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ABSTRACT. Porphyromonas gulae, a gram-negative black-pigmented anaerobe, is a pathogen for periodontitis in dogs. An approximately 41-kDa fimbrial subunit protein (FimA) encoded by fimA is regarded as associated with periodontitis. In the present study, the fimA genes of 17 P. gulae strains were sequenced, and classified into two major types. The generation of phylogenetic trees based on the deduced amino acid sequence of FimA of P. gulae strains along with sequences from several strains of Porphyromonas gingivalis, a major cause of human periodontitis, revealed that the two types of FimA (types A and B) of P. gulae were similar to type I FimA and types II and III FimA of P. gingivalis, respectively. A PCR system for classification was established based on differences in the nucleotide sequences of the fimA genes. Analysis of 115 P. gulae-positive oral swab specimens from dogs revealed that 42.6%, 22.6%, and 26.1% of them contained type A, type B, and both type A and B fimA genes, respectively. Experiments with a mouse abscess model demonstrated that the strains with type B fimA caused significantly greater systemic inflammation than those with type A. These results suggest that the FimA proteins of P. gulae are diverse with two major types and that strains with type B fimA could be more virulent. Periodontal diseases are characterized by gingivitis, a reversible condition involving gingival inflammation, and periodontitis, the irreversible destruction of periodontal tissues such as cementum, periodontal ligaments, and supportive bone [15]. In dogs, the prevalence of gingivitis is 95-100% and that of periodontitis, 50-70%, and the severity of periodontal disease tends to worsen with age, as in humans [8,10].Numerous periodontitis-related bacterial species have been reported in humans, with Porphyromonas gingivalis, a gram-negative black-pigmented anaerobe, considered a major species [15]. The 41-kDa cell surface protein fimbrillin (FimA), a subunit of fimbriae, is an important virulence factor of periodontitis, associated with adhesion to and invasion of gingival cells [4]. There are six genotypes of fimA encoding FimA (I through V and Ib), among which types II/ IV/Ib and I/III/V are considered invasive and non-invasive, respectively [2,3,[11][12][13][14]17].A previous analysis of 11 periodontitis-related bacterial species in oral swab specimens collected from 26 pet dogs in Japan showed that the isolation frequency of P. gingivalis was extremely low in dogs [9]. In contrast, Porphyromonas gulae was frequently detected in these specimens, accounting for approximately 70% of all Porphyromonas isolates. A recent study revealed the presence of a 41-kDa FimA, similar to that in P. gingivalis, in P. gulae [7]. In the present study, we performed molecular analyses of fimA in many clinical strains of P. gulae focusing on the association between the gene's diversity and virulence. Table 1 lists the P. gulae strains analyzed in the present study, 10 of which were isolated in Tokyo, Japan, previously [9]. The 7 new clinic...

ABSTRACT. We analyzed the distribution of 11 periodontitis-related bacterial species in dental plaque collected from 176 Japanese dogs divided into young (less than 2 years of age), middle-aged (2-7 years of age) and elderly (more than 8 years of age) groups using a polymerase chain reaction method. Clinical examination revealed that no dogs in the young group were affected by periodontitis, whereas the rates for gingivitis and periodontitis were high in the middle-aged and elderly groups. In addition, the total numbers of bacterial species in the middle-aged and elderly groups were significantly greater than in the young group. Our findings suggest that age is an important factor associated with the distribution of periodontitis-related bacteria and periodontal conditions in dogs. KEY WORDS: age, canine, dental plaque, periodontitis, periodontitis-related bacteria.doi: 10.1292/jvms.13-0041; J. Vet. Med. Sci. 75 (7): [999][1000][1001] 2013 Periodontal diseases are quite commonly identified in dogs. The diseases are composed of gingivitis, a reversible condition that features limited inflammation of gingival tissue, and periodontitis, which indicates destruction of periodontal tissues, such as the cementum, periodontal ligaments and supportive bone [8]. It has been summarized that the distribution frequency of gingivitis in dogs aged from 0 to 14 years old (average; approximately 6 years old) ranges from 95-100%, while that of periodontitis is from 50-70% [3].Our previous study analyzed the distribution of 10 human periodontitis-related species in dogs and found that specific species, such as Tannerella forsythia and Campylobacter rectus, were frequently detected [5]. In addition, Porphyromonas gulae, a major pathogen of periodontitis in dogs [2], was also detected at a high frequency [5]. On the other hand, the detection rates of other species, including Porphyromonas gingivalis, Treponema denticola, Capnocytophaga ochracea, Capnocytophaga sputigena, Prevotella intermedia, Prevotella nigrescens, Aggregatibacter actinomycetemcomitans and Eikenella corrodens, were quite low [5]. In addition, several periodontopathic species were shown to be possibly transmitted between dogs and their owners, although the distribution of periodontopathic species in both is generally different [12].In the present study, to investigate the influence of the age of dogs on the periodontal conditions and the distribution of the periodontitis-related species, 176 Japanese dogs were analyzed. On the basis of the age classifications of young (less than 2 years of age), middle-aged (2-7 years of age) and elderly (more than 8 years of age) dogs [6], the 176 dogs were classified into the young (n=39), middle-aged (n=62) and elderly (n=75) groups. The protocols used in this study were approved by

Two cDNA clones for human placental aromatase P-450 (P-450 AROM) have been isolated and sequenced. The insert of one clone (2894 bp) contains an open reading frame encoding a protein consisting of 503 amino acid residues together with a 49 bp S-untranslated stretch and a 1336 bp 3'-noncoding region to which a poly(A) tract is attached. Three potential poly(A) addition signals are detected in this 3'noncoding region. The other clone contains a shorter cDNA insert, the nucleotide sequence of which overlaps with most of the sequence of the longer cDNA insert (nucleotides 36-2355) except for one nucleotide substitution. The 3'noncoding region of this shorter cDNA is only 846 bp in length, but a poly(A) tract is also attached to its 3'-terminus. Northern blot analysis of human placental RNA reveals the presence of two major mRNA species of 3.4 and 2.9 kb when probes excised from the overlapping region of these two cDNAs are employed. The 2.9 kb mRNA is not detected, however, when a fragment of the non-overlapping region of the longer cDNA is used as a probe. It is therefore concluded that the two major species of P-4SOAROM mRNA are formed as a consequence of alternative processing of precursor mRNA(s).

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