Metagenomic and Antibiotic Resistance Analysis of the Gut Microbiota in Larus relictus and Anatidae Species Inhabiting the Honghaizi Wetland of Ordos, Inner Mongolia, from 2021 to 2023.

Gut microbes thrive by utilising host energy and, in return, provide valuable benefits, akin to a symbiotic relationship. Here, metagenomic sequencing was performed to characterise and compare the community composition, diversity and antibiotic resistance of the gut microbiota of Relict gull (Larus relictus) and Anatidae species. Alpha diversity analysis revealed that the intestinal microbial richness of L. relictus was significantly lower than that of Anatidae, with distinct differences observed in microbial composition. Notably, the intestines of L. relictus harboured more pathogenic bacteria such as clostridium, which may contribute to the decline in their population and endangered status. A total of 117 strains of Escherichia coli were isolated, with 90.60% exhibiting full susceptibility to 21 antibiotics, while 25.3% exhibited significant biofilm formation. Comprehensive Antibiotic Resistance Database data indicated that glycopeptide resistance genes were the most prevalent type carried by migratory birds, alongside quinolone, tetracycline and lincosamide resistance genes. The abundance of resistance genes carried by migratory birds decreased over time. This metagenomic analysis provides valuable insights into the intestinal microbial composition of these wild bird species, offering important guidance for their conservation efforts, particularly for L. relictus, and contributing to our understanding of pathogen spread and antibiotic-resistant bacteria.

A Wohlfahrtiimonas chitiniclastica with a novel type of blaVEB-1-carrying plasmid isolated from a zebra in China.

Background: Wohlfahrtiimonas chitiniclastica is an emerging fly-borne zoonotic pathogen, which causes infections in immunocompromised patients and some animals. Herein, we reported a W. chitiniclastica BM-Y from a dead zebra in China. Methods: The complete genome sequencing of BM-Y showed that this isolate carried one chromosome and one novel type of blaVEB-1-carrying plasmid. Detailed genetic dissection was applied to this plasmid to display the genetic environment of blaVEB-1. Results: Three novel insertion sequence (IS) elements, namely ISWoch1, ISWoch2, and ISWoch3, were found in this plasmid. aadB, aacA1, and gcuG were located downstream of blaVEB-1, composing a gene cassette array blaVEB-1-aadB-aacA1-gcuG bracketed by an intact ISWoch1 and a truncated one, which was named the blaVEB-1 region. The 5'-RACE experiments revealed that the transcription start site of the blaVEB-1 region was located in the intact ISWoch1 and this IS provided a strong promoter for the blaVEB-1 region. Conclusion: The spread of the blaVEB-1-carrying plasmid might enhance the ability of W. chitiniclastica to survive under drug selection pressure and aggravate the difficulty in treating infections caused by blaVEB-1-carrying W. chitiniclastica. To the best of our knowledge, this is the first report of the genetic characterization of a novel blaVEB-1-carrying plasmid with new ISs from W. chitiniclastica.

Comparative genetic, biochemical, and biophysical analyses of the four E. coli ABCF paralogs support distinct functions related to mRNA translation.

Multiple paralogous ABCF ATPases are encoded in most genomes, but the physiological functions remain unknown for most of them. We herein compare the four Escherichia coli K12 ABCFs - EttA, Uup, YbiT, and YheS - using assays previously employed to demonstrate EttA gates the first step of polypeptide elongation on the ribosome dependent on ATP/ADP ratio. A Δ uup knockout, like Δ ettA , exhibits strongly reduced fitness when growth is restarted from long-term stationary phase, but neither Δ ybiT nor Δ yheS exhibits this phenotype. All four proteins nonetheless functionally interact with ribosomes based on in vitro translation and single-molecule fluorescence resonance energy transfer experiments employing variants harboring glutamate-to-glutamine active-site mutations (EQ 2 ) that trap them in the ATP-bound conformation. These variants all strongly stabilize the same global conformational state of a ribosomal elongation complex harboring deacylated tRNA Val in the P site. However, EQ 2 -Uup uniquely exchanges on/off the ribosome on a second timescale, while EQ 2 -YheS-bound ribosomes uniquely sample alternative global conformations. At sub-micromolar concentrations, EQ 2 -EttA and EQ 2 -YbiT fully inhibit in vitro translation of an mRNA encoding luciferase, while EQ 2 -Uup and EQ 2 -YheS only partially inhibit it at ~10-fold higher concentrations. Moreover, tripeptide synthesis reactions are not inhibited by EQ 2 -Uup or EQ 2 -YheS, while EQ 2 -YbiT inhibits synthesis of both peptide bonds and EQ 2 -EttA specifically traps ribosomes after synthesis of the first peptide bond. These results support the four E. coli ABCF paralogs all having different activities on translating ribosomes, and they suggest that there remains a substantial amount of functionally uncharacterized "dark matter" involved in mRNA translation.

Vibrio parahaemolyticus from Migratory Birds in China Carries an Extra Copy of tRNA-Gly and Plasmid-Mediated Quinolone Resistance Gene qnrD.

Vibrio parahaemolyticus is a marine bacterium coming from estuarine environments, where the migratory birds can easily be colonized by V. parahaemolyticus. Migratory birds may be important reservoirs of V. parahaemolyticus by growth and re-entry into the environment. To further explore the spreading mechanism of V. parahaemolyticus among marine life, human beings, and migratory birds, we aimed to investigate the characteristics of the genetic diversity, antimicrobial resistance, virulence genes, and a potentially informative gene marker of V. parahaemolyticus isolated from migratory birds in China. This study recovered 124 (14.55%) V. parahaemolyticus isolates from 852 fecal and environmental (water) samples. All of the 124 strains were classified into 85 known sequence types (STs), of which ST-2738 was most frequently identified. Analysis of the population structure using whole-genome variation of the 124 isolates illustrated that they grouped into 27 different clonal groups (CGs) belonging to the previously defined geographical populations VppX and VppAsia. Even though these genomes have high diversity, an extra copy of tRNA-Gly was presented in all migratory bird-carried V. parahaemolyticus isolates, which could be used as a potentially informative marker of the V. parahaemolyticus strains derived from birds. Antibiotic sensitivity experiments revealed that 47 (37.10%) isolates were resistant to ampicillin. Five isolates harbored the plasmid-mediated quinolone resistance (PMQR) gene qnrD, which has not previously been identified in this species. The investigation of antibiotic resistance provides the basic knowledge to further evaluate the risk of enrichment and reintroduction of pathogenic V. parahaemolyticus strains in migratory birds. IMPORTANCE The presence of V. parahaemolyticus in migratory birds' fecal samples implies that the human pathogenic V. parahaemolyticus strains may also potentially infect birds and thus pose a risk for zoonotic infection and food safety associated with re-entry into the environment. Our study firstly highlights the extra copy of tRNA as a potentially informative marker for identifying the bird-carried V. parahaemolyticus strains. Also, we firstly identify the plasmid-mediated quinolone resistance (PMQR) gene qnrD in V. parahaemolyticus. To further evaluate the risk of enrichment and reintroduction of pathogenic strains carried by migratory birds, we suggest conducting estuarine environmental surveillance to monitor the antibiotic resistance and virulence factors of bird-carried V. parahaemolyticus isolates.

Characteristics of Carbapenem-resistant Klebsiella pneumoniae in sewage from a tertiary hospital in Jilin Province, China.

Carbapenem-resistant Klebsiella pneumoniae (CRKP) infection is a serious problem in hospitals worldwide. We monitored a tertiary hospital in Changchun, Jilin Province, China, and found that CRKP was the major species among the carbapenem-resistant isolates in sewage. Subsequently, we evaluated the drug susceptibility, resistance genes, virulence genes, outer pore membrane protein-related genes (OmpK35 & OmpK 36), multi-locus sequence typing and replicons, biofilm formation capabilities, and resistance to chlorine-containing disinfectants among KP isolates. Identification of drug sensitivity, multiple resistance profiles were observed including 77 (82.80%) multidrug resistant (MDR), 16 (17.20%) extensive drug resistant (XDR). Some antibiotic resistance genes were detected, the most prevalent carbapenemase gene was blaKPC, and 16 resistance genes were associated with other antibiotics. In addition, 3 (3.23%) CRKP isolates demonstrated loss of OmpK-35 and 2 (2.15%) demonstrated loss of OmpK-36. In the detection of multi-locus sequence typing (MLST), 11 ST11 isolates carried virulence genes. The most common replicon type was IncFII. Biofilm-forming capabilities were demonstrated by 68.8% of the isolates, all of which were resistant to chlorine-containing disinfectants. The results of the study showed that antibiotic-resistant isolates, especially CRKP, could resist disinfectants in hospital wastewater, and improper treatment of hospital wastewater may lead to the spread of drug-resistant bacteria and their genes. Thus, these bacteria must be eliminated before being discharged into the municipal sewage system.

Automated Sleep Staging via Parallel Frequency-Cut Attention.

Stage-based sleep screening is a widely-used tool in both healthcare and neuroscientific research, as it allows for the accurate assessment of sleep patterns and stages. In this paper, we propose a novel framework that is based on authoritative guidance in sleep medicine and is designed to automatically capture the time-frequency characteristics of sleep electroencephalogram (EEG) signals in order to make staging decisions. Our framework consists of two main phases: a feature extraction process that partitions the input EEG spectrograms into a sequence of time-frequency patches, and a staging phase that searches for correlations between the extracted features and the defining characteristics of sleep stages. To model the staging phase, we utilize a Transformer model with an attention-based module, which allows for the extraction of global contextual relevance among time-frequency patches and the use of this relevance for staging decisions. The proposed method is validated on the large-scale Sleep Heart Health Study dataset and achieves new state-of-the-art results for the wake, N2, and N3 stages, with respective F1 scores of 0.93, 0.88, and 0.87 using only EEG signals. Our method also demonstrates high inter-rater reliability, with a kappa score of 0.80. Moreover, we provide visualizations of the correspondence between sleep staging decisions and features extracted by our method, which enhances the interpretability of the proposal. Overall, our work represents a significant contribution to the field of automated sleep staging and has important implications for both healthcare and neuroscience research.

Learning vector quantized representation for cancer subtypes identification.

BACKGROUND AND OBJECTIVE: Defining and separating cancer subtypes is essential for facilitating personalized therapy modality and prognosis of patients. The definition of subtypes has been constantly recalibrated as a result of our deepened understanding. During this recalibration, researchers often rely on clustering of cancer data to provide an intuitive visual reference that could reveal the intrinsic characteristics of subtypes. The data being clustered are often omics data such as transcriptomics that have strong correlations to the underlying biological mechanism. However, while existing studies have shown promising results, they suffer from issues associated with omics data: sample scarcity and high dimensionality while they impose unrealistic assumptions to extract useful features from the data while avoiding overfitting to spurious correlations. METHODS: This paper proposes to leverage a recent strong generative model, Vector-Quantized Variational AutoEncoder, to tackle the data issues and extract discrete representations that are crucial to the quality of subsequent clustering by retaining only information relevant to reconstructing the input. RESULTS: Extensive experiments and medical analysis on multiple datasets comprising 10 distinct cancers demonstrate the proposed clustering results can significantly and robustly improve prognosis over prevalent subtyping systems. CONCLUSION: Our proposal does not impose strict assumptions on data distribution; while, its latent features are better representations of the transcriptomic data in different cancer subtypes, capable of yielding superior clustering performance with any mainstream clustering method.

Dietary nano-Se supplementation regulates lipid deposition, protein synthesis and muscle fibre formation in grass carp fed with high-fat diet.

The current study aims to confirm the positive effects of dietary nano-Se on nutrients deposition and muscle fibre formation in grass carp fed with high-fat diet (HFD) before overwintering and to reveal its possible molecular mechanism. The lipid deposition, protein synthesis and muscle fibre formation in grass carp fed with regular diet (RD), HFD or HFD supplemented with nano-Se (0·3 or 0·6 mg/kg) for 60 d were tested. Results show that nano-Se significantly reduced lipid content, dripping loss and fibre diameter (P < 0·05), but increased protein content, post-mortem pH24 h and muscle fibre density (P < 0·05) in muscle of grass carp fed with HFD. Notably, dietary nano-Se decreased lipid deposition in the muscle by regulating amp-activated protein kinase activity and increased protein synthesis and fibre formation in muscle by activating target of rapamycin and myogenic determining factors pathways. In summary, dietary nano-Se can regulate the nutrients deposition and muscle fibre formation in grass carp fed with HFD, which exhibit potential benefit for improving flesh quality of grass carp fed with HFD.

Virulence genes and antimicrobial resistance in Enterococcus strains isolated from dogs and cats in Northeast China.

This study aimed to characterize the antimicrobial resistance and virulence of Enterococcus from dogs and cats in Northeast China and evaluate its zoonotic risk based on a total of 469 enterococci strains from 610 samples, including 238 strains of E. faecium and 128 strains of E. faecalis. The isolation rate from police dog samples was 93.79%, pet dog samples was 69.90% and pet cat samples was 76.67%. The differences in the prevalence of E. faecalis among different hosts were statistically significant (P<0.05). The assays showed that most of the virulence genes detected were existed in E. faecalis and police dogs carried the least number of virulence genes. The correlation between enterococcal surface protein (esp) and aggregation substance (asa1) was determined. Enterococci are most resistant to tetracycline and erythromycin, 68.92% of the isolates were classified as multiple drug resistant. Significant differences (P<0.01) were found between E. faecium and E. faecalis in the resistance rates of nine antimicrobials. Four positive and four negative correlations were found between virulence genes and antimicrobial resistance. The results show that Enterococcus colonization and excretion in dogs and cats were related to animal species and living environments. Some correlation between virulence factors and antimicrobial resistance was obtained. This study confirmed the presence of strains carrying multiple virulence factors and antimicrobial resistance at the same time, suggesting a public health risk for dogs and cats as reservoirs of enterococci.

Complete genome sequence of a blaNDM-5-producing Escherichia coli DC71 assigned as ST410-O8:H9 and recovered from a captive giant panda (Ailuropoda melanoleuca) in China.

OBJECTIVES: In this study, we report the complete genome sequence of a multidrug-resistant Escherichia coli strain recovered from a fecal sample from a captive giant panda in China. METHODS: Antimicrobial susceptibility testing was performed. Genomic DNA from E. coli DC71 was sequenced using a Nanopore PromethION sequencer instrument (Oxford Nanopore Technologies, UK) and MGI High-throughput Sequencing MGISEQ-2000 platforms. The clean reads were de novo assembled using SPAdes v3.11. The complete genome was annotated and analyzed using multilocus sequence typing, serotyping, plasmid replicons, fimH typing, chromosomal point mutations, acquired antimicrobial resistance, and virulence genes with web tools available at the Center for Genomic Epidemiology. RESULTS: The complete genome, 4 991 906 bp in length and comprising 4677 protein-coding sequences, was generated. In silico analysis revealed that E. coli DC71 belonged to the ST410-O8:H9 subclone. A carbapenem resistance gene, blaNDM-5, was located on the pDC71-2 plasmid, coproducing blaTEM-1. Many other resistance determinants encoded by chromosomes and pDC71-3 were found. The virulence related genes carried by chromosomes were mostly related to enterohemorrhagic E. coli (EHEC) O157:H7. CONCLUSIONS: To our knowledge, this is the first complete genome of an E. coli ST410-O8:H9 strain recovered from captive giant panda in China. This multidrug-resistant E. coli subclone may pose potential risks to human and animal health. The genome sequence will be helpful to understand the genomic structure, its diversity, and the molecular mechanism allowing bacteria to disseminate the resistance gene.

Case Report: Transvertebral transposition of the spinal cord for recovery after paraplegia during kyphoscoliosis surgery.

Introduction: Neurological impairment during spinal deformity surgery is the most serious possible complication. When confronting intraoperative neurophysiological monitoring alerts, various surgical management methods such as the release of implants and decompression of the spinal cord are always performed. Transvertebral transposition of the spinal cord is rarely performed, and its role in the management of acute paraplegia is seldom reported. Case description: The authors present two patients with kyphoscoliosis who experienced neurological deficits and abnormal neurological monitoring intraoperatively or post-operatively that were detected during correction surgery. Acute paraplegia was confirmed by a wake-up test. Subsequent spinal cord transposition was performed. Intraoperative neurophysiological monitoring motor-evoked potentials (MEPs) and somatosensory-evoked potentials (SEPs) were performed to detect the changes during the process. After transvertebral transposition of the spinal cord, the MEPs and SEPs were significantly improved in both patients during surgery. The spinal cord function was restored post-operatively and recovered to normal at the final follow-up in two patients. Conclusion: This case demonstrated that instead of decreasing the correction ratio of kyphoscoliosis, transvertebral transposition of the spinal cord under intraoperative neurophysiological monitoring may be an alternative therapeutic strategy for acute spinal cord dysfunction caused by deformity correction surgeries.

Antimicrobial Resistance and Prevalence of Extended Spectrum ß-Lactamase-Producing Escherichia coli from Dogs and Cats in Northeastern China from 2012 to 2021.

(1) Background: there has been a growing concern about pet-spread bacterial zoonosis in recent years. This study aimed to investigate the trend in drug-resistance of canine Escherichia coli isolates in northeast China between 2012-2021 and the differences in drug-resistance of E. coli of different origins in 2021. (2) Methods: E. coli were isolated from feces or anal swab samples from dogs and cats, and their antibiotic susceptibility profiles and phylogenetic grouping were identified. PCR was applied on the extended spectrum ß-lactamase (ESBL) E. coli for antibiotic resistance genes. (3) Results: five hundred and fifty-four E. coli isolates were detected in 869 samples (63.75%). The multidrug resistance (MDR) rates of E. coli in pet dogs showed a decreasing trend, but working dogs showed the opposite trend. Resistance genes blaCTX-M and blaCTX-M+TEM were dominant among the ESBL producers (n = 219). The consistency between the resistance phenotypes and genes was high except for fluoroquinolone-resistant ESBL E. coli. All ESBL E. coli-carrying blaNDM were isolated from working dogs, and one of the strains carried mcr-1 and blaNDM-4. Phylogroup B2 was the dominant group in pet cats, and more than half of the isolates from companion cats were ESBL E. coli. (4) Conclusions: the measures taken to reduce resistance in China were beginning to bear fruit. Companion cats may be more susceptible to colonization by ESBL E. coli. The problem of resistant bacteria in working dogs and pet cats warrants concern.

Cancer Subtyping via Embedded Unsupervised Learning on Transcriptomics Data.

Cancer is one of the deadliest diseases worldwide. Accurate diagnosis and classification of cancer subtypes are indispensable for effective clinical treatment. Promising results on automatic cancer subtyping systems have been published recently with the emergence of various deep learning methods. However, such automatic systems often overfit the data due to the high dimensionality and scarcity. In this paper, we propose to investigate automatic subtyping from an unsupervised learning perspective by directly constructing the underlying data distribution itself, hence sufficient data can be generated to alleviate the issue of overfitting. Specifically, we bypass the strong Gaussianity assumption that typically exists but fails in the unsupervised learning subtyping literature due to small-sized samples by vector quantization. Our proposed method better captures the latent space features and models the cancer subtype manifestation on a molecular basis, as demonstrated by the extensive experimental results.

Adaptive Spike-Like Representation of EEG Signals for Sleep Stages Scoring.

Recently there has seen promising results on auto-matic stage scoring by extracting spatio-temporal features from electroencephalogram (EEG). Such methods entail laborious manual feature engineering and domain knowledge. In this study, we propose an adaptive scheme to probabilistically encode, filter and accumulate the input signals and weight the resultant features by the half-Gaussian probabilities of signal intensities. The adaptive representations are subsequently fed into a transformer model to automatically mine the relevance between features and corresponding stages. Extensive exper-iments on the largest public dataset against state-of-the-art methods validate the effectiveness of our proposed method and reveal promising future directions.

A clinical Pseudomonas juntendi strain with bla IMP-1 carried by an integrative and conjugative element in China.

Objective: To precisely determine the species of a carbapenem-resistant Pseudomonas strain 1809276 isolated from the urine of a Chinese patient and analyze its integrative and conjugative element (ICE) 1276 formation mechanism. Methods: Single-molecule real-time (SMRT) sequencing was carried out on strain 18091276 to obtain the complete chromosome and plasmid (pCN1276) sequences, and average nucleotide identity (ANI) was used for precise species identification. The ICEs in GenBank with the same integrase structure as ICE 1276 were aligned. At the same time, the transfer ability of bla IMP-1 and the antibiotic sensitivity of Pseudomonas juntendi 18091276 were tested. Results: This bacterium was P. juntendi, and its drug resistance mechanism is the capture of the accA4' gene cassette by the Tn402-like type 1 integron (IntI1-bla IMP-1) to form In1886 before its capture by the ΔTn4662a-carrying ICE 1276. The acquisition of bla IMP-1 confers carbapenem resistance to P. juntendi 18091276. Conclusion: The formation of bla IMP-1-carrying ICE 1276, its further integration into the chromosomes, and transposition and recombination of other elements promote bacterial gene accumulation and transmission.

First Report of the Colistin Resistance Gene mcr-10.1 Carried by IncpA1763-KPC Plasmid pSL12517-mcr10.1 in Enterobacter cloacae in Sierra Leone.

Mobile colistin resistance (mcr) gene mcr-10.1 has been distributed widely since it was initially identified in 2020. The aim of this study was to report the first mcr-10.1 in Africa and the first mcr in Sierra Leone; furthermore, we presented diverse modular structures of mcr-10.1 loci. Here, the complete sequence of one mcr-10.1-carrying plasmid in one clinical Enterobacter cloacae isolate from Sierra Leone was determined. Detailed genetic dissection and comparison were applied to this plasmid, together with a homologous plasmid carrying mcr-10.1 from GenBank. Moreover, a genetic comparison of 19 mcr-10.1 loci was performed. In this study, mcr-10.1 was carried by an IncpA1763-KPC plasmid from one Enterobacter cloacae isolate. A total of 19 mcr-10.1 loci displayed diversification in modular structures through complex transposition and homologous recombination. A site-specific tyrosine recombinase XerC was located upstream of mcr-10.1, and at least one insertion sequence element was inserted adjacent to a conserved xerC-mcr-10.1-orf336-orf177 region. Integration of mcr-10.1 into a different gene context and carried by various Inc plasmids contributed to the wide distribution of mcr-10.1 and enhanced the ability of bacteria to survive under colistin selection pressure. IMPORTANCE Colistin is used as one of the last available choices of antibiotics for patients infected by carbapenem-resistant bacterial strains, but the unrestricted use of colistin aggravated the acquisition and dissemination of mobile colistin resistance (mcr) genes. So far, 10 mcr genes have been reported in four continents around the world. This study presented one mcr-10.1-carrying Enterobacter cloacae isolate from Sierra Leone. The mcr-10.1 gene was identified on an IncpA1763-KPC plasmid. According to the results of genetic comparison of 19 mcr-10.1 loci, the mcr-10.1 gene was found to be located in a conserved xerC-mcr-10.1-orf336-orf177 region, and at least one insertion sequence element was inserted adjacent to this region. To our knowledge, this is the first report of identifying the mcr-10.1 gene in Africa and the mcr gene in Sierra Leone.

Molecular Characteristics of Extended-Spectrum Beta-Lactamase-Producing Escherichia coli Strains Isolated from Diseased Captive Giant Pandas (Ailuropoda melanoleuca) in China.

Objectives: To characterize the antimicrobial resistance and virulence of pathogenic Escherichia coli isolated from diseased captive giant pandas. Methods: Antimicrobial susceptibility and minimum inhibitory concentration (MIC) were determined by the broth dilution method. Whole-genome sequencing was used to characterize the phylogeny, serotype, virulence, resistome, plasmids, and genetic structures of the cefotaxime (CTX)-M genes. Results: Four extended-spectrum beta-lactamase (ESBL)-producing E. coli strains were identified and the MICs against 11 antibiotics in vitro were determined. All ESBL-producing E. coli strains were resistant to more than eight antibiotics and carried the blaCTX-M-55 or blaCTX-M-105 gene in different sizes of replicon-type plasmids (pAMSH1-IncHI2, 257 kb; pAMPD2-IncFII, 89 kb; pAMPD02-IncFIB, 129 kb; and pAMSC4-IncN, 47 kb). Distinct insertional sequences and transposases were identified up-/downstream of blaCTX-Ms, including IS26, ISEcp1, ISKpn72, IS903B, and Tn2. These strains also possessed at least three virulence genes of pathogenic E. coli and originated in four different evolutionary branches. One strain carried the complete locus of the enterocyte effacement pathogenicity island, but lacked the virulence genes stx and bfpA, indicating atypical enteropathogenic E. coli, whereas the other strains were considered to be extraintestinal pathogenic E. coli. Conclusions: The emergence of ESBL-producing pathogenic E. coli strains from diseased captive giant pandas warrants greater attention. The findings of this study will help to prevent the spread of these strains among captive giant pandas as well as from wild animals to humans.

Detection of Two Copies of a bla NDM-1-Encoding Plasmid in Escherichia coli Isolates from a Pediatric Patient with Diarrhea.

PURPOSE: To elucidate the contribution of a transferable plasmid harboring the bla NDM-1 gene in an Escherichia coli clinical isolate to the spread of resistance determinants. METHODS: Nine extended-spectrum ß-lactamase-producing E. coli were collected from diarrhea samples from a pediatric patient and genetic linkage was investigated through enterobacteriaceae repetitive intragenic consensus polymerase chain reaction (PCR). Bacterial species were identified by 16s rRNA sequencing, susceptibility testing with the use of a BD PhoenixTM-100 Automated Microbiology System, and assessment of virulence genes by PCR. The transferability of bla NDM-1 in E. coli strain TCM3e1 was confirmed by conjugation experiments. Complete sequencing of E. coli strain TCM3e1 was determined with the PacBio and Illumina NovaSeq platforms and the characteristics were analyzed with bioinformatics software. RESULTS: The results showed that all nine E. coli strains were the same clone. E. coli strain TCM3e1 was resistant to 12 antimicrobial agents and carried the virulence gene EAST-1. Conjugation transfer analysis showed that bla NDM-1 was carried on a self-transmissible plasmid. Two copies of the bla NDM-1 gene were present on an IncC plasmid and some resistance genes with two or three copies were located downstream of the bla NDM-1 gene and formed a tandem repeat fragment (bla DNM-1-bleo-sul1- aadA17- dfrA12). CONCLUSION: A transmissible plasmid harboring two copies of the bla NDM-1 gene, including clonal dispersions of the bla NDM-1 gene, was identified in clinical isolates. These findings emphasized the necessity of surveillance of the plasmid-borne bla NDM-1 to prevent dissemination.

Microplastics removal and characteristics of constructed wetlands WWTPs in rural area of Changsha, China: A different situation from urban WWTPs.

Wastewater treatment plants (WWTPs) are important pathways that discharged microplastics into the natural environment, but few relevant research has been conducted in rural areas, especially with horizontal subsurface flow constructed wetlands (HSSFCWs). This study systematically investigates the removal efficiency and characteristics of microplastics in two rural WWTPs with HSSFCW in Changsha city of China and compared the microplastic pollution data of urban and rural WWTPs, to provide some advice for improving the microplastics removal efficiencies in rural WWTPs. 3 L wastewater were collected at each sampling point. Then microplastics in wastewater were extracted by density separation. The size, shape, color, and type of microplastics were analyzed and identified using the integrated microscope and FTIR. The whole experiment was carried out about a month. The results showed that the microplastics removal efficiency of rural WWTP1 was 72.38%, and that of rural WWTP2 was 68.10%, which were lower than that of most urban WWTPs. The microplastics removal efficiency of constructed wetlands in rural WWTP1 was 26.59%, and that in rural WWTP2 was 10.61%. Based on the daily discharge volume and the abundance of microplastics in the effluent of WWTPs, approximately 1.45 ∗ 107 items and 1.73 ∗ 107 items of microplastics were released each day from two rural WWTPs, separately. Fiber was the primary microplastic in both influent and effluent. The polyethylene (PE) and polystyrene (PS) were the main ingredients. The primary source of microplastics in rural WWTPs was inferred as domestic sewage. Microplastics removal efficiencies of rural WWTPs can be improved by regular maintenance, reducing the grid spacing, increasing the hydraulic stay time of biochemical pool, and increasing plant density, changing plant species, or adjusting the size and fill order of matrix in HSSFCWs, which can effectively help to prevent secondary pollution of microplastics from rural WWTPs.

Virulence, Antibiotic Resistance, and Phylogenetic Relationships of Aeromonas spp. Carried by Migratory Birds in China.

This study aimed to evaluate antimicrobial resistance, virulence, and the genetic diversity of Aeromonas isolated from migratory birds from Guangxi Province, Guangdong Province, Ningxia Hui Autonomous Region, Jiangxi Province, and Inner Mongolia in China. A total of 810 samples were collected, including fresh feces, cloacal swabs, and throat swabs. The collected samples were processed and subjected to bacteriological examination. The resistance to 21 antibiotics was evaluated. A phylogenetic tree was constructed using concatenated gltA-groL-gyrB-metG-PPSA-recA sequences. Eight putative virulence factors were identified by PCR and sequencing, and a biofilm formation assay was performed using a modified microtiter plate method. In total, 176 Aeromonas isolates were isolated including A. sobria, A. hydrophila, A. veronii, and A. caviae. All isolates showed variable resistance against all 16 tested antibiotic discs, and only one antibiotic had no reference standard. Six kinds of virulence gene markers were discovered, and the detection rates were 46.0% (hlyA), 76.1% (aerA), 52.3% (alt), 4.5% (ast), 54.0% (fla), and 64.2% (lip). These strains were able to form biofilms with distinct magnitudes; 102 were weakly adherent, 14 were moderately adherent, 60 were non-adherent, and none were strongly adherent. Our results suggest that migratory birds carry highly virulent and multidrug-resistant Aeromonas and spread them around the world through migration, which is a potential threat to public health.