Antibiotic-Resistant Bacteria and Resistance Genes in Isolates from Ghanaian Drinking Water Sources.

The control of infectious diseases is seriously threatened by the increase in the number of microorganisms resistant to antimicrobial agents. Antibiotic-resistant bacteria have also been identified in the water environment. A field study was performed sampling drinking water sources in seven districts of southern Ghana targeting boreholes, dams, hand-dug wells, and streams during baseflow conditions. Bacteria were isolated (N = 110) from a total of 67 water samples to investigate their antimicrobial susceptibility and to determine their carriage of select antibiotic resistance genes. Bacterial identification was performed using conventional selective media methods and the analytical profile index (API) method. Antibiotic susceptibility tests were carried out using the Kirby-Bauer method. Results indicated that all water sources tested were of poor quality based on the presence of fecal indicator organisms. The most commonly occurring bacterium isolated from water was Klebsiella spp. (N = 24, 21.8%), followed by E. coli (N = 23, 20.9%). Gram-negative bacteria isolates were most commonly resistant to cefuroxime (24.5%), while the Gram-positives were most commonly resistant to meropenem (21.3%). The highest rates of bacterial resistances to more than one antibiotic were observed in Klebsiella spp. (30.0%) followed by E. coli (27.8%). PCR was used to detect the presence of a select antibiotic resistance genes in the Gram-negative isolates. The presence of bla NDM-1, sull, tet(O), and tet(W) were observed in isolates from all water sources. In contrast, ermF was not detected in any of the Gram-negative isolates from any water source. Most (28.7%) of the resistance genes were observed in E. coli isolates. Reducing microbial contamination of the various water sources is needed to protect public health and to ensure the sustainability of this resource. This further calls for education of the citizenry.

Year-long wastewater monitoring for SARS-CoV-2 signals in combined and separate sanitary sewers.

COVID-19 wastewater-based epidemiology has been performed in catchments of various sizes and sewer types with many short-term studies available and multi-seasonal studies emerging. The objective of this study was to compare weekly observations of SARS-CoV-2 genes in municipal wastewater across multiple seasons for different systems as a factor of sewer type (combined, separate sanitary) and system size. Sampling occurred following the first wave of SARS-CoV-2 cases in the study region (June 2020) and continued through the third wave (May 2021), the period during which clinical testing was widely available and different variants dominated clinical cases. The strongest correlations were observed between wastewater N1 concentrations and the cumulative clinical cases reported in the 2 weeks prior to wastewater sampling, followed by the week prior, new cases, and the week after wastewater sampling. Sewer type and size did not necessarily explain the strength of the correlations, indicating that other non-sewer factors may be impacting the observations. In-system sampling results for the largest system sampled are presented for 1 month. Removing wet weather days from the data sets improved even the flow-normalized correlations for the systems, potentially indicating that interpreting results during wet weather events may be more complicated than simply accounting for dilution. PRACTITIONER POINTS: SARS-CoV-2 in wastewater correlated best with total clinical cases reported in 2 weeks before wastewater sampling at the utility level. Study performed when clinical testing was widespread during the year after the first COVID-19 wave in the region. Sewer type and size did not necessarily explain correlation strength between clinical cases and wastewater-based epidemiology results. Removing wet weather days improved correlations for 3/4 utilities studied, including both separate sanitary and combined sewers.

Metabolically Active Prokaryotes and Actively Transcribed Antibiotic Resistance Genes in Sewer Systems: Implications for Public Health and Microbially Induced Corrosion.

Sewer systems are reservoirs of pathogens and bacteria carrying antibiotic resistance genes (ARGs). However, most recent high-throughput studies rely on DNA-based techniques that cannot provide information on the physiological state of the cells nor expression of ARGs. In this study, wastewater and sewer sediment samples were collected from combined and separate sanitary sewer systems. The metabolically active prokaryote community was evaluated using 16S rRNA amplicon sequencing and actively transcribed ARG abundance was measured using mRNA RT-qPCR. Three (sul1, blaTEM, tet(G)) of the eight tested ARGs were quantifiable in select samples. Sewer sediment samples had greater abundance of actively transcribed ARGs compared to wastewater. Microbiome analysis showed the presence of metabolically active family taxa that contain clinically relevant pathogens (Pseudomonadaceae, Enterobacteraceae, Streptococcaceae, Arcobacteraceae, and Clostridiaceae) and corrosion-causing prokaryotes (Desulfobulbaceae and Desulfovibrionaceae) in both matrices. Spirochaetaceae and methanogens were more common in the sediment matrix while Mycobacteraceae were more common in wastewater. The microbiome obtained from 16S rRNA sequencing had a significantly different structure from the 16S rRNA gene microbiome. Overall, this study demonstrates active transcription of ARGs in sewer systems and provides insight into the abundance and physiological state of taxa of interest in the different sewer matrices and sewer types relevant for wastewater-based epidemiology, corrosion, and understanding the hazard posed by different matrices during sewer overflows.

Comparison of residential dormitory COVID-19 monitoring via weekly saliva testing and sewage monitoring.

Wastewater surveillance has been used as a tool for COVID-19 outbreak detection particularly where there was not capability in place for routine and robust individual testing. Given clinical reports that earlier detection is possible following infection from throat/nasal samples compared to fecal samples for COVID-19 patients, the utility of wastewater testing where robust individual testing is possible is less clear. The objective of this study was to compare the results of weekly required COVID-19 saliva tests to weekly wastewater monitoring for residential buildings (i.e., dormitories) located across three college campuses capturing wastewater from 80 to 441 occupants per sampling location. Sampling occurred during the spring semester of the 2021 academic year which captured the third wave of SARS-CoV-2 cases in the study region. Comparison of the saliva and wastewater testing results indicated that the wastewater SARS-CoV-2 concentrations had a strong linear correlation with the previous week's percentage of positive saliva test results and a weak linear correlation with the saliva testing results surrounding the wastewater sampling (four days before and 3 days after). Given that no correlation was observed between the wastewater and the saliva testing from the following week, the weekly saliva testing captured spikes in COVID-19 cases earlier than the weekly wastewater sampling. Interestingly, the N1 gene was observed in buildings on all campuses, but N2 was observed in wastewater on only one of the campuses. N1 and N2 were also observed in sewer biofilm. The campus-specific challenges associated with implementation of wastewater surveillance are discussed. Overall, these results can help inform design of surveillance for early detection of SARS-CoV-2 in residential settings thereby informing mitigation strategies to slow or prevent the spread of the virus among residents in congregate living.

Viability-based quantification of antibiotic resistance genes and human fecal markers in wastewater effluent and receiving waters.

Antibiotic resistance is a public health issue with links to environmental sources of antibiotic resistance genes (ARGs). ARGs from nonviable sources may pose a hazard given the potential for transformation whereas ARGs in viable sources may proliferate during host growth or conjugation. In this study, ARGs in the effluent from three municipal wastewater treatment plants (WWTPs) and the receiving surface waters were investigated using a viability-based qPCR technique (vPCR) with propidium monoazide (PMA). ARGs sul1, tet(G), and blaTEM, fecal indicator marker BacHum, and 16S rRNA gene copies/mL were found to be significantly lower in viable-cells than in total concentrations for WWTP effluent. Viable-cell and total gene copy concentrations were similar in downstream samples except for tet(G). Differences with respect to season in the prevalence of nonviable ARGs in surface water or WWTP effluent were not observed. The results of this study indicate that qPCR may overestimate viable-cell ARGs and fecal indicator genes in WWTP effluent but not necessarily in the surface water >1.8 km downstream.