Genetic Analyses of Rare ESBL ST628 Klebsiella pneumoniae Detected during a Protracted Nosocomial Outbreak in the United Kingdom.

Klebsiella pneumoniae (K. pneumoniae) cultures from a hospital-wide outbreak in the UK, which lasted for over 12 months, were sequenced. We sought to sequence and genetically characterise the outbreak strain. Antibiotic Susceptibility Testing (AST) was performed on 65 K. pneumoniae isolates saved from the outbreak. All isolates were sequenced using the Oxford Nanopore Technologies (ONT) MinION flowcell: 10 isolates, including the isolate with the earliest collection date in 2017, were additionally sequenced on the NovaSeq 6000 platform to build high-accuracy nanopore-illumina assemblies. Among the sequenced strains, 60 were typed as ST628. 96.6% (n = 58/60) ST628 strains harboured a large ~247-kb FIB(K) plasmid carrying up to 11 antimicrobial resistance genes, including the extended-spectrum beta-lactamase (ESBL) gene, blaCTX-M-15. Clonality between the outbreak isolates was confirmed using single nucleotide polymorphism (SNP) typing. The outbreak strains were phylogenetically related to clinical ST628 strains identified in 2012, 6 years prior to the outbreak. A rare ESBL K. pneumoniae K2 ST628 strain harbouring a multi-drug resistant (MDR) plasmid encoding the ESBL gene blaCTX-M-15 was detected across multiple independent wards during the protracted nosocomial outbreak. Surveillance of this strain is recommended to prevent future nosocomial outbreaks.

Distinct microplastic patterns in the sediment and biota of an urban stream.

Urban freshwaters, their sediments and resident biota are often highly susceptible to microplastic contamination from catchment-specific sources. Water velocity and spatiotemporal dynamics within the system can impact microplastic loads, while biological features may additionally impact levels within freshwater biota. Here, we investigated the spatiotemporal variations in microplastic loads collected from sediment, macroinvertebrate and fish samples from an urban watercourse (Bourne Stream) in Dorset, southwest England. Sediment particles were mostly fragments of colours (especially orange and purple) whereas microplastics in both macroinvertebrates and fishes were blue/green and fibres. Across all sample types, the dominant particle size class was ≤100 µm. Median (M) and range (R) of microplastic loads within each sample type were sediment: M = 0.06, R = 0-0.36 particles g-1; macroinvertebrates: M = 0, R = 0-4 particles per batch; and fishes: M = 1, R = 0-6 particles per individual. Sediment loads varied spatially, with the highest load in the most upstream site, whereas biotic loads did not vary across space and time. Macroinvertebrate batch loadings varied between taxa and feeding guild, with counts significantly higher in annelids but lower in herbivores. Fish counts were higher in species with true, differentiated stomachs, but with the effects of species, feeding guild and body size being non-significant. Within sites, mean microplastic loads did not correlate between sediment, macroinvertebrate and fish samples. These results suggest that sediment freshwater microplastic loadings may vary spatially but that these trends are not reflected by, or correlated to, those in the biota where ingestion varies with biological traits. Assessments of freshwater microplastic contamination must therefore consider sampling spatiotemporally and across different biotic communities to fully understand the scale of contamination, and to subsequently undertake effective mitigation steps.

Microplastic loads within riverine fishes and macroinvertebrates are not predictable from ecological or morphological characteristics.

Microplastics are a relatively new but important form of freshwater contamination that can be ingested by a range of different species, with particle counts thought to be predictable from species ecology and morphology. Here, we report levels of microplastics in a 26 µm-5 mm size range within the macroinvertebrate and fish community of a lowland river (Dorset Stour, SW England), and test the hypothesis that counts are predictable from characteristics such as feeding guild, body length and trophic position. Macroinvertebrates (n = 257, 12 taxa) and fish (n = 418, 9 species) were collected from distinct river reaches by kick sampling and rod and line angling, respectively. Batches of whole macroinvertebrates and individual fish gastrointestinal tracts were digested with 30% hydrogen peroxide before microplastic screening and FTIR polymer confirmation on a particle subset. Particles were found in 40% of pooled macroinvertebrate batches (taxa incidences: 14-75%) and 39% of fishes (species incidences: 29-47%). Dominant particle feature categories were ≤100 µm, blue/green, fragments and fibres identified as various polyolefins. Although particle counts in macroinvertebrates were highest in Ephemeroptera (mean of 0.74 particles per individual), the relationships between particle loads, batch number and guild were all non-significant. In fishes, particle counts were not significantly related to species, stomach structure, feeding guild or body length, with spatial differences also not apparent across the catchment. Individual fish particle counts were similarly not significantly associated with their trophic positions (calculated from bulk δ15N values for a subset of fishes) and parasite load of Pomphorhynchus tereticollis. Correlations between fish and macroinvertebrate particle counts within specific river reaches were also not significant. In entirety, these results indicated although loadings of microplastic particles were relatively consistent within the two communities, they were not predictable from any of their ecological or morphological characteristics.