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External Ear Canal Evaluation in Dogs with Chronic Suppurative Otitis Externa: Comparison of Direct Cytology, Bacterial Culture and 16S Amplicon Profiling

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A discrepancy between cytology and bacterial culture methods is sometimes observed in canine otitis externa. The objective of this study was to compare results from cytology, bacterial culture and 16S amplicon profiling. Twenty samples from 16 dogs with chronic suppurative otitis externa were collected. A direct cytological evaluation was carried out during the consultations. Aerobic bacterial culture and susceptibility were performed by an external laboratory used in routine practice. For 16S amplicon profiling, DNA was extracted and the hypervariable segment V1–V3 of the 16S rDNA was amplified and then sequenced with a MiSeq Illumina sequence carried out by the Mothur software using the SILVA database. A good correlation between cytology and bacterial culture was observed in 60% of the samples. Some bacterial species revealed by bacterial culture were present with low relative abundance (<10%) in 16S amplicon profiling. Some bacterial species revealed by the 16S amplicon profiling analysis were not identified with culture; most of the time, the offending species was a Corynebacterium. To conclude, a careful interpretation of the results of bacterial culture should be made and always be in agreement with the cytology. The 16S amplicon profiling method appears to be a more sensitive method for detecting strains present in suppurative otitis but does not provide information on bacterial susceptibility.
Title: External Ear Canal Evaluation in Dogs with Chronic Suppurative Otitis Externa: Comparison of Direct Cytology, Bacterial Culture and 16S Amplicon Profiling
Description:
A discrepancy between cytology and bacterial culture methods is sometimes observed in canine otitis externa.
The objective of this study was to compare results from cytology, bacterial culture and 16S amplicon profiling.
Twenty samples from 16 dogs with chronic suppurative otitis externa were collected.
A direct cytological evaluation was carried out during the consultations.
Aerobic bacterial culture and susceptibility were performed by an external laboratory used in routine practice.
For 16S amplicon profiling, DNA was extracted and the hypervariable segment V1–V3 of the 16S rDNA was amplified and then sequenced with a MiSeq Illumina sequence carried out by the Mothur software using the SILVA database.
A good correlation between cytology and bacterial culture was observed in 60% of the samples.
Some bacterial species revealed by bacterial culture were present with low relative abundance (<10%) in 16S amplicon profiling.
Some bacterial species revealed by the 16S amplicon profiling analysis were not identified with culture; most of the time, the offending species was a Corynebacterium.
To conclude, a careful interpretation of the results of bacterial culture should be made and always be in agreement with the cytology.
The 16S amplicon profiling method appears to be a more sensitive method for detecting strains present in suppurative otitis but does not provide information on bacterial susceptibility.

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