Environmental Samples Research Articles

Expanding the Diversity of Actinobacterial Tectiviridae: A Novel Genus from Microbacterium

Six novel Microbacterium phages belonging to the Tectiviridae family were isolated using Microbacterium testaceum as a host. Phages MuffinTheCat, Badulia, DesireeRose, Bee17, SCoupsA, and LuzDeMundo were purified from environmental samples by students participating in the Science Education Alliance Phage Hunters Advancing Genomics and Evolutionary Science (SEA-PHAGES) program at Alliance University, New York. The phages have linear dsDNA genomes 15,438–15,636 bp with 112–120 bp inverted terminal repeats. Transmission electron microscopy (TEM) imaging analysis revealed that the six novel phages have six-sided icosahedral double-layered capsids with an internal lipid membrane that occasionally forms protruding nanotubules. Annotation analysis determined that the novel Microbacterium phages all have 32–34 protein-coding genes and no tRNAs. Like other Tectiviridae, the phage genomes are arranged into two segments and include three highly conserved family genes that encode a DNA polymerase, double jelly-roll major capsid protein, and packaging ATPase. Although the novel bacteriophages have 91.6 to 97.5% nucleotide sequence similarity to each other, they are at most 58% similar to previously characterized Tectiviridae genera. Consequently, these novel Microbacterium phages expand the diversity of the Tectiviridae family, and we propose they form the sixth genus, Zetatectivirus.

MeStanG—Resource for High-Throughput Sequencing Standard Data Sets Generation for Bioinformatic Methods Evaluation and Validation

Metagenomics analysis has enabled the measurement of the microbiome diversity in environmental samples without prior targeted enrichment. Functional and phylogenetic studies based on microbial diversity retrieved using HTS platforms have advanced from detecting known organisms and discovering unknown species to applications in disease diagnostics. Robust validation processes are essential for test reliability, requiring standard samples and databases deriving from real samples and in silico generated artificial controls. We propose a MeStanG as a resource for generating HTS Nanopore data sets to evaluate present and emerging bioinformatics pipelines. MeStanG allows samples to be designed with user-defined organism abundances expressed as number of reads, reference sequences, and predetermined or custom errors by sequencing profiles. The simulator pipeline was evaluated by analyzing its output mock metagenomic samples containing known read abundances using read mapping, genome assembly, and taxonomic classification on three scenarios: a bacterial community composed of nine different organisms, samples resembling pathogen-infected wheat plants, and a viral pathogen serial dilution sampling. The evaluation was able to report consistently the same organisms, and their read abundances as provided in the mock metagenomic sample design. Based on this performance and its novel capacity of generating exact number of reads, MeStanG can be used by scientists to develop mock metagenomic samples (artificial HTS data sets) to assess the diagnostic performance metrics of bioinformatic pipelines, allowing the user to choose predetermined or customized models for research and training.

Adapting Methods for Isolation and Enumeration of Microplastics to Quantify Tire Road Wear Particles with Confirmation by Pyrolysis GC-MS.

The complex, varied composition (i.e., rubbers/elastomers, carbon black, fillers, additives, and embedded road materials) and wide density range of tire road wear particles (TRWPs) present challenges for their isolation and identification from environmental matrices. Reliable quantification is important for understanding the environmental fate and potential adverse effects of TRWPs. To address environmental monitoring needs, the present work adapts a series of isolation and identification steps from methods commonly applied for microplastic analysis for single-particle-level enumeration of TRWPs from environmental samples. We present the method performance of a two-stage density separation with saturated NaCl and sodium polytungstate to isolate TRWPs from sediment matrices, the compatibility of tire microrubber with reagents used for digestion of environmental matrices, and the use of elasticity and heat resistance observations to differentiate TRWPs from bitumen particulates, which are potential visual interference. We found that alkaline digestions (NaOH and KOH) are compatible with tire microrubbers, but hydrogen peroxide and sodium hypochlorite can cause TRWPs to lose elasticity and flatten or break when probed. The adapted methods were applied to road dust samples, and a subset of identified TRWPs was qualitatively confirmed by both scanning electron microscopy and pyrolysis-gas chromatography-mass spectrometry. Further, a compilation of aspect ratio measurements of TRWPs between 63 and 500 μm (N = 780) that were isolated from urban sediments shows the potential diversity of TRWP shapes in the environment.

Tb-based Metal-Organic Framework-Referenced Fluorescence Assay for Distinguishing Hydroquinone from Its Isomers and Subsequent Quantitative Visual Detection of Cu2.

Hydroquinone (HQ) and copper ions (Cu2+) are categorized as environmental pollutants that are severely limited in water. Designing a selective assay for discriminating HQ from its two isomers and the convenient determination of Cu2+ is of great importance. Herein, a Tb-based metal-organic framework (Tb-MOF) and HQ are assembled innovatively into a ratiometric fluorescence nanoprobe to selectively distinguish HQ and subsequent quantitative visual detection of Cu2+. The native blue emission of HQ at 338 nm is used as a response signal, while Tb-MOF with green fluorescence offers a reference signal at 545 nm. Notably, neither resorcinol (RC) nor catechol (CC) exhibits obvious emission under the same experimental conditions, which enables discriminating HQ from its isomers. Thus, a ratiometric fluorescence method has been designed for the selective detection of HQ with the fluorescence intensity ratio F338/F545 as the readout. The redox reaction between HQ and Cu2+ induces fluorescence quenching of HQ and no change to that of Tb-MOF, resulting in a noticeable color variation from blue-green to green via the naked eye. Furthermore, sensitive visual detection of Cu2+ is achieved with a low detection limit of 1.67 μM using a smartphone. The satisfactory recoveries and good repeatability of quantitative visualization determined in spiked water samples make this sensing platform suitable for on-site monitoring of environmental samples.

Rapid identification of bacterial select agents using loop-mediated isothermal amplification

BackgroundPoint of need diagnostics provide efficient testing capability for remote or austere locations, decreasing the time to answer by minimizing travel or sample transport requirements. Loop-mediated isothermal amplification (LAMP) is an appealing technology for point-of-need diagnostics due to its rapid analysis time and minimal instrumentation requirements.MethodsHere, we designed and optimized nine LAMP assays that are sensitive and specific to targeted bacterial select agents including Bacillus anthracis, Francisella tularensis, Yersinia pestis, and Brucella spp. Evaluation of each assay determined preliminary limit of detection (LOD) with LOD confirmed across 60 replicates (≥ 95% positivity rate). Testing across a robust set of strains of the target agent, common DNA agents, and near-neighbors documented sensitivity and specificity for independent assays.ResultsSpecifically, all assays were 100% specific and sensitive except for Y. pestis Caf1 (90% inclusive across Y. pestis strains).ConclusionHere, we optimized assay turn-around-time, decreasing a standard 60 min traditional polymerase chain reaction (PCR) to 30 min using LAMP with positive results in as little as 5–10 min. Incorporating point of need sample processing and evaluating the potential inhibitory impact of sample matrices such as whole blood and soil would be needed to enable this test system for use on field-forward clinical and environmental sample testing.

Control of Nanoparticle Size of Intrinsically Fluorescent PET (Polyethylene Terephthalate) Particles Produced Through Nanoprecipitation

Plastics are widely produced due to their stability and ease of manufacturing, but many of them quickly become a waste, breaking down into microplastics and nanoplastics. While methods for the identification and characterization of plastic particles are well consolidated, the small size of nanoplastics presents challenges for their detection and analysis. Furthermore, due to the difficulty of identifying nanoplastics, analytical studies concerning their effect on cells and a comprehensive spectroscopic characterization are still lacking. In this paper, we overcome this obstacle by synthesizing and characterizing, for the first time, PET nanoparticles with specific, stable dimensions through a top-down approach. Using hexafluoroisopropanol-chloroform as a solvent, we prepared PET solutions at various concentrations and analyzed their spectral properties over time. Our results show that PET aggregates into nanoparticles, the quantity of which increases with concentration. These findings provide crucial insights for the detection of nanoplastics in environmental samples through fluorescence measurements and can potentially be used to produce stable PET nanoparticles to evaluate their cytotoxicity.

A Novel High-Conjugated Probe of 4-(Acridone-10-yl)-Phenylethyl Chloroformate (APE-Cl) for Rapid Detection of Amino Compounds Using HPLC with Fluorescence Detection.

The fluorescence detection of amino compounds and the evaluation of their content in environmental samples are vital, not only for assessing food quality but also for studying soil organic matter. Here, we present the synthesis and application of a novel fluorescent probe, 4-(9-acridone)benzylmethyl carbonochloride (APE-Cl), for detecting amino compounds via a chloroformate reaction with fluorescence detection. The complete derivatization reaction of APE-Cl with amino compounds can be accomplished in aqueous acetonitrile within 5min at room temperature, using 0.2M borate buffer (pH = 9.0). APE-amine derivatives exhibited intense fluorescence with an excitation maximum at λex 254nm and an emission maximum at λem 418nm. All derivatives demonstrated high stability, strong fluorescence, and elevated ionization potential under atmospheric pressure chemical ionization (APCI-MS) in positive ion detection mode. The method, combined with gradient elution, provides baseline resolution of common amine derivatives on a reversed-phase C18 column. The LC separation for the derivatized amines shows good reproducibility with aqueous acetonitrile as the mobile phase. The relative standard deviations (n = 6) for each amine derivative are < 3.99%. The detection limits (at a signal-to-noise ratio of 3) per injection ranged from 1.68 to 11.2 femtomole. The established pre-column derivatization method for determining amino compounds in practical samples proved to be satisfactory.

Is green metallic sheen in emb agar accurate to assist the Escherichia coli identification?

Escherichia coli is an essential bacteria as a human and animal pathogen, although this species can also be disseminated in the environment. The clinical laboratories use EMB agar (Eosin Methylene Blue) to identify E. coli from the observation of metallic green sheen. However, other bacterial species may show sheen when sown on EMB agar, or in some cases, the pH sample influences the presence of this sheen by E. coli. In this study, MALDI-TOF was used as the gold standard to detect E. coli from human, animal, and environmental samples by analyzing the presence of green glow on EMB agar. The positive predictive values (PPV) ​​were higher (above 85%) in human, animal, and environmental samples studied, and the negative predictive value (NPV) was variable between the samples. Specificity and sensitivity were also variables suggesting the influence of the pH of the samples in the presence of metallic green brightness. This work demonstrated that it is necessary to implement biochemical tests or, when possible, molecular assays to confirm E. coli since only the observation of the production of metallic green glow in EMB can generate false negative results in the identification of E. coli. Keywords: Biochemical tests; Escherichia coli; Laboratory Analysis; MALDI-TOF.

Green-Emitting AIEgen for Fluorescent Switch-Off Detection of Multi-Cations and Applications.

An easy-to-synthesize aggregation-induced emission (AIE) active Schiff base HNSA was obtained by condensing equimolar amount of 3-hydroxy-2-naphthohydrazide and salicylaldehyde. In pure DMSO, HNSA is non-fluorescent, but increasing the HEPES (H2O, 10 mM, pH 7.4) fraction (fHEPES) ≥ 90% showed an intense green fluorescence with maximum fluorescence intensity at 515nm. The SEM analysis of HNSA at fHEPES 95% supported the formation of sheet-like self-aggregates of HNSA. The HNSA aggregates were found to be emissive over a wide pH range from pH 2.20 to 7.50. Further, metal ions sensing ability by AIE luminogen (AIEgen) HNSA was performed in fHEPES 95%. The fluorescence intensity of AIEgen HNSA was quenched by Co2+, Ni2+, Cu2+ and Fe2+ ions with an estimated detection limit of 0.43 µM, 0.48 µM, 6.26 µM, and 6.74 µM, respectively. AIEgen HNSA coated paper strips were developed for the visual detection of these metal ions. Also, AIEgen HNSA was successfully utilized to qualitatively detect metal ions in chickpea and lima bean sprouts. In addition, AIEgen HNSA was used to prepare green fluorescent gelatin gel and fluorescent invisible ink to write on filter paper to transfer confidential information. The probe AIEgen HNSA was employed to quantify metal ions in real environmental water samples.

A comparison of risk factors for canine leptospirosis and seropositivity in New South Wales, Australia.

This study aimed to identify and compare risk factors associated with canine leptospirosis and Leptospira seropositivity in New South Wales, Australia. Seventy-nine canine cases of confirmed leptospirosis (clinical cases) and 16 healthy dogs seropositive to Leptospira (seropositive cases) were included in the study; these were separately compared to 394 healthy dogs seronegative to Leptospira (controls) in at-risk areas. A questionnaire investigated rat contact, stagnant water, dog park access and household number of dogs and cats. Associations between these factors, signalment and risk of leptospirosis or Leptospira seropositivity were screened using Pearson Chi-Square test and logistic regression was used to estimate odds ratios of association. Significantly associated with both leptospirosis risk and seropositivity risk was rat contact (P < 0.01) which increased the risk 4.3- and 4.5-fold, respectively. Other factors significantly associated with leptospirosis risk were breed (P < 0.01) and age (P < 0.01). Being a herding dog or Terrier increased leptospirosis risk 27.2-fold and 8-fold, respectively and being a young adult dog increased leptospirosis risk 3.9-fold. Frequenting dog parks decreased leptospirosis risk (OR 0.12). In contrast, no other significant risk factors were identified for seropositivity risk. Exposure to Leptospira through contact with reservoir hosts resulted in seropositivity as well as clinical leptospirosis. However, host factors like breed and age seem to be important factors in determining if exposure results in clinical disease. Increased urbanisation and landscape fragmentation could result in increased exposure to reservoir hosts. Environmental sampling is needed to identify sources of infection. Risk mitigation includes reducing contact with reservoir hosts and contaminated environments and increased pest control.

Review of Methods for Studying Viruses in the Environment and Organisms

Recent decades have seen growing attention on viruses in the environment and their potential impacts as a result of global epidemics. Due to the diversity of viral species along with the complexity of environmental and host factors, virus extraction and detection methods have become key for the study of virus ecology. This review systematically summarises the methods for extracting and detecting pathogens from different environmental samples (e.g., soil, water, faeces, air) and biological samples (e.g., plants, animals) in existing studies, comparing their similarities and differences, applicability, as well as the advantages and disadvantages of each method. Additionally, this review discusses future directions for research in this field. The aim is to provide a theoretical foundation and technical reference for virus ecology research, facilitating further exploration and applications in this field.

Emerging biofilm formation and disinfectant susceptibility of ESBL-producing Klebsiella pneumoniae

Klebsiella pneumoniae is an opportunistic pathogen responsible for various infections in humans and animals. It is known for its resistance to multiple antibiotics, particularly through the production of Extended-Spectrum Beta-Lactamases (ESBLs), and its ability to form biofilms that further complicate treatment. This study aimed to isolate and identify K. pneumoniae from animal and environmental samples and assess commercial disinfectants’ effectiveness against K. pneumoniae isolates exhibiting ESBL-mediated resistance and biofilm-forming ability in poultry and equine farms in Giza Governorate, Egypt. A total of 320 samples, including nasal swabs from equine (n = 60) and broiler chickens (n = 90), environmental samples (n = 140), and human hand swabs (n = 30), were collected. K. pneumoniae was isolated using lactose broth enrichment and MacConkey agar, with molecular confirmation via PCR targeting the gyrA and magA genes. PCR also identified ESBL genes (blaTEM, blaSHV, blaCTX-M, blaOXA-1) and biofilm genes (luxS, Uge, mrkD). Antimicrobial susceptibility was assessed, and the efficacy of five commercial disinfectants was evaluated by measuring inhibition zones. Klebsiella pneumoniae was isolated from poultry (13.3%), equine (8.3%), wild birds (15%), water (10%), feed (2%), and human hand swabs (6.6%). ESBL and biofilm genes were detected in the majority of the isolates, with significant phenotypic resistance to multiple antibiotics. The disinfectants containing peracetic acid and hydrogen peroxide were the most effective, producing the largest inhibition zones, while disinfectants based on sodium hypochlorite and isopropanol showed lower efficacy. Statistical analysis revealed significant differences in the effectiveness of disinfectants against K. pneumoniae isolates across various sample origins (P < 0.05). The presence of K. pneumoniae in animal and environmental sources, along with the high prevalence of ESBL-mediated resistance and biofilm-associated virulence genes, underscores the zoonotic potential of this pathogen. The study demonstrated that disinfectants containing peracetic acid and hydrogen peroxide are highly effective against ESBL-producing K. pneumoniae. Implementing appropriate biosecurity measures, including the use of effective disinfectants, is essential for controlling the spread of resistant pathogens in farm environments.

Leptospira biofilms: implications for survival, transmission, and disease management.

Leptospirosis is a zoonotic disease caused by Leptospira bacteria, affecting humans and a broad range of wild and domestic animals in diverse epidemiological settings (rural, urban, and wild). The disease's pathogenesis and epidemiology are complex networks not fully elucidated. Epidemiology reflects the One Health integrated approach of environment-animal-human interaction, causing severe illness in humans and animals, with consequent public health burdens. Saprophytic and pathogenic leptospires have been shown to form biofilms in vivo, in vitro, and in environmental samples. Biofilms are characterized by a polymeric matrix that confers protection against hostile environments (both inside and outside of the host), favoring bacterial survival and dissemination. Despite its significance, the role of this bacterial growth mode in leptospiral survival, transmission, and decreased antibiotic susceptibility remains poorly understood and underexplored. Even so, the literature indicates that biofilms might be correlated with lower antimicrobial susceptibility and chronicity in leptospirosis. In this minireview, we discuss the aspects of biofilm formation by Leptospira and their significance for epidemiology and therapeutic management. Understanding the current scenario provides insight into the future prospects for biofilm diagnosis, prevention, and treatment of leptospirosis.

Ultrasensitive Quantification of Neonicotinoid Thiamethoxam in Environment using MOF-derived CuCo2O4/3D rGO Based Electrochemical Sensor Integrated with Optimized Neural Network.

Accurate quantification of neonicotinoid insecticides is pivotal to ensure environmental safety by examining and mitigating their potential harmful effects on pollinators and aquatic ecosystems. In this scenario, detection of neonicotinoid insecticide, thiamethoxam (TMX), is significant for safeguarding ecological balance and human health. Hence, we developed a highly sensitive electrochemical sensor for detection of TMX in environmental samples, utilizing a novel nanocomposite with superior electrocatalytic properties and integrating an optimized neural network for accurate data analysis. The nanocomposite was synthesized via sonochemical approach, combining metal-organic framework (MOF)-derived spinel copper cobaltite (M-CuCo₂O₄) with three-dimensional reduced graphene oxide (3DrGO). Important characterizations were performed on prepared M-CuCo₂O₄/3DrGO composite and was immobilized on a screen-printed carbon electrode (SPCE) for electrochemical investigations. The synergistic effects of M-CuCo₂O₄ and 3DrGO enabled M-CuCo₂O₄/3DrGO/SPCE to achieve exceptional performance towards TMX detection. The sensor exhibited low limit of detection (LOD) of 0.6 nM and wide linear range of 0.15 to 174.52 μM. Furthermore, neural network model demonstrated excellent accuracy in estimating TMX concentrations, achieving a root mean square error (RMSE) of 2.01 and mean absolute error (MAE) of 1.33. The sensor showed remarkable stability and reliability in real samples including agricultural wastewater, red soil, and brown rice, highlighting its practical applicability for TMX monitoring in environmental and agricultural contexts.

Unipept in 2024: Expanding Metaproteomics Analysis with Support for Missed Cleavages and Semitryptic and Nontryptic Peptides.

Unipept, a pioneering software tool in metaproteomics, has significantly advanced the analysis of complex ecosystems by facilitating both taxonomic and functional insights from environmental samples. From the onset, Unipept's capabilities focused on tryptic peptides, utilizing the predictability and consistency of trypsin digestion to efficiently construct a protein reference database. However, the evolving landscape of proteomics and emerging fields like immunopeptidomics necessitate a more versatile approach that extends beyond the analysis of tryptic peptides. In this article, we present a significant update to the underlying index structure of Unipept, which is now powered by a Sparse Suffix Array index. This advancement enables the analysis of semitryptic peptides, peptides with missed cleavages, and nontryptic peptides such as those encountered in other research fields such as immunopeptidomics (e.g., MHC- and HLA-peptides). This new index benefits all tools in the Unipept ecosystem such as the web application, desktop tool, application programming interface (API), and command line interface. A benchmark study highlights significantly improved performance in handling missed cleavages, preserving the same level of accuracy.

Ribotyping Staphylococcus epidermidis Using Probabilistic Sequence Analysis and Levenshtein Distance Algorithm

Staphylococcus epidermidis (S. epidermidis) live in different human locations and natural environments. For ribotyping S. epidermidis sub-species, 2507 PCR-amplified reads of 16S rRNA genes of S. epidermidis in a public dataset were used for probabilistic sequence analysis. A sequence probability logo (sequence pLogo) as a reference sequence of 16S rRNA genes of S. epidermidis was constructed. Through implementation of Levenshtein Distance algorithm, two 20-base pairs (bp) motifs, commonly present in 2507 PCR-amplified reads, were identified. The top 38 S. epidermidis isolates, which carried 16S rRNA nucleotide domains that were made of different sequences but have high similarity scores to two 20-bp motifs, were found from 11 human, 8 animal, 9 plant and 10 environmental samples, indicating that these two 20-bp motifs were broadly present in diverse S. epidermidis isolates. Thirty-one PCR-amplified reads of 16S rRNA genes, which were currently not in the dataset, were utilized to verify the feasibility of using two 20-bp motifs for ribotyping S. epidermidis sub-species. S. epidermidis S1, S3, but not S2, isolates on the human scalp carried a 20-bp sequence domain with high similarities to a 20-bp motif in the sequence pLogo. The phylogenetic tree showed that S. epidermidis S1, S2 and S3 were not from a single common ancestor. Two newly identified 20-bp motifs here, thus, provided reference nucleotide residues for ribotyping S. epidermidis.

Evaluating Xanthomonas oryzae pv. oryzae (Xoo) infection dynamics in rice for distribution routes and environmental reservoirs by molecular approaches

Xanthomonas oryzae pv. oryzae (Xoo) is a bacterial pathogen responsible for bacterial leaf blight (BLB) in rice, which can result in significant yield losses of up to 70%. A study evaluated the spread of Xoo in rice fields using environmental samples and employed colorimetric loop-mediated amplification (cLAMP) and PCR for detection. Quantitative PCR (qPCR) was used to measure infection levels. The research compared infection severity between a susceptible rice variety, Phitsanulok 2 (PSL2), and a resistant variety, PSL2-Xa21. Results showed that Xoo infection decreased from the leaves to the roots, but the bacteria persisted in soil and water for up to 12 and 6 weeks, respectively. The cLAMP assay, with the LpXoo4009 primer, effectively detected Xoo at low concentrations in both soil and water. Additionally, common grasses found in rice fields, such as Eriochloa procera, Echinochloa crus-galli and Chloris barbata were identified as temporary reservoirs for Xoo, facilitating its spread. The Xoo pathogen is distributed from infected leaves to roots and then from roots to the soil and nearby water. Grasses in the fields contribute to the perpetuation of the infection cycle serving as potential reservoirs that maintain the pathogen’s presence in the environment.

Probing Functional Thin Films with Grazing Incidence X-Ray Scattering: The Power of Indexing

Grazing incidence small- and wide-angle X-ray scattering (GISAXS, GIWAXS) has been widely applied for the study of functional thin films, be it for the characterization of nanostructured morphologies in block copolymers, nanocomposites, and nanoparticle assemblies, or for the packing and orientation of aromatic molecules or conjugated polymers. Solution-processed thin films are typically uniaxial powders, with a specific crystallographic plane oriented parallel to the substrate surface while ordered domains assume random orientations laterally. The convenient GISAXS/GIWAXS scattering geometry facilitates obtaining complete information about thin film structure as well as the ability to study samples in well-defined sample environments, as controlled by temperature, exposure to solvent vapor and drying, or coating processes. Moreover, with suitable X-ray sources and detectors, information about the ordering kinetics and phase transitions can be obtained down to the millisecond scale. The scattering geometry and an interactive graphical tool to index such scattering patterns will be discussed here. Furthermore, it will be demonstrated that proper indexing of the X-ray scattering patterns can provide deep insight into thin film structure–property relationships and the kinetics of structure formation. Recent examples of nanostructures and molecular organization in thin films will be discussed, as well as self-assembly processes leading to such structures.

Influence of mesh selectivity on risk assessment of marine microplastics.

In this study, environmental microplastic samples (>30μm) were collected from surface seawater and the water column, characterized, and used to assess ecological risks. The influence of mesh selectivity on ecological risks was also evaluated through subsampling. Results show that surface microplastic concentrations (>30μm) range from 92 to 3306 pieces/m3 along Japan's southwest coast, with significant increases at Stas. 2 and 1. Subsurface vertical concentration near Okinawa ranges from 991 to 1992 pieces/m3, with denser, more toxic polymers more frequently observed in deeper waters, suggesting that polymer types may be sorted by marine structure. Risk assessments revealed very high risks near main islands and populated regions, while remote regions had lower risks. Further analysis revealed that ecological risk estimates are significantly influenced by mesh selectivity, with variations in particle size distribution and polymer type composition resulting in changes of up to 100-fold at the same location when different mesh sizes were used, suggesting that current framework is not ideal for risk assessment of microplastics. This study is the first to demonstrate that samplers with different mesh sizes can lead to substantial differences in risk assessments, even at the same location. These findings underscore the critical impact of mesh selectivity on ecological risk estimates and highlight the need for standardized sampling protocols in microplastic research.

Detection of vaccine-derived poliovirus type 2 from sewage samples and public health response, Poland, November to December 2024.

In October and December 2024, circulating vaccine-derived poliovirus type 2 (cVDPV2) was detected from two wastewater samples in Poland during routine environmental surveillance. The first isolate was characterised and matched previous cVDPV2 isolates detected in Spain in September, as well as in Germany, Finland, and the United Kingdom in November and December 2024. In response to the event, active surveillance for acute flaccid paralysis (AFP) has been strengthened, and the frequency of environmental sample collection has been increased.