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ICEApl1, an Integrative Conjugative Element Related to ICEHin1056, Identified in the Pig Pathogen Actinobacillus

Janine T Bossé¹, Yanwen Li¹, Roberto Fernandez Crespo¹

¹Section of Paediatrics, Department of Medicine, Imperial College London London, UK.

Frontiers in Microbiology

|July 6, 2016

Abstract:

ICEApl1 was identified in the whole genome sequence of MIDG2331, a tetracycline-resistant (MIC = 8 mg/L) serovar 8 clinical isolate of Actinobacillus pleuropneumoniae, the causative agent of porcine pleuropneumonia. PCR amplification of virB4, one of the core genes involved in conjugation, was used to identify other A. pleuropneumoniae isolates potentially carrying ICEApl1. MICs for tetracycline were determined for virB4 positive isolates, and shotgun whole genome sequence analysis was used to confirm presence of the complete ICEApl1. The sequence of ICEApl1 is 56083 bp long and contains 67 genes including a Tn10 element encoding tetracycline resistance. Comparative sequence analysis was performed with similar integrative conjugative elements (ICEs) found in other members of the Pasteurellaceae. ICEApl1 is most similar to the 59393 bp ICEHin1056, from Haemophilus influenzae strain 1056. Although initially identified only in serovar 8 isolates of A. pleuropneumoniae (31 from the UK and 1 from Cyprus), conjugal transfer of ICEApl1 to representative isolates of other serovars was confirmed. All isolates carrying ICEApl1 had a MIC for tetracycline of 8 mg/L. This is, to our knowledge, the first description of an ICE in A. pleuropneumoniae, and the first report of a member of the ICEHin1056 subfamily in a non-human pathogen. ICEApl1 confers resistance to tetracycline, currently one of the more commonly used antibiotics for treatment and control of porcine pleuropneumonia.

Insights

A novel integrative conjugative element, ICEApl1, was discovered in Actinobacillus pleuropneumoniae, conferring tetracycline resistance. This finding is crucial for understanding and controlling porcine pleuropneumonia.

Area of Science:

Microbiology
Genetics
Veterinary Medicine

Background:

Actinobacillus pleuropneumoniae causes porcine pleuropneumonia, a significant disease in swine.
Tetracycline is commonly used for treating and controlling this disease.
Integrative conjugative elements (ICEs) are mobile genetic elements that can transfer between bacteria.

Purpose of the Study:

To identify and characterize novel mobile genetic elements in Actinobacillus pleuropneumoniae.
To investigate the genetic basis of tetracycline resistance in A. pleuropneumoniae.
To determine the potential for horizontal gene transfer of antibiotic resistance.

Main Methods:

Whole genome sequencing of clinical isolates.
Polymerase Chain Reaction (PCR) amplification of specific genes (virB4).
Determination of Minimum Inhibitory Concentrations (MICs) for tetracycline.
Comparative sequence analysis of ICEs.

Main Results:

Discovery of ICEApl1, a 56083 bp integrative conjugative element in A. pleuropneumoniae.
ICEApl1 contains 67 genes, including a Tn10 element conferring tetracycline resistance.
ICEApl1 was found in serovar 8 isolates and successfully transferred to other serovars.
All ICEApl1-carrying isolates exhibited a tetracycline MIC of 8 mg/L.

Conclusions:

This is the first description of an ICE in A. pleuropneumoniae.
ICEApl1 represents a new member of the ICEHin1056 subfamily, previously found only in human pathogens.
ICEApl1 confers clinically relevant tetracycline resistance, impacting treatment strategies for porcine pleuropneumonia.

Read full analysis

Keywords:

Pasteurellaceae animal infections antibiotic resistance conjugation respiratory tract

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