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Published on: January 17, 2014
Characterization of nontypeable Actinobacillus pleuropneumoniae isolates
Ho To1,2, Kaho Teshima1,2, Michiha Kon1,2
1Nippon Institute for Biological Science, Ome, Tokyo, Japan (To, Teshima, Kon, Yasuda, Akaike, Shibuya, Nagai, Sasakawa).
Abstract:
Two Actinobacillus pleuropneumoniae isolates from clinical cases of porcine pleuropneumonia in Japan were positive in the capsular serovar 15-specific PCR assay, but nontypeable (NT) in the agar gel precipitation (AGP) test. Nucleotide sequence analysis of gene clusters involved in the biosynthesis of capsular polysaccharide (CPS) and lipopolysaccharide O-polysaccharide (O-PS) revealed that both clusters contained transposable element ISApl1 of A. pleuropneumoniae belonging to the IS30 family. Immunoblot analysis revealed that these 2 isolates could not produce O-PS. We conclude that the ISApl1 of A. pleuropneumoniae can interfere in the biosynthesis of both CPS and O-PS.
Insights
Transposable elements like IS Apl1 in Actinobacillus pleuropneumoniae can disrupt capsule and O-polysaccharide production. This finding explains why some bacterial isolates appear nontypeable despite testing positive for specific genes.
Area of Science:
- Veterinary Microbiology
- Bacterial Genetics
- Swine Disease Research
Background:
- Porcine pleuropneumonia, caused by Actinobacillus pleuropneumoniae, is a significant swine respiratory disease.
- Accurate serotyping is crucial for diagnosing and controlling outbreaks.
- Some isolates present challenges in traditional typing methods.
Purpose of the Study:
- To investigate the genetic basis for atypical serotyping results in Actinobacillus pleuropneumoniae isolates.
- To determine the role of transposable elements in capsule and O-polysaccharide biosynthesis.
- To understand the implications for bacterial typing and pathogenesis.
Main Methods:
- Polymerase Chain Reaction (PCR) for capsular serovar 15-specific gene detection.
- Agar Gel Precipitation (AGP) test for serotyping.
- Nucleotide sequence analysis of polysaccharide biosynthesis gene clusters.
- Immunoblot analysis to assess O-polysaccharide production.
Main Results:
- Two clinical isolates were PCR-positive for serovar 15 but AGP-negative (nontypeable).
- Sequence analysis revealed the presence of the transposable element IS Apl1 in polysaccharide biosynthesis gene clusters of both isolates.
- Immunoblotting confirmed the absence of O-polysaccharide production in these isolates.
- The IS Apl1 element was implicated in the disruption of both capsular polysaccharide (CPS) and lipopolysaccharide O-polysaccharide (O-PS) biosynthesis.
Conclusions:
- The transposable element IS Apl1 of Actinobacillus pleuropneumoniae can interfere with the biosynthesis of both CPS and O-PS.
- This interference can lead to nontypeable isolates despite the presence of specific serotype genes.
- Understanding these genetic mechanisms is vital for accurate diagnostics and effective disease management in swine.
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