Models for Gut-Mediated Horizontal Gene Transfer by Bacterial Plasmid Conjugation

Ott, Logan C.; Mellata, Melha

doi:10.3389/fmicb.2022.891548

Cited by 6 publications

(5 citation statements)

References 160 publications

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“…To mitigate the complexities of the in vivo gut environment and to overcome the limitations of in vitro models’ ability to account for complex host factors, we repeated conjugation assays in a chicken ceca explant model ( Ott and Mellata, 2022 ). Chicken ceca explants were collected from two-week-old chickens from a commercial farm and used as substrate in broth conjugation reactions using donor and recipient populations in the presence of ddH 2 O or 0.025 M of each SCFA ( Figure 4 ).…”

Section: Discussionmentioning

confidence: 99%

Short-chain fatty acids inhibit bacterial plasmid transfer through conjugation in vitro and in ex vivo chicken tissue explants

Ott,

Mellata

2024

Front. Microbiol.

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The animal gut acts as a potent reservoir for spreading and maintaining conjugative plasmids that confer antimicrobial resistance (AMR), fitness, and virulence attributes. Interventions that inhibit the continued emergence and expansion of AMR and virulent strains in agricultural and clinical environments are greatly desired. This study aims to determine the presence and efficacy of short-chain fatty acids (SCFA) inhibitory effects on the conjugal transfer of AMR plasmids. In vitro broth conjugations were conducted between donor Escherichia coli strains carrying AMP plasmids and the plasmid-less Escherichia coli HS-4 recipient strain. Conjugations were supplemented with ddH2O or SCFAs at 1, 0.1, 0.01, or 0.001 molar final concentration. The addition of SCFAs completely inhibited plasmid transfer at 1 and 0.1 molar and significantly (p < 0.05) reduced transfer at 0.01 molar, regardless of SCFA tested. In explant models for the chicken ceca, either ddH2O or a final concentration of 0.025 M SCFAs were supplemented to the explants infected with donor and recipient E. coli. In every SCFA tested, significant decreases in transconjugant populations compared to ddH2O-treated control samples were observed with minimal effects on donor and recipient populations. Finally, significant reductions in transconjugants for plasmids of each incompatibility type (IncP1ε, IncFIβ, and IncI1) tested were detected. This study demonstrates for the first time the broad inhibition ability of SCFAs on bacterial plasmid transfer and eliminates AMR with minimal effect on bacteria. Implementing interventions that increase the concentrations of SCFAs in the gut may be a viable method to reduce the risk, incidence, and rate of AMR emergence in agricultural and human environments.

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Section: Discussionmentioning

confidence: 99%

Short-chain fatty acids inhibit bacterial plasmid transfer through conjugation in vitro and in ex vivo chicken tissue explants

Ott,

Mellata

2024

Front. Microbiol.

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“…T4SS facilitate the transport of DNA and proteins either directly into the environment or into target cells [ 76 ]. T4SS-mediated horizontal DNA transfer, also known as conjugation, spreads mobile genetic elements between bacteria, and is a key process for bacterial evolution and for antibiotic-resistant gene transfer [ 77 ]. Another important function of T4SS is the injection of antibacterial proteins directly into neighboring cells.…”

Section: Discussionmentioning

confidence: 99%

Genetic Basis and Expression Pattern Indicate the Biocontrol Potential and Soil Adaption of Lysobacter capsici CK09

Yang

Yuan

et al. 2023

Microorganisms

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Lysobacter species have attracted increasing attention in recent years due to their capacities to produce diverse secondary metabolites against phytopathogens. In this research, we analyzed the genomic and transcriptomic patterns of Lysobacter capsici CK09. Our data showed that L. capsici CK09 harbored various contact-independent biocontrol traits, such as fungal cell wall lytic enzymes and HSAF/WAP-8294A2 biosynthesis, as well as several contact-dependent machineries, including type 2/4/6 secretion systems. Additionally, a variety of hydrolytic enzymes, particularly extracellular enzymes, were found in the L. capsici CK09 genome and predicted to improve its adaption in soil. Furthermore, several systems, including type 4 pili, type 3 secretion system and polysaccharide biosynthesis, can provide a selective advantage to L. capsici CK09, enabling the species to live on the surface in soil. The expression of these genes was then confirmed via transcriptomic analysis, indicating the activities of these genes. Collectively, our research provides a comprehensive understanding of the biocontrol potential and soil adaption of L. capsici CK09 and implies the potential of this strain for application in the future.

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“…using suitable recipients expressing the correct receptors, will increase conjugation frequencies. Furthermore, the increase in abundance of data from in situ, in vivo , and in vitro studies will benefit the development of in silico models used to simulate rate and incidence of conjugation in the human gut, as reviewed by Ott and Mellata ( 2022 ). The continued development of the discussed approaches will in turn further our collective understanding of the significance of the human gut microbiome on the dynamics of AMR plasmids, and their contribution to the emergence and prevalence of clinically important pathogens.…”

Section: Discussionmentioning

confidence: 99%

In situ, in vivo, and in vitro approaches for studying AMR plasmid conjugation in the gut microbiome

Kessler

Hou

Neo

et al. 2022

FEMS Microbiology Reviews

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Antimicrobial resistance (AMR) is a global threat, with evolution and spread of resistance to frontline antibiotics outpacing the development of novel treatments. The spread of AMR is perpetuated by transfer of antimicrobial resistance genes (ARGs) between bacteria, notably those encoded by conjugative plasmids. The human gut microbiome is a known ‘melting pot’ for plasmid conjugation, with ARG transfer in this environment widely documented. There is a need to better understand the factors affecting the incidence of these transfer events, and to investigate methods of potentially counteracting the spread of ARGs. This review describes the use and potential of three approaches to studying conjugation in the human gut: observation of in situ events in hospitalized patients, modelling of the microbiome in vivo predominantly in rodent models, and the use of in vitro models of various complexities. Each has brought unique insights to our understanding of conjugation in the gut. The use and development of these systems, and combinations thereof, will be pivotal in better understanding the significance, prevalence and manipulability of horizontal gene transfer in the gut microbiome.

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Models for Gut-Mediated Horizontal Gene Transfer by Bacterial Plasmid Conjugation

Cited by 6 publications

References 160 publications

Short-chain fatty acids inhibit bacterial plasmid transfer through conjugation in vitro and in ex vivo chicken tissue explants

Short-chain fatty acids inhibit bacterial plasmid transfer through conjugation in vitro and in ex vivo chicken tissue explants

Genetic Basis and Expression Pattern Indicate the Biocontrol Potential and Soil Adaption of Lysobacter capsici CK09

In situ, in vivo, and in vitro approaches for studying AMR plasmid conjugation in the gut microbiome

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