Macrolide inactivation gene cluster mphA-mrx-mphR adjacent to a class 1 integron in Aeromonas hydrophila isolated from a diarrhoeic pig in Oklahoma

Poole, Toni L.; Callaway, Todd R.; Bischoff, Kenneth M.; Warnes, Carl E.; Nisbet, David J.

doi:10.1093/jac/dki421

Cited by 38 publications

(36 citation statements)

References 41 publications

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“…BLAST identified similarity to a region on the 48-kb plasmid pRSB101 (NC_006385) from an uncultured host, including a homolog of chrA resembling a chromate ion transporter, an ORF of unknown function, an operon carrying a macrolide phosphotransferase (mphA) and its repressor (mphR), and two insertion sequence (IS) elements. The macrolide resistance operon was previously observed inserted into the integron of a Tn21-like transposon in a strain of Aeromonas hydrophila isolated from swine (25). This integron also contained the dfrA12, orfF, and aadA2 cassettes identified on the pLEW517 transposon; however, it lacked the chrA homolog, the IS elements, and the ORF of unknown function.…”

Section: Resultsmentioning

confidence: 99%

Facile Recovery of Individual High-Molecular-Weight, Low-Copy-Number Natural Plasmids for Genomic Sequencing

Williams

Detter

Barry

et al. 2006

Appl Environ Microbiol

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Sequencing of the large (>50 kb), low-copy-number (<5 per cell) plasmids that mediate horizontal gene transfer has been hindered by the difficulty and expense of isolating DNA from individual plasmids of this class. We report here that a kit method previously devised for purification of bacterial artificial chromosomes (BACs) can be adapted for effective preparation of individual plasmids up to 220 kb from wild gram-negative and gram-positive bacteria. Individual plasmid DNA recovered from less than 10 ml of Escherichia coli, Staphylococcus, and Corynebacterium cultures was of sufficient quantity and quality for construction of highcoverage libraries, as shown by sequencing five native plasmids ranging in size from 30 kb to 94 kb. We also report recommendations for vector screening to optimize plasmid sequence assembly, preliminary annotation of novel plasmid genomes, and insights on mobile genetic element biology derived from these sequences. Adaptation of this BAC method for large plasmid isolation removes one major technical hurdle to expanding our knowledge of the natural plasmid gene pool.Recent genome sequencing and analysis has revealed extensive horizontal gene transfer among bacterial genomes (4,14). Remnants of mobile genetic elements (MGEs), such as plasmids and bacteriophages (23), are often found adjacent to horizontally transferred chromosomal regions, indicating that these elements are important mediators of gene transfer between bacterial chromosomes. The MGEs themselves also typically carry genes for virulence factors, antibiotic resistances, and novel metabolic processes that enable bacterial hosts to adapt to new environmental conditions (11,12).Despite the recognized importance of these elements, genomic analysis of MGEs has been limited. Whereas the total size of sequenced bacterial genomes is 1.3 Gb, only 61 Mb of plasmid genomes and 30 Mb of phage genomes have been sequenced previously (11). Most MGE sequences have been obtained fortuitously during sequencing of their hosts' genomes, resulting in a bias towards MGEs associated with a limited selection of organisms. Large (Ͼ50 kb), conjugative plasmids are especially poorly represented in current sequence databases, constituting only 20% of all plasmid sequences in GenBank at present. Most commercial kits for plasmid DNA preparation are designed for small, high-copy-number plasmids. The traditional methods of high-molecular-weight plasmid isolation, such as cesium chloride density gradient centrifugation (26) and pulsed-field gel electrophoresis, require equipment and expertise that are not widely available. Moreover, these and other techniques, such as Eckhardt in-well lysis (6), are time and labor intensive and thus unsuitable for a high-throughput approach.In the course of large-scale sequencing projects such as the Human Genome Project, magnetic beads were modified for purification of nucleic acids, including bacterial artificial chromosome (BAC) clones (7,15,28). The method used here, termed Solid-Phase Reversible Immobilization (SPRI)...

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Section: Resultsmentioning

confidence: 99%

Facile Recovery of Individual High-Molecular-Weight, Low-Copy-Number Natural Plasmids for Genomic Sequencing

Williams

Detter

Barry

et al. 2006

Appl Environ Microbiol

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“…Similar findings were also reported in Pseudomonas aeruginosa (Jeong et al 2009;Colinon et al 2010). The link between the qac genes and macrolide inactivation genes was revealed in Aeromonas hydrophila (Poole et al 2006) as well as in microflora from a wastewater treatment plant (Szczepanowski et al 2004). Therefore, the use of various cationic biocides may be also responsible for the selection of bacteria resistant to antibiotics (Russell 2000;Hegstad et al 2010).…”

Section: The Qac Genes and Resistance To Antibioticsmentioning

confidence: 99%

Genetic basis of resistance to quaternary ammonium compounds - the qac genes and their role: a review

Jaglič¹,

Cervinkova²

2012

Vet. Med.

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ABSTRACT:Although the qac genes are named after one of their main substrates (i.e., quaternary ammonium compounds), these genes also code for resistance to a broad spectrum of other cationic compounds such as intercalating dyes, diamidines and biguanides. The various Qac proteins are involved in relatively low specific effluxbased multidrug pumps and belong to a family of small multidrug resistance proteins. Even though the practical significance of qac-mediated resistance lies mainly in resistance to antiseptics, contradictory findings on this issue are still reported. Therefore, the aim of this review is to summarise the current knowledge on qac-mediated resistance with special emphasis on resistance to antiseptics and its relevance for practice.

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“…The combination of dfrA14 and recombined aadA6 cassettes has previously been detected in E. coli of porcine origin (6). The combination dfrA12-orfF-aadA2 has been seen in Salmonella Typhimurium from imported seafood (7), Salmonella enterica serovar Schwarzengrund from chicken (22), and A. hydrophila (15) or E. coli (6) from swine. The combination dfrB1-aadA1-catB2 was seen as part of larger bla VIM-1 -carrying integron structures in Klebsiella pneumoniae (19) and E. coli (20).…”

mentioning

confidence: 99%

Molecular Basis of Sulfonamide and Trimethoprim Resistance in Fish-Pathogenic Aeromonas Isolates

Kadlec

Czapiewski

Kaspar

et al. 2011

Appl Environ Microbiol

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Macrolide inactivation gene cluster mphA-mrx-mphR adjacent to a class 1 integron in Aeromonas hydrophila isolated from a diarrhoeic pig in Oklahoma

Cited by 38 publications

References 41 publications

Facile Recovery of Individual High-Molecular-Weight, Low-Copy-Number Natural Plasmids for Genomic Sequencing

Facile Recovery of Individual High-Molecular-Weight, Low-Copy-Number Natural Plasmids for Genomic Sequencing

Genetic basis of resistance to quaternary ammonium compounds - the qac genes and their role: a review

Molecular Basis of Sulfonamide and Trimethoprim Resistance in Fish-Pathogenic Aeromonas Isolates

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