| Periodontal disease(PD)in dogs is a very common clinical infectious disease.The main clinical manifestations are gingivitis and periodontitis,which are often related to the breed,age,and living habits of dogs.Can cause disease in other organs or systems.PD is mainly caused by bacteria colonizing subgingival plaque,but the bacterial colonization of subgingival plaque and its dynamic changes are unclear.In this paper,by analyzing the canine cases of periodontal disease,and analyzing the composition of subgingival flora and its bacterial diversity.The purpose of this paper is to understand the composition of canine subgingival bacteria more clearly,to further deepen the understanding of the pathogenesis and characteristics of canine periodontal disease.understand,and to provide a scientific basis for clinical prevention and treatment of the disease.The results of this study are as follows:1.A total of 91 canine cases of periodontal disease were obtained in this paper,including 42 cases of gingivitis and 49 cases of periodontitis.The periodontal indexspecific condition scores(including gingival index and alveolar bleeding index)of canine periodontitis were significantly higher than those of gingivitis(p<0.001),and the plaque and calculus index scores were also significantly higher than those of gingivitis(p<0.001).2.Association of PD with dog breed,sex and age.By analyzing the basic case information of 91 affected dogs,it was found that the proportion of mongrel dogs was the highest(44/91,48.35%),followed by poodles(16/91,17.58%).Analysis of the sex ratio of affected dogs found that the number of male dogs was slightly higher than that of female dogs,1.17:1.Affected dogs can develop gingivitis at about 2 years old,while periodontitis mainly occurs in dogs over 4 years old,of which 75.51% of dogs are over 6years old.3.The collected subgingival samples of 111 dogs(20 healthy samples,42 gingivitis samples,49 periodontitis samples)were cultured aerobic and anaerobic,respectively,and identified by colony morphology observation,microscope observation and 16 S r RNA.A total of 182 aerobic/facultative anaerobes(148 strains in periodontal disease dogs and 34 strains in healthy dogs)and 119 anaerobes(96 strains in periodontal disease dogs and 23 strains in healthy dogs)were isolated and identified.Among the samples of healthy dogs,dogs with gingivitis and periodontitis,the highest proportion of aerobic/facultative anaerobic bacteria were Neisseria.The proportion of Neisseria in the affected dogs was as high as 37.16%(55/148),and Neisseria animalis was mainly(29/55,52.73%).The results of the isolation and identification of anaerobic bacteria show that the Fusobacterium sp.canine oral taxon has the highest isolation rate in both healthy dogs and dogs with gingivitis and periodontitis,which were 13.04%(3/23),13.33%(6/45),and 21.57%(11/51),respectively,and the separation rate gradually increased with the severity of the disease.The second anaerobic bacteria was Peptostreptococcus canis,the proportions of healthy dogs,gingivitis and periodontitis dogs were 8.70%(2/23),13.33%(6/45)and 17.65%(9/51),respectively.Neisseria are involved in early colonization and plaque formation.Because anaerobic bacteria are the direct pathogens causing PD,the high isolation rates of the Fusobacterium sp.canine oral taxon and Peptostreptococcus canis suggest that these two bacteria are important bacteria causing canine PD.4.The 16 S r DNA V3-V4 region of bacteria in subgingival samples of 5 healthy dogs,6 dogs with gingivitis and 5 dogs with periodontitis was amplified,and highthroughput sequencing technology was used to analyze the subgingival of dogs in different health states bacterial composition and diversity.The subgingival samples of PD dogs and healthy dogs both had higher homogeneity,but the bacterial richness of PD samples was significantly lower than that of the healthy group(p<0.05).It shows that the dominant bacteria of PD have inhibitory effect on general bacteria and break the bacterial balance.Bacteroidetes,Firmicutes,Proteobacteria,Fusobacterium,and Actinobacteria were the main subgingival bacteria in the healthy and diseased groups,and there was no difference at the phylum level.A total of 129 genera were obtained in all samples,and the highest relative abundance was Porphyromonas(34.25% in the healthy group,28.43% in the gingivitis group,and 25.67% in the periodontitis group).Secondly,the two genera with higher abundance in the healthy group were Fusobacterium(4.68%)and Cryobacter(3.78%),while those in the gingivitis group were Deinococcus(4.82%)and Fusobacterium(4.75%),Cryobacter(9.23%)and Fusobacterium(7.53%)in the periodontitis group.The relative abundance of the genera with relative abundance more than 1% in the three groups of samples showed that the Fusibacter and Peptococcus in the healthy group was significantly higher than that in the periodontitis group(p<0.05),Fusibacter and Deinococcus in the gingivitis group were significantly higher than those in the periodontitis group(p<0.05).The dominant bacterial genera in the healthy group were Porphyromonas,Leptotrichia,Catonella,Neisseria,etc.,the dominant bacterial genera in the gingivitis group are Actinobacteria,Peptostreptococcus,Deinococcus,Micrococcus,etc.,while the dominant bacteria in the periodontitis group are Prevotella,Coprococcus,Cryobacter,Filifactor,etc.Further analysis showed that the abundance of Porphyromonas and Filifactor in dogs with PD was significantly higher than that in healthy dogs.Prevotella melaninogenica and Prevotella Copri of the Prevotella are marker species for canine gingivitis and canine periodontitis,respectively.In summary,in this paper,the case analysis of canine PD was carried out,and the bacteria were isolated,cultured and identified from the subgingival samples of sick and healthy dogs,and the bacterial diversity of the subgingival samples was analyzed to explore the bacterial composition of canine PD.Its dominant bacteria and marker bacteria.the conclusions are as follows:(1)In this paper,a grading standard for canine PD was formulated based on the periodontal disease-related scoring standards,which can be used as the basis for the health status of the gums and their grading;the occurrence of canine PD is related to age,the severity of the disease increases with age.(2)Bacteroidetes and Firmicutes are the dominant bacterial phyla in canine PD,and the dominant bacterial genera for PD are Porphyromonas,Peptostreptococcus,Prevotella and Fusobacterium,and Prevotella is a marker species of canine PD.(3)Opportunistic pathogens such as facultative anaerobes such as Neisseria and Staphylococcus can colonize canine gums and form subgingival plaque.Anaerobic bacteria the Fusobacterium sp.canine oral taxon and Peptostreptococcus canis are the main pathogenic and dominant bacteria of canine PD. |
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