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1.
BMC Microbiol ; 24(1): 225, 2024 Jun 26.
Article in English | MEDLINE | ID: mdl-38926687

ABSTRACT

BACKGROUND: The incidence of hospital-acquired infections in extensively drug-resistant Pseudomonas aeruginosa (XDR-PA) has been increasing worldwide and is frequently associated with an increase in mortality and morbidity rates. The aim of this study was to characterize clinical XDR-PA isolates recovered during six months at three different hospitals in Egypt. RESULTS: Seventy hospital-acquired clinical isolates of P. aeruginosa were classified into multidrug-resistant (MDR), extensively drug-resistant (XDR) and pandrug-resistant (PDR), according to their antimicrobial resistance profile. In addition, the possession of genes associated with mobile genetic elements and genes encoding antimicrobial resistance determinants among isolates were detected using polymerase chain reaction. As a result, a significant percentage of the isolates (75.7%) were XDR, while 18.5% were MDR, however only 5.7% of the isolates were non-MDR. The phenotypic detection of carbapenemases, extended-spectrum ß-lactamases (ESBLs) and metallo ß-lactamase (MBL) enzymes showed that 73.6% of XDR-PA isolates were carbapenemases producers, whereas 75.5% and 88.7% of XDR-PA isolates produced ESBLs and MBL respectively. In addition, PCR screening showed that oxa gene was the most frequently detected gene of carbapenemases (91.4%), while aac(6')-lb gene was mostly detected (84.3%) among the screened aminoglycosides-resistance genes. Furthermore, the molecular detection of the colistin resistance gene showed that 12.9% of isolates harbored mcr-1 gene. Concerning mobile genetic element markers (intI, traA, tnp513, and merA), intI was the highest detected gene as it was amplified in 67 isolates (95.7%). Finally, phylogenetic and molecular typing of the isolates via ERIC-PCR analysis revealed 10 different ERIC fingerprints. CONCLUSION: The present study revealed a high prevalence of XDR-PA in hospital settings which were resistant to a variety of antibiotics due to several mechanisms. In addition, 98% of the XDR-PA clinical isolates contained at least one gene associated with movable genetic elements, which could have aided the evolution of these XDR-PA strains. To reduce spread of drug resistance, judicious use of antimicrobial agents and strict infection control measures are therefore essential.


Subject(s)
Anti-Bacterial Agents , Cross Infection , Drug Resistance, Multiple, Bacterial , Microbial Sensitivity Tests , Pseudomonas Infections , Pseudomonas aeruginosa , beta-Lactamases , Pseudomonas aeruginosa/genetics , Pseudomonas aeruginosa/drug effects , Pseudomonas aeruginosa/isolation & purification , Humans , Pseudomonas Infections/microbiology , Pseudomonas Infections/epidemiology , Drug Resistance, Multiple, Bacterial/genetics , Cross Infection/microbiology , Cross Infection/epidemiology , Egypt/epidemiology , beta-Lactamases/genetics , Anti-Bacterial Agents/pharmacology , Bacterial Proteins/genetics , Hospitals/statistics & numerical data , Interspersed Repetitive Sequences/genetics , Polymerase Chain Reaction
2.
Sci Rep ; 14(1): 13056, 2024 06 06.
Article in English | MEDLINE | ID: mdl-38844487

ABSTRACT

Metagenomics has made it feasible to elucidate the intricacies of the ruminal microbiome and its role in the differentiation of animal production phenotypes of significance. The search for mobile genetic elements (MGEs) has taken on great importance, as they play a critical role in the transfer of genetic material between organisms. Furthermore, these elements serve a dual purpose by controlling populations through lytic bacteriophages, thereby maintaining ecological equilibrium and driving the evolutionary progress of host microorganisms. In this study, we aimed to identify the association between ruminal bacteria and their MGEs in Nellore cattle using physical chromosomal links through the Hi-C method. Shotgun metagenomic sequencing and the proximity ligation method ProxiMeta were used to analyze DNA, getting 1,713,111,307 bp, which gave rise to 107 metagenome-assembled genomes from rumen samples of four Nellore cows maintained on pasture. Taxonomic analysis revealed that most of the bacterial genomes belonged to the families Lachnospiraceae, Bacteroidaceae, Ruminococcaceae, Saccharofermentanaceae, and Treponemataceae and mostly encoded pathways for central carbon and other carbohydrate metabolisms. A total of 31 associations between host bacteria and MGE were identified, including 17 links to viruses and 14 links to plasmids. Additionally, we found 12 antibiotic resistance genes. To our knowledge, this is the first study in Brazilian cattle that connect MGEs with their microbial hosts. It identifies MGEs present in the rumen of pasture-raised Nellore cattle, offering insights that could advance biotechnology for food digestion and improve ruminant performance in production systems.


Subject(s)
Interspersed Repetitive Sequences , Rumen , Animals , Cattle , Rumen/microbiology , Interspersed Repetitive Sequences/genetics , Metagenomics/methods , Metagenome , Microbiota/genetics , Gastrointestinal Microbiome/genetics , Bacteria/genetics , Bacteria/classification , Genome, Bacterial , Phylogeny
3.
Methods Mol Biol ; 2813: 19-37, 2024.
Article in English | MEDLINE | ID: mdl-38888768

ABSTRACT

Genomics has revolutionized how we characterize and monitor infectious diseases for public health. The surveillance and characterization of Salmonella has improved drastically within the past decade. In this chapter, we discuss the prerequisites for good bacterial genomics studies and make note of advantages and disadvantages of this research approach. We discuss methods for outbreak detection and the evolutionary and epidemiological characterization of Salmonella spp. We provide an outline for determining the sequence type and serotype of isolates, building a core genome phylogenetic tree, and detecting antimicrobial resistance genes, virulence factors, and mobile genetic elements. These methods can be used to study other pathogenic bacterial species.


Subject(s)
Genome, Bacterial , Genomics , Molecular Epidemiology , Phylogeny , Salmonella Infections , Salmonella , Salmonella/genetics , Humans , Genomics/methods , Salmonella Infections/microbiology , Salmonella Infections/epidemiology , Molecular Epidemiology/methods , Virulence Factors/genetics , Disease Outbreaks , Drug Resistance, Bacterial/genetics , Interspersed Repetitive Sequences/genetics
4.
Microbiome ; 12(1): 107, 2024 Jun 14.
Article in English | MEDLINE | ID: mdl-38877573

ABSTRACT

BACKGROUND: Aquaculture is an important food source worldwide. The extensive use of antibiotics in intensive large-scale farms has resulted in resistance development. Non-intensive aquaculture is another aquatic feeding model that is conducive to ecological protection and closely related to the natural environment. However, the transmission of resistomes in non-intensive aquaculture has not been well characterized. Moreover, the influence of aquaculture resistomes on human health needs to be further understood. Here, metagenomic approach was employed to identify the mobility of aquaculture resistomes and estimate the potential risks to human health. RESULTS: The results demonstrated that antibiotic resistance genes (ARGs) were widely present in non-intensive aquaculture systems and the multidrug type was most abundant accounting for 34%. ARGs of non-intensive aquaculture environments were mainly shaped by microbial communities accounting for 51%. Seventy-seven genera and 36 mobile genetic elements (MGEs) were significantly associated with 23 ARG types (p < 0.05) according to network analysis. Six ARGs were defined as core ARGs (top 3% most abundant with occurrence frequency > 80%) which occupied 40% of ARG abundance in fish gut samples. Seventy-one ARG-carrying contigs were identified and 75% of them carried MGEs simultaneously. The qacEdelta1 and sul1 formed a stable combination and were detected simultaneously in aquaculture environments and humans. Additionally, 475 high-quality metagenomic-assembled genomes (MAGs) were recovered and 81 MAGs carried ARGs. The multidrug and bacitracin resistance genes were the most abundant ARG types carried by MAGs. Strikingly, Fusobacterium_A (opportunistic human pathogen) carrying ARGs and MGEs were identified in both the aquaculture system and human guts, which indicated the potential risks of ARG transfer. CONCLUSIONS: The mobility and pathogenicity of aquaculture resistomes were explored by a metagenomic approach. Given the observed co-occurrence of resistomes between the aquaculture environment and human, more stringent regulation of resistomes in non-intensive aquaculture systems may be required. Video Abstract.


Subject(s)
Anti-Bacterial Agents , Aquaculture , Metagenomics , Humans , Metagenomics/methods , Anti-Bacterial Agents/pharmacology , Animals , Bacteria/genetics , Bacteria/classification , Bacteria/drug effects , Bacteria/isolation & purification , Metagenome , Fishes/microbiology , Drug Resistance, Bacterial/genetics , Drug Resistance, Microbial/genetics , Genes, Bacterial/genetics , Interspersed Repetitive Sequences/genetics
5.
Cell Host Microbe ; 32(6): 804-819, 2024 Jun 12.
Article in English | MEDLINE | ID: mdl-38870898

ABSTRACT

Bacteriophages and other mobile genetic elements (MGEs) pose a significant threat to bacteria, subjecting them to constant attacks. In response, bacteria have evolved a sophisticated immune system that employs diverse defensive strategies and mechanisms. Remarkably, a growing body of evidence suggests that most of these defenses are encoded by MGEs themselves. This realization challenges our traditional understanding of bacterial immunity and raises intriguing questions about the evolutionary forces at play. Our review provides a comprehensive overview of the latest findings on the main families of MGEs and the defense systems they encode. We also highlight how a vast diversity of defense systems remains to be discovered and their mechanism of mobility understood. Altogether, the composition and distribution of defense systems in bacterial genomes only makes sense in the light of the ecological and evolutionary interactions of a complex network of MGEs.


Subject(s)
Bacteria , Bacteriophages , Interspersed Repetitive Sequences , Bacteria/genetics , Bacteria/immunology , Bacteriophages/genetics , Genome, Bacterial
6.
J Microorg Control ; 29(2): 55-65, 2024.
Article in English | MEDLINE | ID: mdl-38880617

ABSTRACT

Cupriavidus metallidurans strain PD11 isolated from laboratory waste drainage can use C1 compounds, such as dichloromethane (DCM) and methanol, as a sole carbon and energy source. However, strain CH34 (a type-strain) cannot grow in the medium supplemented with DCM. In the present study, we aimed to unravel the genetic elements underlying the utilization of C1 compounds by strain PD11. The genome subtraction approach indicated that only strain PD11 had several genes highly homologous to those of Herminiimonas arsenicoxydans strain ULPAs1. Moreover, a series of polymerase chain reaction (PCR) to detect the orthologs of H. arsenicoxydans genes and the comparative study of the genomes of three strains revealed that the 87.9 kb DNA fragment corresponding to HEAR1959 to HEAR2054 might be horizontally transferred to strain PD11. The 87.9 kb DNA fragment identified was found to contain three genes whose products were putatively involved in the metabolism of formaldehyde, a common intermediate of DCM and methanol. In addition, reverse transcription PCR analysis showed that all three genes were significantly expressed when strain PD11 was cultivated in the presence of DCM or methanol. These findings suggest that strain PD11 can effectively utilize the C1 compounds because of transfer of the mobile genetic elements from other bacterial species, for instance, from H. arsenicoxydans.


Subject(s)
Cupriavidus , Interspersed Repetitive Sequences , Methanol , Methylene Chloride , Methanol/metabolism , Cupriavidus/genetics , Cupriavidus/metabolism , Cupriavidus/drug effects , Methylene Chloride/metabolism , Interspersed Repetitive Sequences/genetics , Energy Metabolism/drug effects , Energy Metabolism/genetics , Genome, Bacterial/genetics , Gene Transfer, Horizontal
7.
Methods Mol Biol ; 2815: 79-91, 2024.
Article in English | MEDLINE | ID: mdl-38884912

ABSTRACT

Multidrug resistance, due to acquired antimicrobial resistance genes, is increasingly reported in the zoonotic pathogen Streptococcus suis. Most of these resistance genes are carried by chromosomal Mobile Genetic Elements (MGEs), in particular, Integrative and Conjugative Elements (ICEs) and Integrative and Mobilizable Elements (IMEs). ICEs and IMEs frequently form tandems or nested composite elements, which make their identification difficult. To evaluate their mobility, it is necessary to (i) select the suitable donor-recipient pairs for mating assays, (ii) do PCR excision tests to confirm that the genetic element is able to excise from the chromosome as a circular intermediate, and (iii) evaluate the transfer of the genetic element by conjugation by doing mating assays. In addition to a dissemination of resistance genes between S. suis strains, MGEs can lead to a spreading of resistance genes in the environment and toward pathogenic bacteria. This propagation had to be considered in a One Health perspective.


Subject(s)
Conjugation, Genetic , Interspersed Repetitive Sequences , Interspersed Repetitive Sequences/genetics , Gene Transfer, Horizontal , Streptococcus suis/genetics , Streptococcus suis/drug effects , Anti-Bacterial Agents/pharmacology , Drug Resistance, Bacterial/genetics , Polymerase Chain Reaction/methods , Genes, Bacterial
8.
Front Cell Infect Microbiol ; 14: 1368923, 2024.
Article in English | MEDLINE | ID: mdl-38694516

ABSTRACT

Introduction: Diagnosing Mycoplasma faucium poses challenges, and it's unclear if its rare isolation is due to infrequent occurrence or its fastidious nutritional requirements. Methods: This study analyzes the complete genome sequence of M. faucium, obtained directly from the pus of a sternum infection in a lung transplant patient using metagenomic sequencing. Results: Genome analysis revealed limited therapeutic options for the M. faucium infection, primarily susceptibility to tetracyclines. Three classes of mobile genetic elements were identified: two new insertion sequences, a new prophage (phiUMCG-1), and a species-specific variant of a mycoplasma integrative and conjugative element (MICE). Additionally, a Type I Restriction-Modification system was identified, featuring 5'-terminally truncated hsdS pseudogenes with overlapping repeats, indicating the potential for forming alternative hsdS variants through recombination. Conclusion: This study represents the first-ever acquisition of a complete circularized bacterial genome directly from a patient sample obtained from invasive infection of a primary sterile site using culture-independent, PCR-free clinical metagenomics.


Subject(s)
Genome, Bacterial , High-Throughput Nucleotide Sequencing , Metagenomics , Mycoplasma , Humans , Metagenomics/methods , Mycoplasma/genetics , Mycoplasma/isolation & purification , Mycoplasma/classification , Mycoplasma Infections/microbiology , Mycoplasma Infections/diagnosis , Whole Genome Sequencing/methods , Lung Transplantation , Prophages/genetics , Interspersed Repetitive Sequences/genetics , Anti-Bacterial Agents/pharmacology , Anti-Bacterial Agents/therapeutic use
9.
Appl Environ Microbiol ; 90(5): e0026424, 2024 May 21.
Article in English | MEDLINE | ID: mdl-38695519

ABSTRACT

The emergence of foodborne Salmonella strains carrying antimicrobial resistance (AMR) in mobile genetic elements (MGE) is a significant public health threat in a One Health context requiring continuous surveillance. Resistance to ciprofloxacin and cephalosporins is of particular concern. Since pigs are a relevant source of foodborne Salmonella for human beings, we studied transmissible AMR genes and MGE in a collection of 83 strains showing 9 different serovars and 15 patterns of multidrug resistant (MDR) previously isolated from pigs raised in the conventional breeding system of Northern Spain. All isolates were susceptible to ciprofloxacin and three isolates carried blaCMY-2 or blaCTX-M-9 genes responsible for cefotaxime resistance. Filter mating experiments showed that the two plasmids carrying blaCTX-M-9 were conjugative while that carrying blaCMY-2 was self-transmissible by transformation. Whole-genome sequencing and comparative analyses were performed on the isolates and plasmids. The IncC plasmid pSB109, carrying blaCMY-2, was similar to one found in S. Reading from cattle, indicating potential horizontal transfer between serovars and animal sources. The IncHI2 plasmids pSH102 in S. Heidelberg and pSTM45 in S. Typhimurium ST34, carrying blaCTX-M-9, shared similar backbones and two novel "complex class 1 integrons" containing different AMR and heavy metal genes. Our findings emphasize the importance of sequencing techniques to identify emerging AMR regions in conjugative and stable plasmids from livestock production. The presence of MGE carrying clinically relevant AMR genes raises public health concerns, requiring monitoring to mitigate the emergence of bacteria carrying AMR genes and subsequent spread through animals and food.IMPORTANCEThe emergence of foodborne Salmonella strains carrying antimicrobial resistance (AMR) in mobile genetic elements (MGE) is a significant public health threat in a One Health context. Since pigs are a relevant source of foodborne Salmonella for humans, in this study, we investigate different aspects of AMR in a collection of 83 Salmonella showing nine different serovars and 15 patterns of multidrug resistant (MDR) isolated from pigs raised in the conventional breeding system. Our findings emphasize the importance of sequencing techniques to identify emerging AMR regions in conjugative and stable plasmids from livestock production. The presence of MGE carrying clinically relevant AMR genes raises public health concerns, requiring monitoring to mitigate the emergence of bacteria carrying AMR genes and subsequent spread through animals and food.


Subject(s)
Anti-Bacterial Agents , Drug Resistance, Multiple, Bacterial , Interspersed Repetitive Sequences , Plasmids , Salmonella , Animals , Swine/microbiology , Plasmids/genetics , Salmonella/genetics , Salmonella/drug effects , Salmonella/isolation & purification , Drug Resistance, Multiple, Bacterial/genetics , Anti-Bacterial Agents/pharmacology , Humans , Cephalosporin Resistance/genetics , Salmonella Infections, Animal/microbiology , Spain , Swine Diseases/microbiology , Cephalosporins/pharmacology , Gene Transfer, Horizontal
10.
Nat Commun ; 15(1): 4555, 2024 May 29.
Article in English | MEDLINE | ID: mdl-38811529

ABSTRACT

Bacterial pathogens carrying multidrug resistance (MDR) plasmids are a major threat to human health. The acquisition of antibiotic resistance genes (ARGs) in plasmids is often facilitated by mobile genetic elements that copy or translocate ARGs between DNA molecules. The agglomeration of mobile elements in plasmids generates resistance islands comprising multiple ARGs. However, whether the emergence of resistance islands is restricted to specific MDR plasmid lineages remains understudied. Here we show that the agglomeration of ARGs in resistance islands is biased towards specific large plasmid lineages. Analyzing 6784 plasmids in 2441 Escherichia, Salmonella, and Klebsiella isolates, we quantify that 84% of the ARGs in MDR plasmids are found in resistance islands. We furthermore observe rapid evolution of ARG combinations in resistance islands. Most regions identified as resistance islands are shared among closely related plasmids but rarely among distantly related plasmids. Our results suggest the presence of barriers for the dissemination of ARGs between plasmid lineages, which are related to plasmid genetic properties, host range and the plasmid evolutionary history. The agglomeration of ARGs in plasmids is attributed to the workings of mobile genetic elements that operate within the framework of existing plasmid lineages.


Subject(s)
Anti-Bacterial Agents , Evolution, Molecular , Plasmids , Salmonella , Plasmids/genetics , Salmonella/genetics , Salmonella/drug effects , Anti-Bacterial Agents/pharmacology , Humans , Drug Resistance, Multiple, Bacterial/genetics , Klebsiella/genetics , Genomic Islands/genetics , Escherichia coli/genetics , Escherichia coli/drug effects , Interspersed Repetitive Sequences/genetics
11.
J Microbiol Methods ; 221: 106943, 2024 Jun.
Article in English | MEDLINE | ID: mdl-38705209

ABSTRACT

Bovine respiratory disease (BRD) is an important health and economic burden to the cattle industry worldwide. Three bacterial pathogens frequently associated with BRD (Mannheimia haemolytica, Pasteurella multocida, and Histophilus somni) can possess integrative and conjugative elements (ICEs), a diverse group of mobile genetic elements that acquire antimicrobial resistance (AMR) genes (ARGs) and decrease the therapeutic efficacy of antimicrobial drugs. We developed a duplex recombinase polymerase amplification (RPA) assay to detect up to two variants of ICEs in these Pasteurellaceae. Whole genome sequence analysis of M. haemolytica, P. multocida, and H. somni isolates harbouring ICEs revealed the presence of tnpA or ebrB next to tet(H), a conserved ARG that is frequently detected in ICEs within BRD-associated bacteria. This real-time multiplex RPA assay targeted both ICE variants simultaneously, denoted as tetH_tnpA and tetH_ebrB, with a limit of detection (LOD) of 29 (95% CI [23, 46]) and 38 genome copies (95% CI [30, 59]), respectively. DNA was extracted from 100 deep nasopharyngeal swabs collected from feedlot cattle on arrival. Samples were tested for ICEs using a real-time multiplex RPA assay, and for M. haemolytica, P. multocida, H. somni, and Mycoplasma bovis using both culture methods and RPA. The assay provided sensitive and accurate identification of ICEs in extracted DNA, providing a useful molecular tool for timely detection of potential risk factors associated with the development of antimicrobial-resistant BRD in feedlot cattle.


Subject(s)
Multiplex Polymerase Chain Reaction , Nasopharynx , Recombinases , Animals , Cattle , Nasopharynx/microbiology , Recombinases/genetics , Multiplex Polymerase Chain Reaction/methods , Multiplex Polymerase Chain Reaction/veterinary , Interspersed Repetitive Sequences/genetics , Cattle Diseases/microbiology , Cattle Diseases/diagnosis , DNA, Bacterial/genetics , Drug Resistance, Bacterial/genetics , Bovine Respiratory Disease Complex/microbiology , Conjugation, Genetic , Sensitivity and Specificity , Mannheimia haemolytica/genetics , Mannheimia haemolytica/isolation & purification , Pasteurellaceae/genetics , Pasteurellaceae/isolation & purification
12.
J Antimicrob Chemother ; 79(6): 1303-1308, 2024 Jun 03.
Article in English | MEDLINE | ID: mdl-38564255

ABSTRACT

BACKGROUND: Staphylococcus pseudintermedius is a common opportunistic pathogen of companion dogs and an occasional human pathogen. Treatment is hampered by antimicrobial resistance including methicillin resistance encoded by mecA within the mobile genetic element SCCmec. OBJECTIVES: SCCmec elements are diverse, especially in non-Staphyloccocus aureus staphylococci, and novel variants are likely to be present in S. pseudintermedius. The aim was to characterize the SCCmec elements found in four canine clinical isolates of S. pseudintermedius. MATERIAL AND METHODS: Isolates were whole-genome sequenced and SCCmec elements were assembled, annotated and compared to known SCCmec types. RESULTS AND DISCUSSION: Two novel SSCmec are present in these isolates. SCCmec7017-61515 is characterized by a novel combination of a Class A mec gene complex and a type 5 ccr previously only described in composite SCCmec elements. The other three isolates share a novel composite SCCmec with features of SCCmec types IV and VI. CONCLUSIONS: S. pseudintermedius is a reservoir of novel SSCmec elements that has implications for understanding antimicrobial resistant in veterinary and human medicine.


Subject(s)
Chromosomes, Bacterial , Dog Diseases , Methicillin Resistance , Staphylococcal Infections , Staphylococcus , Whole Genome Sequencing , Methicillin Resistance/genetics , Staphylococcus/genetics , Staphylococcus/drug effects , Staphylococcus/classification , Staphylococcus/isolation & purification , Animals , Dogs , Staphylococcal Infections/microbiology , Staphylococcal Infections/veterinary , Dog Diseases/microbiology , Chromosomes, Bacterial/genetics , Bacterial Proteins/genetics , Anti-Bacterial Agents/pharmacology , Microbial Sensitivity Tests , Genome, Bacterial , Genetic Variation , Interspersed Repetitive Sequences/genetics
13.
Nucleic Acids Res ; 52(10): 5496-5513, 2024 Jun 10.
Article in English | MEDLINE | ID: mdl-38686785

ABSTRACT

Cargo-mobilizing mobile elements (CMEs) are genetic entities that faithfully transpose diverse protein coding sequences. Although common in bacteria, we know little about eukaryotic CMEs because no appropriate tools exist for their annotation. For example, Starships are giant fungal CMEs whose functions are largely unknown because they require time-intensive manual curation. To address this knowledge gap, we developed starfish, a computational workflow for high-throughput eukaryotic CME annotation. We applied starfish to 2 899 genomes of 1 649 fungal species and found that starfish recovers known Starships with 95% combined precision and recall while expanding the number of annotated elements ten-fold. Extant Starship diversity is partitioned into 11 families that differ in their enrichment patterns across fungal classes. Starship cargo changes rapidly such that elements from the same family differ substantially in their functional repertoires, which are predicted to contribute to diverse biological processes such as metabolism. Many elements have convergently evolved to insert into 5S rDNA and AT-rich sequence while others integrate into random locations, revealing both specialist and generalist strategies for persistence. Our work establishes a framework for advancing mobile element biology and provides the means to investigate an emerging dimension of eukaryotic genetic diversity, that of genomes within genomes.


Subject(s)
Genome, Fungal , DNA Transposable Elements/genetics , Fungi/genetics , Genetic Variation , Genomics/methods , Interspersed Repetitive Sequences , Molecular Sequence Annotation , Phylogeny , Datasets as Topic
14.
J Hazard Mater ; 471: 134353, 2024 Jun 05.
Article in English | MEDLINE | ID: mdl-38678707

ABSTRACT

Aquatic microplastics (MPs) act as reservoirs for microbial communities, fostering the formation of a mobile resistome encompassing diverse antibiotic (ARGs) and biocide/metal resistance genes (BMRGs), and mobile genetic elements (MGEs). This collective genetic repertoire, referred to as the "plastiome," can potentially perpetuate environmental antimicrobial resistance (AMR). Our study examining two Japanese rivers near Tokyo revealed that waterborne MPs are primarily composed of polyethylene and polypropylene fibers and sheets of diverse origin. Clinically important genera like Exiguobacterium and Eubacterium were notably enriched on MPs. Metagenomic analysis uncovered a 3.46-fold higher enrichment of ARGs on MPs than those in water, with multidrug resistance genes (MDRGs) and BMRGs prevailing, particularly within MPs. Specific ARG and BMRG subtypes linked to resistance to vancomycin, beta-lactams, biocides, arsenic, and mercury showed selective enrichment on MPs. Network analysis revealed intense associations between host genera with ARGs, BMRGs, and MGEs on MPs, emphasizing their role in coselection. In contrast, river water exhibited weaker associations. This study underscores the complex interactions shaping the mobile plastiome in aquatic environments and emphasizes the global imperative for research to comprehend and effectively control AMR within the One Health framework.


Subject(s)
Microplastics , Rivers , Rivers/microbiology , Rivers/chemistry , Microplastics/toxicity , Anti-Bacterial Agents/pharmacology , Water Pollutants, Chemical/toxicity , Bacteria/genetics , Bacteria/drug effects , Water Microbiology , Interspersed Repetitive Sequences , Genes, Bacterial , Drug Resistance, Bacterial/genetics , Disinfectants/pharmacology , Microbiota/drug effects , Drug Resistance, Microbial/genetics
15.
J Hazard Mater ; 471: 134344, 2024 Jun 05.
Article in English | MEDLINE | ID: mdl-38678706

ABSTRACT

More information is needed to fully comprehend how acid mine drainage (AMD) affects the phototransformation of antibiotic resistant bacteria (ARB) and antibiotic resistance genes (ARGs) in karst water and sewage-irrigated farmland soil with abundant carbonate rocks (CaCO3) due to increasing pollution of AMD formed from pyrite (FeS2). The results showed FeS2 accelerated the inactivation of ARB with an inactivation of 8.7 log. Notably, extracellular and intracellular ARGs and mobile genetic elements (MGEs) also experienced rapid degradation. Additionally, the pH of the solution buffered by CaCO3 significantly influenced the photo-inactivation of ARB. The Fe2+ in neutral solution was present in Fe(II) coordination with strong reducing potential and played a crucial role in generating •OH (7.0 µM), which caused severe damage to ARB, ARGs, and MGEs. The •OH induced by photo-Fenton of FeS2 posed pressure to ARB, promoting oxidative stress response and increasing generation of reactive oxygen species (ROS), ultimately damaging cell membranes, proteins and DNA. Moreover, FeS2 contributed to a decrease in MIC of ARB from 24 mg/L to 4 mg/L. These findings highlight the importance of AMD in influencing karst water and sewage-irrigated farmland soil ecosystems. They are also critical in advancing the utilization of FeS2 to inactivate pathogenic bacteria.


Subject(s)
Calcium Carbonate , Iron , Mining , Sulfides , Calcium Carbonate/chemistry , Iron/chemistry , Sulfides/chemistry , Interspersed Repetitive Sequences , Drug Resistance, Microbial/genetics , Bacteria/genetics , Bacteria/drug effects , Genes, Bacterial , Drug Resistance, Bacterial/genetics , Anti-Bacterial Agents/pharmacology
16.
Sci Total Environ ; 926: 172115, 2024 May 20.
Article in English | MEDLINE | ID: mdl-38569972

ABSTRACT

Manure composting in traditional small-scale pig farms leads to the migration and diffusion of antibiotics and antibiotics resistance genes (ARGs) along the chain of transmission to the surrounding environment, increasing the risk of environmental resistance. Understanding the transmission patterns, driving factors, and health risks of ARGs on small-scale pig farms is important for effective control of ARGs transmission. This study was conducted on a small pig farm and its surrounding environment. The cross-media transmission of ARGs and their risks in the farming habitat were investigated using Metagenomic annotation and qPCR quantitative detection. The results indicate that ARGs in farms spread with manure pile-soil-channel sediment-mudflat sediment. Pig farm manure contributed 22.49 % of the mudflat sediment ARGs. Mobile genetic elements mediate the spread of ARGs across different media. Among them, tnpA and IS26 have the highest degree. Transmission of high-risk ARGs sul1 and tetM resulted in a 50 % and 116 % increase in host risk for sediment, respectively. This study provides a basis for farm manure management and control of the ARGs spread.


Subject(s)
Anti-Bacterial Agents , Genes, Bacterial , Animals , Swine , Farms , Anti-Bacterial Agents/pharmacology , Manure/analysis , Drug Resistance, Microbial/genetics , Interspersed Repetitive Sequences
17.
Environ Microbiol ; 26(4): e16630, 2024 Apr.
Article in English | MEDLINE | ID: mdl-38643972

ABSTRACT

Horizontal gene transfer (HGT) is a fundamental process in prokaryotic evolution, contributing significantly to diversification and adaptation. HGT is typically facilitated by mobile genetic elements (MGEs), such as conjugative plasmids and phages, which often impose fitness costs on their hosts. However, a considerable number of bacterial genes are involved in defence mechanisms that limit the propagation of MGEs, suggesting they may actively restrict HGT. In our study, we investigated whether defence systems limit HGT by examining the relationship between the HGT rate and the presence of 73 defence systems across 12 bacterial species. We discovered that only six defence systems, three of which were different CRISPR-Cas subtypes, were associated with a reduced gene gain rate at the species evolution scale. Hosts of these defence systems tend to have a smaller pangenome size and fewer phage-related genes compared to genomes without these systems. This suggests that these defence mechanisms inhibit HGT by limiting prophage integration. We hypothesize that the restriction of HGT by defence systems is species-specific and depends on various ecological and genetic factors, including the burden of MGEs and the fitness effect of HGT in bacterial populations.


Subject(s)
Bacteria , Gene Transfer, Horizontal , Gene Transfer, Horizontal/genetics , Bacteria/classification , Bacteria/genetics , Interspersed Repetitive Sequences/genetics , CRISPR-Cas Systems/genetics , Lysogeny/genetics , Species Specificity , Evolution, Molecular
18.
PLoS Pathog ; 20(4): e1012169, 2024 Apr.
Article in English | MEDLINE | ID: mdl-38640137

ABSTRACT

Integrative and conjugative elements (ICEs) play a vital role in bacterial evolution by carrying essential genes that confer adaptive functions to the host. Despite their importance, the mechanism underlying the stable inheritance of ICEs, which is necessary for the acquisition of new traits in bacteria, remains poorly understood. Here, we identified SezAT, a type II toxin-antitoxin (TA) system, and AbiE, a type IV TA system encoded within the ICESsuHN105, coordinately promote ICE stabilization and mediate multidrug resistance in Streptococcus suis. Deletion of SezAT or AbiE did not affect the strain's antibiotic susceptibility, but their duple deletion increased susceptibility, mainly mediated by the antitoxins SezA and AbiEi. Further studies have revealed that SezA and AbiEi affect the genetic stability of ICESsuHN105 by moderating the excision and extrachromosomal copy number, consequently affecting the antibiotic resistance conferred by ICE. The DNA-binding proteins AbiEi and SezA, which bind palindromic sequences in the promoter, coordinately modulate ICE excision and extracellular copy number by binding to sequences in the origin-of-transfer (oriT) and the attL sites, respectively. Furthermore, AbiEi negatively regulates the transcription of SezAT by binding directly to its promoter, optimizing the coordinate network of SezAT and AbiE in maintaining ICESsuHN105 stability. Importantly, SezAT and AbiE are widespread and conserved in ICEs harbouring diverse drug-resistance genes, and their coordinated effects in promoting ICE stability and mediating drug resistance may be broadly applicable to other ICEs. Altogether, our study uncovers the TA system's role in maintaining the genetic stability of ICE and offers potential targets for overcoming the dissemination and evolution of drug resistance.


Subject(s)
Bacterial Proteins , Streptococcus suis , Toxin-Antitoxin Systems , Streptococcus suis/genetics , Streptococcus suis/drug effects , Toxin-Antitoxin Systems/genetics , Bacterial Proteins/genetics , Bacterial Proteins/metabolism , Drug Resistance, Multiple, Bacterial/genetics , Streptococcal Infections/microbiology , Streptococcal Infections/genetics , Anti-Bacterial Agents/pharmacology , Conjugation, Genetic , Animals , Interspersed Repetitive Sequences
19.
PLoS One ; 19(4): e0301642, 2024.
Article in English | MEDLINE | ID: mdl-38683832

ABSTRACT

Horizontal gene transfer (HGT) is a well-documented strategy used by bacteria to enhance their adaptability to challenging environmental conditions. Through HGT, a group of conserved genetic elements known as mobile genetic elements (MGEs) is disseminated within bacterial communities. MGEs offer numerous advantages to the host, increasing its fitness by acquiring new functions that help bacteria contend with adverse conditions, including exposure to heavy metal and antibiotics. This study explores MGEs within microbial communities along the Yucatan coast using a metatranscriptomics approach. Prior to this research, nothing was known about the coastal Yucatan's microbial environmental mobilome and HGT processes between these bacterial communities. This study reveals a positive correlation between MGEs and antibiotic resistance genes (ARGs) along the Yucatan coast, with higher MGEs abundance in more contaminated sites. The Proteobacteria and Firmicutes groups exhibited the highest number of MGEs. It's important to highlight that the most abundant classes of MGEs might not be the ones most strongly linked to ARGs, as observed for the recombination/repair class. This work presents the first geographical distribution of the environmental mobilome in Yucatan Peninsula mangroves.


Subject(s)
Gene Transfer, Horizontal , Interspersed Repetitive Sequences , Microbiota , Interspersed Repetitive Sequences/genetics , Microbiota/genetics , Mexico , Bacteria/genetics , Bacteria/classification , Proteobacteria/genetics
20.
Nat Commun ; 15(1): 3477, 2024 Apr 24.
Article in English | MEDLINE | ID: mdl-38658529

ABSTRACT

Streptococcus dysgalactiae subspecies equisimilis (SDSE) and Streptococcus pyogenes share skin and throat niches with extensive genomic homology and horizontal gene transfer (HGT) possibly underlying shared disease phenotypes. It is unknown if cross-species transmission interaction occurs. Here, we conduct a genomic analysis of a longitudinal household survey in remote Australian First Nations communities for patterns of cross-species transmission interaction and HGT. Collected from 4547 person-consultations, we analyse 294 SDSE and 315 S. pyogenes genomes. We find SDSE and S. pyogenes transmission intersects extensively among households and show that patterns of co-occurrence and transmission links are consistent with independent transmission without inter-species interference. We identify at least one of three near-identical cross-species mobile genetic elements (MGEs) carrying antimicrobial resistance or streptodornase virulence genes in 55 (19%) SDSE and 23 (7%) S. pyogenes isolates. These findings demonstrate co-circulation of both pathogens and HGT in communities with a high burden of streptococcal disease, supporting a need to integrate SDSE and S. pyogenes surveillance and control efforts.


Subject(s)
Gene Transfer, Horizontal , Interspersed Repetitive Sequences , Streptococcal Infections , Streptococcus pyogenes , Streptococcus , Streptococcus pyogenes/genetics , Streptococcus pyogenes/isolation & purification , Streptococcus pyogenes/classification , Streptococcal Infections/transmission , Streptococcal Infections/microbiology , Humans , Streptococcus/genetics , Streptococcus/isolation & purification , Interspersed Repetitive Sequences/genetics , Australia , Genome, Bacterial/genetics , Female , Male , Child , Family Characteristics , Adult , Child, Preschool , Adolescent , Longitudinal Studies , Drug Resistance, Bacterial/genetics , Young Adult
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