BACKGROUND: Pakistan is one of the two countries endemic for wild poliovirus type 1 (WPV1). Active clinical and environmental wastewater surveillance along with laboratory investigation is an integral and primary component of the polio eradication strategies. The current study is mainly focused on the virological data to understand the current epidemiology of WPV1 in Pakistan during 2019-2022.
METHODS: 141,037 stool specimens of patients reported with acute flaccid paralysis (AFP) and 3,171 wastewater samples were tested for poliovirus detection using cell culture and PCR. Phylogenetic analysis of WPV1 was performed using MEGA and Nextstrain.
RESULTS: Poliovirus isolates were classified into 15 distinct genetic clusters with multiple transmission lineages. Spatio-temporal trends indicated a significant decline in the incidence of poliomyelitis reported in 58 districts in 2019 to just 3 in 2022. The historical reservoirs in Peshawar, Quetta, and Karachi successfully eliminated the indigenous transmission chains of wild poliovirus active there for years.
CONCLUSIONS: Our findings reinforce the evolving epidemiology of poliovirus in Pakistan which is now confined to South KP. All historically known reservoirs in Peshawar, Karachi and Quetta blocs are now free of poliovirus. Intensified clinical and environmental surveillance should be maintained to eliminate the very few remaining transmission lineages and certify the poliovirus eradication by 2026.

National opinions on a wide variety of public health topics can change over time and have highly contextual nuances. This study is a follow-up to prior inquiries into the knowledge of wastewater-based epidemiology, privacy concerns surrounding sample collection, and the use of data acquired, along with privacy awareness from an online survey conducted in the metropolitan United States during the winter of 2023. Mentions of wastewater-surveillance-related terms in the media remained common. Towards the outbreak tail in 2023, public support for surveillance of toxins (91%), diseases (91%), terrorist threats (87%), illicit drugs (70%), prescription medications (69%), and gun residue (60%) remained high. There was less support for surveillance of alcohol consumption (49%), mental illness (46%), healthy eating (37%), and lifestyle behaviors (35%). In terms of geographic scale, most respondents supported citywide surveillance (85%) with markedly lower levels of support for smaller (less anonymous) geographic scales covered by specific locations. Wastewater surveillance does not receive the public pushback that other COVID-19-related health system actors have witnessed. Instead, the public supports the expansion of wastewater surveillance as a standard to complement public health tools in other areas of health protection.

We conducted environmental surveillance to detect avian influenza viruses circulating at live poultry markets (LPMs) and poultry farms in Guangxi Autonomous Region, China, where near the China-Vietnam border. From November through April 2017-2018 and 2018-2019, we collected environmental samples from 14 LPMs, 4 poultry farms, and 5 households with backyard poultry in two counties of Guangxi and tested for avian influenza A, H5, H7, and H9 by real-time reverse transcription-polymerase chain reaction (rRT-PCR). In addition, we conducted four cross-sectional questionnaire surveys among stall owners on biosecurity practices in LPMs of two study sites. Among 16,713 environmental specimens collected and tested, the median weekly positive rate for avian influenza A was 53.6% (range = 33.5% - 66.0%), including 25.2% for H9, 4.9% for H5, and 21.2% for other avian influenza viruses A subtypes, whereas a total of two H7 positive samples were detected. Among the 189 LPM stalls investigated, most stall owners (73.0%) sold chickens and ducks. Therefore, continued surveillance of the avian influenza virus is necessary for detecting and responding to emerging trends in avian influenza virus epidemiology.

BACKGROUND: Poliovirus is a highly infectious enterovirus (EV) that primarily affects children and can lead to lifelong paralysis or even death. Vaccine-derived polioviruses (VDPVs) are a great threat since they are derived from the attenuated virus in the Oral Poliovirus Vaccine (OPV) and can mutate to a more virulent form. The purpose of this study was to identify VDPV serotype 2 through the year 2020-2021 via surveillance of sewage samples collected from different localities and governorates in Egypt and stool specimens from Acute Flaccid Paralysis (AFP) cases. Both were collected through the national poliovirus surveillance system and according to the guidelines recommended by the WHO.
METHODS: A total of 1266 sewage samples and 3241 stool samples from January 2020 to December 2021 were investigated in the lab according to World Health Organization (WHO) protocol for the presence of Polioviruses by cell culture, molecular identification of positive isolates on L20B cell line was carried out using real-time polymerase chain reactions (RT-PCR). Any positive isolates for Poliovirus type 2 and isolates suspected of Vaccine Derived Poliovirus Type 1 and type 3 screened by (VDPV1) or Vaccine Poliovirus Type 3 (VDPV3) assay in RT-PCR were referred for VP1 genetic sequencing.
RESULTS: The outbreak was caused by circulating VDPV2 (cVDPV2) strains started in January 2021. By the end of February 2021, a total of 11 cVDPV2s were detected in sewage samples from six governorates confirming the outbreak situation. One additional cVDPV2 was detected later in the sewage sample from Qena (June 2021). The first and only re-emergence of VDPV2 in stool samples during the outbreak was in contact with Luxor in June 2021. By November 2021, a total of 80 VDPVs were detected. The Egyptian Ministry of Health and Population (MOHP), in collaboration with the WHO, responded quickly by launching two massive vaccination campaigns targeting children under the age of five. Additionally, surveillance systems were strengthened to detect new cases and prevent further spread of the virus.
CONCLUSIONS: The continued threat of poliovirus and VDPVs requires ongoing efforts to prevent their emergence and spread. Strategies such as improving immunization coverage, using genetically stable vaccines, and establishing surveillance systems are critical to achieving global eradication of poliovirus and efficient monitoring of VDPVs outbreaks.

Wastewater-based epidemiology (WBE) is an environmental approach to monitor community health through the analysis of sewage. The COVID-19 pandemic catalyzed scientists and public health professionals to revisit WBE as a tool to optimize resource allocation to mitigate disease spread and prevent outbreaks. Some studies have highlighted the value of WBE programs that coordinate with public health professionals; however, the details necessary for implementation are not well-characterized. To respond to this knowledge gap, this article documents the framework of a successful WBE program in Arizona, titled Wastewater Analysis for Tactical Epidemiological Response Systems (WATERS), detailing the developed structure and methods of communication that enabled public health preparedness and response actions. This communication illustrates how program operations were employed to reduce outbreak severity. The structure outlined here is customizable and may guide other programs in the implementation of WBE as a public health tool.

Digital polymerase chain reaction (dPCR) has emerged as a groundbreaking technology in molecular biology and diagnostics, offering exceptional precision and sensitivity in nucleic acid detection and quantification. This review highlights the core principles and transformative potential of dPCR, particularly in infectious disease diagnostics and environmental surveillance. Emphasizing its evolution from traditional PCR, dPCR provides accurate absolute quantification of target nucleic acids through advanced partitioning techniques. The review addresses the significant impact of dPCR in sepsis diagnosis and management, showcasing its superior sensitivity and specificity in early pathogen detection and identification of drug-resistant genes. Despite its advantages, challenges such as optimization of experimental conditions, standardization of data analysis workflows, and high costs are discussed. Furthermore, we compare various commercially available dPCR platforms, detailing their features and applications in clinical and research settings. Additionally, the review explores dPCR\'s role in water microbiology, particularly in wastewater surveillance and monitoring of waterborne pathogens, underscoring its importance in public health protection. In conclusion, future prospects of dPCR, including methodological optimization, integration with innovative technologies, and expansion into new sectors like metagenomics, are explored.

Escherichia coli, both commensal and pathogenic, can colonize plants and persist in various environments. It indicates fecal contamination in water and food and serves as a marker of antimicrobial resistance. In this context, 61 extended-spectrum β-lactamase (ESBL)-producing E. coli from irrigation water and fresh produce from previous studies were characterized using whole genome sequencing (Illumina MiSeq). The Center for Genomic Epidemiology and Galaxy platforms were used to determine antimicrobial resistance genes, virulence genes, plasmid typing, mobile genetic elements, multilocus sequence typing (MLST), and pathogenicity prediction. In total, 19 known MLST groups were detected among the 61 isolates. Phylogroup B1 (ST58) and Phylogroup E (ST9583) were the most common sequence types. The six ST10 (serotype O101:H9) isolates carried the most resistance genes, spanning eight antibiotic classes. Overall, 95.1% of the isolates carried resistance genes from three or more classes. The blaCTX-M-1, blaCTX-M-14, and blaCTX-M-15 ESBL genes were associated with mobile genetic elements, and all of the E. coli isolates showed a >90% predicted probability of being a human pathogen. This study provided novel genomic information on environmental multidrug-resistant ESBL-producing E. coli from fresh produce and irrigation water, highlighting the environment as a reservoir for multidrug-resistant strains and emphasizing the need for ongoing pathogen surveillance within a One Health context.

In regions without adequate centralized wastewater treatment plants, sample collection from rivers and sewers can be an alternative sampling strategy for wastewater surveillance. This study aimed to assess the feasibility of alternative sampling strategies by testing samples collected from rivers (n = 246) and sewers (n = 244) in the Kathmandu Valley between March 2021 and February 2022. All samples were concentrated using the skimmed-milk flocculation method and severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA was quantified using the nucleocapsid (N) and envelope (E) genes qPCR assays. Of the total, 75 % (371/490) of the samples tested positive using at least one qPCR assay, with concentrations ranging from 3.0 to 8.3 log10 gene copies/L. No significant correlation between concentrations of SARS-CoV-2 from both sewers and river with the number of confirmed coronavirus disease 2019 (COVID-19) cases in the Kathmandu valley was observed (p > 0.05). Despite the high concentration of SARS-CoV-2 in rivers and sewers, we hypothesize this finding to be a result of inaccurate number of clinical cases possibly due to inadequate clinical testing. This longitudinal study further supports the statement to consider sampling strategies from sewers and rivers for WBS in Nepal and other low and middle-income countries.

Precise and rapid methods are needed to improve monitoring approaches of L. pneumophila (Lp) in cooling towers (CTs) to allow timely operational adjustments and prevent outbreaks. The performance of liquid culture (ASTM D8429-21) and an online qPCR device were first compared to conventional filter plate culture (ISO 11731-2017), qPCR and semi-automated qPCR at three spiked concentrations of Lp (serogroup 1) validated by flow cytometry (total/viable cell count). The most accurate was qPCR, followed by liquid culture, online and semi-automated qPCR, and lastly, by a significant margin, filter plate culture. An industrial CT system was monitored using liquid and direct plate culture by the facility, qPCR and online qPCR. Direct plate and liquid culture results agreed at regulatory sampling point, supporting the use of the faster liquid culture for monitoring culturable Lp. During initial operation, qPCR and online qPCR results were within one log of culture at the primary pump before deviating after first cleaning. Other points revealed high spatial variability of Lp. The secondary pumps and chiller had the most positivity and highest concentrations by both qPCR and liquid culture compared to the basin and infeed tank. Altogether, this suggests that results from monthly compliance sampling at a single location with plate culture are not representative of Lp risks in this CT due to the high temporal and spatial variability. The primary pump, rather than the CT basin, should be designated for sampling, as it is representative of the health risk. An annual multi point survey of the system should be conducted to identify and target Lp hot spots. Generally, a combination of liquid culture for compliance and frequent qPCR for process control provides a more agile and robust monitoring scheme than plate culture alone, enabling early treatment adjustments, due to lower limit of detection (LOD) and turnover time.

After the first phase of the COVID-19 pandemic in Europe, a new highly pathogenic variant of echovirus 11 (E11) was detected. The aim of this study was to analyze the genetic diversity of Polish E11 environmental and clinical strains circulating between 2017 and 2023 as well as compare them with E11 strains isolated from severe neonatal sepsis cases reported in Europe between 2022 and 2023. Additionally, the study explores the effectiveness of environmental monitoring in tracking the spread of new variants. For this purpose, the complete sequences of the VP1 capsid protein gene were determined for 266 E11 strains isolated in Poland from 2017 to 2023, and phylogenetic analysis was performed. In the years 2017-2023, a significant increase in the detection of E11 strains was observed in both environmental and clinical samples in Poland. The Polish E11 strains represented three different genotypes, C3, D5 and E, and were characterized by a high diversity. In Poland, the intensive circulation of the new variant E11, responsible for severe neonatal infections with a high mortality in Europe, was detected in the years 2022-2023. This investigation demonstrates the important role of environmental surveillance in the tracking of enteroviruses circulation, especially in settings with limited clinical surveillance.