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Author Spotlight: Advancing Antibiotic Resistance Research Using an Efflux-Deficient Bacterial Strain and a Single-Copy Gene Expression System
Published on: January 5, 2024
A large conjugative Acinetobacter baumannii plasmid carrying the sul2 sulphonamide and strAB streptomycin resistance
Mohammad Hamidian1, Stephanie J Ambrose1, Ruth M Hall1
1School of Life and Environmental Sciences, The University of Sydney, NSW 2006, Australia.
Abstract:
Acinetobacter baumannii is an important nosocomial pathogen that often complicates treatment because of its high level of resistance to antibiotics. Though plasmids can potentially introduce various genes into bacterial strains, compared to other Gram-negative bacteria, information about the unique A. baumannii plasmid repertoire is limited. Here, whole genome sequence data was used to determine the plasmid content of strain A297 (RUH875), the reference strain for the globally disseminated multiply resistant A. baumannii clone, global clone 1(GC1). A297 contains three plasmids. Two known plasmids were present; one, pA297-1 (pRAY*), carries the aadB gentamicin, kanamycin and tobramycin resistance gene and another is an 8.7kb cryptic plasmid often found in GC1 isolates. The third plasmid, pA297-3, is 200kb and carries the sul2 sulphonamide resistance gene and strAB streptomycin resistance gene within Tn6172 and a mer mercuric ion resistance module elsewhere. pA297-3 transferred sulphonamide, streptomycin and mercuric ion resistance at high frequency to a susceptible A. baumannii recipient, and contains several genes potentially involved in conjugative transfer. However, a relaxase gene was not found. It also includes several genes encoding proteins involved in DNA metabolism such as partitioning. However, a gene encoding a replication initiation protein could not be found. pA297-3 includes two copies of a Miniature Inverted-Repeat Transposable Element (MITE), named MITE-297, bracketing a 77.5kb fragment, which contains several IS and the mer module. Several plasmids related to but smaller than pA297-3 were found in the GenBank nucleotide database. They were found in different A. baumannii clones and are wide spread. They all contain either Tn6172 or a variant in the same position in the backbone as Tn6172 in pA297-3. Some related plasmids have lost the segment between the MITE-297 copies and retain only one MITE-297. Others have segments of various lengths between two MITE-297 copies, and these can be derived from the region in pA297-3 via a deletion adjacent to IS related to IS26 such as IS1007 or IS1007-like. pA297-3 and its relatives represent a third type of conjugative Acinetobacter plasmid that contributes to the dissemination of antibiotic resistance in this species.
Insights
This study characterizes novel plasmids in Acinetobacter baumannii, revealing a new type of conjugative plasmid, pA297-3. This plasmid facilitates the spread of antibiotic resistance genes, including sul2 and strAB, and mercuric ion resistance.
Area of Science:
- Microbiology
- Genetics
- Molecular Biology
Background:
- Acinetobacter baumannii is a significant nosocomial pathogen known for its extensive antibiotic resistance.
- Understanding the plasmid content of A. baumannii is crucial for combating resistance, yet information remains limited compared to other Gram-negative bacteria.
Purpose of the Study:
- To determine the plasmid content of the globally disseminated, multiply resistant A. baumannii strain A297 (RUH875), the reference strain for Global Clone 1 (GC1).
- To characterize a novel, large plasmid (pA297-3) and its role in the dissemination of antibiotic resistance genes.
Main Methods:
- Whole genome sequencing of A. baumannii strain A297.
- Plasmid identification and characterization, including size, gene content, and presence of mobile genetic elements.
- Conjugation experiments to assess the transferability of resistance genes.
Main Results:
- Strain A297 harbors three plasmids: pA297-1 (carrying aadB), an 8.7kb cryptic plasmid common in GC1, and a novel 200kb plasmid, pA297-3.
- pA297-3 contains the sul2 and strAB resistance genes within Tn6172 and a mercuric ion resistance module.
- pA297-3 efficiently transferred sulfonamide, streptomycin, and mercuric ion resistance to a susceptible A. baumannii recipient and contains genes suggestive of conjugative transfer.
- pA297-3 features two copies of a Miniature Inverted-Repeat Transposable Element (MITE), MITE-297, flanking a large fragment containing mobile genetic elements and the mercury resistance module.
- Related plasmids, smaller than pA297-3, were identified in other A. baumannii clones, indicating widespread dissemination of this plasmid type.
Conclusions:
- pA297-3 represents a third type of conjugative Acinetobacter plasmid, distinct from previously described types.
- This novel plasmid type plays a significant role in the dissemination of antibiotic and heavy metal resistance genes within A. baumannii populations.
- The structural organization of pA297-3, including MITEs and mobile genetic elements, likely contributes to its stability and transferability, driving antimicrobial resistance spread.
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