pH (5.5, 7.0 and 8.5) and mixing conditions on P release by introducing lower initial carbon concentration (1000 mg COD/L) considering the potential risk for VFA accumulation in the bioreactors. Maximum P releases of 4.4 mg/L and 3.5 mg/L were for Sediment B and Sediment F, respectively. Two-way ANOVA tests revealed that the operation time and pH and their interactions were statistically significant (p < 0.05) for both sediments while the effect of mixing was not statistically significant. Most of the sulfate was reduced during batch bioreactor operation and Desulfomicobiaceae became dominant among other sulfate-reducing bacteria (SRB) possibly shows the importance of SRB in terms of anaerobic P release. This study gives an insight into future implementations of phosphorus mining from eutrophic environment under anaerobic conditions.

To evaluate the feasibility of enriching polyphosphate-accumulating organisms (PAOs) within marine sediment for achieving phosphorus (P) recovery, two sediment-inoculated sequencing batch reactors (SBRs), fed with propionic acid (R1) and glucose (R2), were operated for 119 days. For comparison, two sewage sludge-inoculated reactors (R3 and R4) were also set up. The sediments/sludge fed with 200 mg/L chemical oxygen demand (COD) equivalent propionic acid exhibited satisfactory P release/uptake performance after 56 days of culture. The maximum P release and uptake rates for R1 were 3 mg P/g VSS•h−1 and 2.5 mg P/g VSS•h−1, respectively, while for R3 they were 2.6 mg P/g VSS•h−1 and 5.8 mg P/g VSS•h−1, respectively. Meanwhile, the PAO family (Rhodocyclaceae) in R1 increased from almost 0 % initially to 16.0 % after 42 days. However, the glucose-fed SBRs did not exhibit enhanced biological phosphorus removal (EBPR) performance throughout the operation. As the COD feeding concentration increased to 400 mg/L, the reactors showed EBPR deterioration. Total P in R1 and R3 significantly decreased from 423.7 mg to 307.2 mg and from 368.0 mg to 94.9 mg, respectively. Key intracellular polymer responses indicated that introduction of excessively high COD significantly reduced poly-P content and the anaerobic synthesis of polyhydroxyalkanoate. Microbial analysis suggested that the breakdown of EBPR performance could be attributed to glycogen-accumulating organisms outcompeting PAOs under high carbon feeding conditions. Additionally, PHREEQC simulations confirmed that P-rich supernatant from the anaerobic phase could theoretically be recovered as struvite, with a recovery efficiency of up to 94 %.

The study of how micro-organisms detect and respond to different stresses has a long history of producing fundamental biological insights while being simultaneously of significance in many applied microbiological fields including infection, food and drink manufacture, and industrial and environmental biotechnology. This is well-illustrated by the large body of work on acid stress. Numerous different methods have been used to understand the impacts of low pH on growth and survival of micro-organisms, ranging from studies of single cells to large and heterogeneous populations, from the molecular or biophysical to the computational, and from well-understood model organisms to poorly defined and complex microbial consortia. Much is to be gained from an increased general awareness of these methods, and so the present review looks at examples of the different methods that have been used to study acid resistance, acid tolerance, and acid stress responses, and the insights they can lead to, as well as some of the problems involved in using them. We hope this will be of interest both within and well beyond the acid stress research community.

To investigate the possibility of phosphorus (P) recovery from marine sediment and explore the role of the carbon: nitrogen ratio in affecting the internal P release under anaerobic conditions, we experimented with the external addition of carbon (acetic acid and glucose) and ammonia nitrogen (NH4-N) to expose P release mechanisms. The 24-day anaerobic incubations were conducted with four different carbon: nitrogen dosing groups including no NH4-N addition and COD/N ratios of 100, 50, and 10. The P release showed that extra NH4-N loading significantly suppressed the decomposition of P (p < 0.05) from the marine sediment, the maximum P release was 4.07 mg/L and 7.14 mg/L in acetic acid- and glucose-fed systems, respectively, without extra NH4-N addition. Additionally, the results exhibited that the imbalance of carbon: nitrogen not only failed to induce the production of organic P mineralization enzyme (alkaline phosphatase) in the sediment but also suppressed its activity under anaerobic conditions. The highest enzyme activity was observed in the group without additional NH4-N dosage, with rates of 1046.4 mg/(kg∙h) in the acetic acid- and 967.8 mg/(kg∙h) in the glucose-fed system, respectively. Microbial data analysis indicated that a decrease in the abundance of P release-regulating bacteria, including polyphosphate-accumulating organisms (Rhodobacteraceae) and sulfate-reducing bacteria (Desulfosarcinaceae), was observed in the high NH4-N addition groups. The observed reduction in enzyme activity and suppression of microbial activity mentioned above could potentially account for the inhibited P decomposition in the presence of high NH4-N addition under anaerobic conditions. The produced P-enriched solution from the bioreactors may offer a promising source for future recovery endeavors.