posted on 2018-05-04, 10:56authored byNiran Patel, Alison Colyer, Steve Harris, Lucy Holcombe, Peter Andrew
Periodontal disease is one of the most important health concerns for companion animals. Research into canine forms of periodontitis has focused on the identification and characterization of the bacterial communities present. However, other microorganisms are known to inhabit the oral cavity and could also influence the disease process. A novel, broad spectrum 18S PCR was developed and used, in conjunction with next-generation sequencing analyses to target the identification of protists. Trichomonas sp. and Entamoeba sp. were identified from 92 samples of canine plaque. The overall prevalence of trichomonads was 56.52% (52/92) and entamoebae was 4.34% (4/92). Next-generation sequencing of pooled healthy, gingivitis, early-stage periodontitis, and severe periodontitis samples revealed the proportion of trichomonad sequences to be 3.51% (health), 2.84% (gingivitis), 6.07% (early periodontitis), and 35.04% (severe periodontitis), respectively, and entamoebae to be 0.01% (health), 0.01% (gingivitis), 0.80% (early-stage periodontitis), and 7.91% (severe periodontitis) respectively. Both genera of protists were statistically associated with plaque from dogs with periodontal disease. These findings provide the first conclusive evidence for the presence of oral protozoa in dog plaque and suggest a possible role for protozoa in the periodontal disease process.
Funding
We thank the WALTHAM Centre for Pet Nutrition for
providing funding and support for the project. We thank
the veterinarians and staff of Wey Referrals, England for
collecting plaque samples used in this study. We also
thank Dr Graham Clarke of the London School of Hygiene
and Tropical Medicine U.K. and Dr Simon Kilvington from
the University of Leicester U.K. for kindly providing protozoan
cultures and DNA.
History
Citation
Journal of Eukaryotic Microbiology, 2017, 64 (3), pp. 286-292
Author affiliation
/Organisation/COLLEGE OF LIFE SCIENCES/School of Medicine/Department of Infection, Immunity and Inflammation
Additional Supporting Information may be found online in
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Table S1. NCBI GenBank (Benson et al., 2013) protozoan
sequences chosen for sequence alignment during small
subunit rRNA PCR primer design.
Table S2. Small subunit rRNA gene amplicon sizes produced
from protozoa and other eukaryotic organisms using
the PCR developed in this study.
Table S3. Metadata associated with canine subgingival
plaque collections collected from animals presenting with
severe periodontal disease (stages 3–4), periodontal disease
stage 1, gingivitis, or from healthy animals.