Characterization of black-pigmented Bacteroides strains isolated from animals

The aims of the study were: the isolation of strains of black-pigmented Bacteroides from the gingival sulcus of different animals, their biochemical and immunological characterization and comparison of their properties for classification within the genus. A total of 104 strains, isolated from cats, dogs, racoons and a jaguar, were characterized on the basis of fermentation of carbohydrates, metabolic end products, haemagglutination studies, enzymatic activities, catalase production and indirect immunofluorescence. No differences were observed between the strains regardless of their animal origin. The strains did not ferment carbohydrates, produce phenylacetic acid, show an array of enzyme activities or agglutinate sheep red blood cells. They were catalase-positive and so differed from the human oral strains of Bact. gingivalis. Immunofluorescence microscopy revealed that the animal strains shared at least one major antigen with Bact. gingivalis but none with Bact. asaccharolyticus. Apart from their catalase activity, the animal strains isolated were similar to those of human Bact. gingivalis strains.     

Characterization of black-pigmented Bacteroides strains isolated from animals

The aims of the study were: the isolation of strains of black-pigmented Bacteroides from the gingival sulcus of different animals, their biochemical and immunological characterization and comparison of their properties for classification within the genus. A total of 104 strains, isolated from cats, dogs, racoons and a jaguar, were characterized on the basis of fermentation of carbohydrates, metabolic end products, haemagglutination studies, enzymatic activities, catalase production and indirect immunofluorescence. No differences were observed between the strains regardless of their animal origin. The strains did not ferment carbohydrates, produce phenylacetic acid, show an array of enzyme activities or agglutinate sheep red blood cells. They were catalase-positive and so differed from the human oral strains of Bact. gingivalis. Immunofluorescence microscopy revealed that the animal strains shared at least one major antigen with Bact. gingivalis but none with Bact. asaccharolyticus. Apart from their catalase activity, the animal strains isolated were similar to those of human Bact. gingivalis strains.     

Characterisation of Bacteroides from sheep periodontal disease by SDS-PAGE of outer membrane proteins

Isolates of Bacteroides species obtained from a longitudinal study of developing periodontal disease in sheep were analysed by SDS-PAGE. Protein profiles of Sarkosyl-insoluble outer membrane extracts were compared within groups of isolates which had already been defined by conventional biochemical techniques. Heterogeneity was exhibited within most groups. Isolates of B. gingivalis and B. asaccharolyticus shown to be similar to human isolates by conventional biochemical tests, gave different protein profiles from the respective type cultures. The sheep B. gingivalis-like isolates were however homogeneous, while the B. asaccharolyticus-like organisms could be divided into 3 subgroups. SDS-PAGE appears to be a useful tool for the examination of bacterial flora and recognition of subgroups of subspecies.     

Enzymatic characterization of oral and non-oral black-pigmented Bacteroides species

The purpose of this study was to investigate the production of various enzymes by oral and non-oral black-pigmented Bacteroides species using chromogenic substrates. The 19 substrates present in the API ZYM system did not differentiate between B. melaninogenicus, B. denticola, B. loescheii and B. levii. The asaccharolytic black-pigmented Bacteroides species showed each species specific enzyme activity, however, differences were based on one enzyme only as far as B. asaccharolyticus and B. endodontalis are concerned. An extended number of 40 chromogenic substrates were tested in order to find more species specific enzyme. With a set of 20 substrates it appeared to be possible to discriminate between all species tested. The possibility to use enzymes for the identification of black-pigmented Bacteroides is discussed.     

Properties of oral asaccharolytic black-pigmented Bacteroides

Bacteroides endodontalis, a newly described asaccharolytic black-pigmented Bacteroides, along with the other two recognized species of this group (B. gingivalis and B. asaccharolyticus) were studied for their susceptibility to various dyes and inhibitory agents and for some of their enzymatic activities to facilitate differentiating between them. Bacteroides endodontalis resembles B. asaccharolyticus physiologically except for the fact that the former cannot grow on media containing methylene blue, neutral red, or 3% sodium chloride, whereas B. asaccharolyticus can. On the other hand, B. endodontalis and B. gingivalis can grow on a medium containing Congo red while B. asaccharolyticus cannot.     

Proteolytic activity of black-pigmented Bacteroides strains

Abstract The proteolytic activity of several black-pigmented Bacteroides species was measured. Bacteroides gingivalis was the only species having collagenolytic activity. General proteolytic activity on gelatin and Azocoll was shown in cultures of B. gingivalis, B. asaccharolyticus, B. endodontalis, B. intermedius, B. corporis and to a lesser extent B. melaninogenicus; B. loescheii did not show proteolytic activity. When culture filtrates were tested, B. gingivalis showed high cell free proteolytic activity, whereas the other species had only very weak cell free activity. Growth curves of B. gingivalis revealed two distinct proteolytic activities; general proteolytic activity was found during the logarithmic growth phase, whereas a second peak containing high collagenolytic activity was found after prolonged incubation of cells showing autolysis.     

Oxygen tolerance of oral and non-oral black-pigmented Bacteroides species

Abstract The oxygen tolerance of 24 black-pigmented Bacteroides (BPB) strains, including 9 species, was investigated. For this study the survival of single bacterial cells on blood agar medium was determined. It was found that BPB species are not equally tolerant to atmospheric oxygen at room temperature. Among the asaccharolytic species, B. endodontalis showed the highest oxygen sensitivity, while B. gingivalis and B. asaccharolyticus survived for more than 24 h under atmospheric conditions. All saccharolytic species survived at least 4 h of exposure to air.     

Intrafamilial transmission of black-pigmented, putative periodontal pathogens

Porphyromonas gingivalis and Prevotella intermedia are black-pigmented, putative periodontopathogenic bacteria considered to cause some forms of periodontal disease. Porphyromonas gingivalis and P. intermedia can be transmitted between humans and produce periodontal disease in susceptible hosts. In this article, studies using molecular typing methods for determining the transmission of black-pigmented, putative periodontopathogens between family members are reviewed. As individuals living close to each other are more prone to transmit bacteria, the studies on transmission of periodontopathogens have been performed on family members. It has been shown that black-pigmented bacteria are not only transferred between spouses but also between parents and child. Since only a limited number of studies have been done, longitudinal and controlled studies should be carried out to elucidate further the transmittance potential of these bacteria.     

Serological properties and immunobiological activities of lipopolysaccharides from black-pigmented and related oral Bacteroides species

Lipopolysaccharides (LPS) from five species of oral Bacteroides, B. gingivalis strains 381 and ATCC 33277, B. oralis ATCC 33269, B. loescheii ATCC 15930, B. intermedius ATCC 25611 and B. corporis ATCC 33547, were extracted from whole cells by the phenol/water procedure, and subsequently purified by treatment with nuclease and ultracentrifugation. The LPS were composed of hexoses, glucosamine, fatty acids and phosphorus. Heptose and 2-keto-3-deoxyoctonate were not detected. The LPS preparations from B. gingivalis strains 381 and ATCC 33277 presented very similar SDS-polyacrylamide gel electrophoresis patterns when stained with ammoniacal silver. They produced a fused precipitin band against an antiserum to B. gingivalis 381 LPS in immunodiffusion tests. Antisera raised against the LPS from B. loescheii and B. intermedius reacted with the LPS prepared from all the oral Bacteroides strains except those of B. gingivalis. All the LPS preparations were mitogenic for spleen cells of BALB/c (nu/nu) mice, but not for thymus cells from C3H/HeN mice. The LPS induced marked mitogenic responses and polyclonal B cell activation for spleen cells of not only C3H/HeN (LPS responder) mice, but also C3H/HeJ (LPS nonresponder) mice. The mitogenic responses were not suppressed significantly upon addition of polymyxin B to the reaction mixture. These LPS also enhanced interleukin-1 production by murine peritoneal macrophages and mouse cell line J744. 1 macrophages. Hydrolysis of B. gingivalis ATCC 33277 LPS in 1 m-HCl at 100 degrees C for 1 h yielded lipid and polysaccharide. The lipid portion was largely composed of fatty acids and glucosamine, and was mitogenic for spleen cells from C3H/HeJ as well as C3H/HeN mice, while the polysaccharide portion induced no significant mitogenic responses under similar experimental conditions.     

Detection of conjugal transfer systems in oral, black-pigmented Bacteroides spp.

Oral, black-pigmented Bacteroides spp. are important pathogens in oral anaerobic infections and dental disease. We detected conjugation systems in isolates of Bacteroides denticola and Bacteroides intermedius that transferred tetracycline resistance (Tetr) and penicillin resistance to Bacteroides buccae and to Bacteroides fragilis, an intestinal Bacteroides species. A cloned Tetr gene from B. fragilis hybridized to the transferable Tetr locus in the oral strains, indicating that genetic exchange occurs between these two groups of anaerobes.     

口腔各种产黑色素类杆菌外膜结构的电镜观察

本文对从根管中分离的产黑色素类杆菌群的部分菌株:牙髓类杆菌、牙龈类杆菌、中间型类杆菌、产黑色素类杆菌的表面结构进行了电镜观察。观察发现牙髓类杆菌、牙龈类杆菌表面有放射状排列的菌毛样结构;牙龈类杆菌表面有荚膜;多数细菌的细胞膜外有多个小泡状结构;这些结构可能在细菌吸附于组织的过程中起着重要的作用。     

Enterotoxin-producing Bacteroides fragilis strains isolated from horses]

Seven Bacteroides fragilis strains were cultured from samples collected from horses. From all the tested strains, as well as from the reference B. fragilis strains: enterotoxigenic NCTC 11925 and nonenterotoxigenic IPL 323 strain, DNA was isolated using Genomic DNA PREP PLUS isolation kit manufactured by A&A Biotechnology (Poland). To detect the enterotoxin (fragilysin) gene, polymerase chain reaction (PCR) was applied, using the following starters: 404 (GAG CCG AAG ACG GTG TAT GTG ATT TGT) and 407 (TGC TCA GCG CCC AGT ATA TGA CCT AGT). DNA obtained from bacterial cells was amplified in a thermocycler (Techne). The temperature profile was as follows: 1 cycle (4 min. 94 degrees C), 40 cycles (1 min. 94 degrees C, 1 min. 52 degrees C, 1 min. 74 degrees C). Amplification products were detected by electrophoresis in agarose gel (1%) with ethidium bromide added. The presence of the fragilysin gene was detected in two strains. Among the strains isolated from horses enterotoxin gene-possessing Bacteroides fragilis strains (ETBF) can be detected.