Microtubules, undulipodia and Pillotina spirochetes.
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A prototype vaccine that is being developed for the control of swine dysentery (SD) was tested in two groups of experimental pigs. Vaccination induced high circulating antibody titres against the aetiological agent, Serpulina (Treponema) hyodysenteriae. Pigs in the first trial were vaccinated twice before being challenged orally with the bacteria. Five of 6 unvaccinated animals developed dysentery within a fortnight of challenge, but only 1 of 6 vaccinated pigs showed signs of disease at this time. Unexpectedly, 1 mo after challenge, the surviving unvaccinated pig and 2 remaining healthy vaccinated animals succumbed to the disease. The reason for the development of this late-onset form of dysentery was not clear. In the second trial, 8 pigs were vaccinated 3 times. Only 2 of these animals (25%) developed severe dysentery after being mixed with infected pigs, whereas 7 of 8 (88%) unvaccinated control pigs in the same pen became diseased. The late-onset form of dysentery was not observed. The prototype vaccine for SD provided a useful level of protection, and could be used in programs to control the disease in Australia.
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Ninety-five patients with periodontal disease each had subgingival plaque samples collected from four sites (one from each quadrant) in their mouths. The relative proportions of spirochaetes, motile rods and cocci were determined using dark field microscopy and the proportion of anaerobic to aerobic micro-organisms calculated after culture. In addition, clinical recordings were made. The only significant correlations observed were between probing depth or attachment loss and the proportion of cocci in the plaque (negative association), probing depth or attachment loss and sites which were suppurating or displayed a radiolucent interdental crest (positive association), and the percentage of cocci and sites with a radiolucent interdental crest (negative association). Partial correlation analysis controlling for probing depth or attachment loss showed that a significant inverse association between the percentages of cocci and the presence of a radiolucent interdental crest remained. No significant associations were observed between clinical signs such as bleeding on probing or suppuration and the microbiological assays. Overall there was a poor correlation between many of the signs thought by some to indicate periodontal disease activity.
Thirty-four patients with periodontal disease each had subgingival plaque samples collected from four sites (one from each quadrant) in their mouths. The relative proportions of spirochaetes, motile rods and cocci were determined using dark field microscopy and the proportion of anaerobic to aerobic microorganisms was calculated after culture. In addition, clinical recordings were made at these sampled sites. The patients then underwent a course of periodontal treatment and were placed on a maintenance programme. The clinical recordings were repeated and the results examined to ascertain if the original microbiological or clinical measurements could have been used to predict the response to therapy. Of the baseline recordings, the initial probing depth, the initial attachment level and the presence of suppuration all showed a positive correlation with the degree of pocket reduction or attachment gain produced by treatment. The percentage of cocci in the subgingival plaque correlated negatively with the treatment response. Suppuration seemed to be associated primarily with the original pocket depth while the percentage of cocci in subgingival plaque showed a true relationship with the amount of attachment gained after periodontal therapy. The significance of this finding is discussed.
BACKGROUND: Details of the development of human subgingival biofilm are unknown due to the difficulties in conducting experiments and especially in obtaining undisturbed materials. METHODS: This study was performed using deposits on carbonate apatite that had been inserted into human periodontal pockets for up to three weeks. Scanning electron microscopy using the vertically sectioned method and transmission electron microscopy using the freeze-substitution method were adopted. RESULTS: The development of subgingival biofilm occurred in five sequential phases: pellicle formation, microbial adherence, initial colonization, microbial organization, and establishment. Certain species in each of the initial, secondary and tertiary colonizers were considered to have a predilection for biofilm formation. Gram-positive, bacillary initial colonizers and gram-negative, filamentous secondary colonizers organized one stable structure that served as the framework for biofilm formation, and gram-negative, rod-shaped tertiary colonizers with cell-surface vesicles showed multigeneric coaggregation. The microbiota in the tertiary colonizers underwent repeated microflora alteration. CONCLUSIONS: Subgingival biofilm is constituted by initial, secondary and tertiary colonizers. Microflora alteration which is suggested to be related to periodontal disease, frequently occurred in the tertiary colonizers.
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Gene inventory and metagenomic techniques have allowed rapid exploration of bacterial diversity and the potential physiologies present within microbial communities. However, it remains nontrivial to discover the identities of environmental bacteria carrying two or more genes of interest. We have used microfluidic digital polymerase chain reaction (PCR) to amplify and analyze multiple, different genes obtained from single bacterial cells harvested from nature. A gene encoding a key enzyme involved in the mutualistic symbiosis occurring between termites and their gut microbiota was used as an experimental hook to discover the previously unknown ribosomal RNA-based species identity of several symbionts. The ability to systematically identify bacteria carrying a particular gene and to link any two or more genes of interest to single species residing in complex ecosystems opens up new opportunities for research on the environment.
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Three Millipore filters (100, 50, and 10 mmicro) can effectively and easily be used to isolate oral spirochetes from a mixed oral inoculum. The spirochetes appear pure in the underlying agar, owing to their ability to grow through the filter. Their passage through the 10-mmicro filters may be attributed to the instability of this pore size on storage.