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Cyromazine accelerates the dissemination of antibiotic resistance genes by promoting the conjugative transfer of the SXT integrative conjugative element

ABSTRACT:

Antimicrobial resistance (AMR) is one of the most urgent challenges in public health, and the horizontal transfer of antibiotic resistance genes (ARGs) mediated by mobile genetic elements (MGEs) drives its widespread global dissemination. Although integrative conjugative elements (ICEs) outnumber conjugative plasmids, research on ICE-mediated horizontal gene transfer (HGT) remains largely lacking, especially concerning SXT ICEs, which are highly prevalent, pose significant risks, and are directly related to human health. Cyromazine is a widely used feed additive, with high detection rates in the feces of farmed animals and related environments. It has also been shown to increase the abundance of ARGs, and the potential driving mechanisms urgently require further exploration. Thus, we investigated the impact of cyromazine on SXT ICE-mediated ARGs transfer and elucidated its underlying mechanisms. Our study demonstrates that cyromazine promotes SXT ICE conjugative transfer both intra- and inter-species. This facilitating effect confirmed in mouse intraperitoneal and intestinal conjugation models. The potential mechanisms include cyromazine-induced ROS accumulation and triggering of the SOS response, which could promote SXT ICE excision and activate conjugation-related operons. Furthermore, enhanced energy metabolism and altered lipid metabolism further facilitate SXT ICE transfer. Collectively, our study partially fills the gap in understanding ICE-mediated AMR transmission dynamics, and highlighting that the exacerbated AMR crisis is associated with cyromazine-stimulated high-frequency SXT ICE conjugation in vitro and in vivo.

INSTRUMENT(S): Liquid Chromatography MS - negative - reverse phase, Liquid Chromatography MS - positive - reverse phase

PROVIDER: MTBLS13146 | MetaboLights | 2025-10-15

REPOSITORIES: MetaboLights

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			Action	DRS
	a_MTBLS13146_LC-MS_negative_reverse-phase_metabolite_profiling.txt	Txt
	a_MTBLS13146_LC-MS_positive_reverse-phase_metabolite_profiling.txt	Txt
	i_Investigation.txt	Txt
	m_MTBLS13146_LC-MS_negative_reverse-phase_metabolite_profiling_v2_maf.tsv	Tabular
	m_MTBLS13146_LC-MS_positive_reverse-phase_metabolite_profiling_v2_maf.tsv	Tabular

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Project description:<p>The study of antimicrobial resistance (AMR) in infectious diarrhea has generally been limited to cultivation, antimicrobial susceptibility testing and targeted PCR assays. When individual strains of significance are identified, whole genome shotgun (WGS) sequencing of important clones and clades is performed. Genes that encode resistance to antibiotics have been detected in environmental, insect, human and animal metagenomes and are known as "resistomes". While metagenomic datasets have been mined to characterize the healthy human gut resistome in the Human Microbiome Project and MetaHIT and in a Yanomani Amerindian cohort, directed metagenomic sequencing has not been used to examine the epidemiology of AMR. Especially in developing countries where sanitation is poor, diarrhea and enteric pathogens likely serve to disseminate antibiotic resistance elements of clinical significance. Unregulated use of antibiotics further exacerbates the problem by selection for acquisition of resistance. This is exemplified by recent reports of multiple antibiotic resistance in Shigella strains in India, in Escherichia coli in India and Pakistan, and in nontyphoidal Salmonella (NTS) in South-East Asia. We propose to use deep metagenomic sequencing and genome level assembly to study the epidemiology of AMR in stools of children suffering from diarrhea. Here the epidemiology component will be surveillance and analysis of the microbial composition (to the bacterial species/strain level where possible) and its constituent antimicrobial resistance genetic elements (such as plasmids, integrons, transposons and other mobile genetic elements, or MGEs) in samples from a cohort where diarrhea is prevalent and antibiotic exposure is endemic. The goal will be to assess whether consortia of specific mobile antimicrobial resistance elements associate with species/strains and whether their presence is enhanced or amplified in diarrheal microbiomes and in the presence of antibiotic exposure. This work could potentially identify clonal complexes of organisms and MGEs with enhanced resistance and the potential to transfer this resistance to other enteric pathogens.</p> <p>We have performed WGS, metagenomic assembly and gene/protein mapping to examine and characterize the types of AMR genes and transfer elements (transposons, integrons, bacteriophage, plasmids) and their distribution in bacterial species and strains assembled from DNA isolated from diarrheal and non-diarrheal stools. The samples were acquired from a cohort of pediatric patients and controls from Colombia, South America where antibiotic use is prevalent. As a control, the distribution and abundance of AMR genes can be compared to published studies where resistome gene lists from healthy cohort sequences were compiled. Our approach is more epidemiologic in nature, as we plan to identify and catalogue antimicrobial elements on MGEs capable of spread through a local population and further we will, where possible, link mobile antimicrobial resistance elements with specific strains within the population.</p>

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Cyromazine accelerates the dissemination of antibiotic resistance genes by promoting the conjugative transfer of the SXT integrative conjugative element

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