In performing phenotypic characterization tests, the organisms used for the inoculation of test media should be from fresh transfers and in good physiological condition. The methods described in this chapter are based primarily on methods developed for the characterization and identification of organisms which have usually been isolated on nutrient-rich media. While zoologists may be content to classify all insects or all vertebrates, prokaryote systematics generally deals with all prokaryotes and in an evolutionary context attempts to extend this study back to the origin of life itself. To test for acetamide hydrolysis, streak the surface of an acetamide agar slant with a sample from a dilute suspension. Incubate for up to 7 days. Care should be taken to ensure that the basal medium to which the carbohydrates are added does not contain fermentable or oxidizable sugars, and control media without any added carbohydrates should always be included during testing of cultures. Methanogenic members of the Archaea also fluoresce under UV light. Some facultative anaerobes, such as Escherichia coli and Staphylococcus aureus, grow aerobically by respiration with oxygen and grow anaerobically by fermentation.

Phenotypic Characterization and the Principles of Comparative Systematics

The fecal floras of polyp patients, Japanese-Hawaiians, North American Caucasians, rural native Japanese, and rural native Africans were compared. The polyp patients and Japanese-Hawaiians were considered to be groups at high risk of colon cancer, and the rural native Japanese and rural native Africans were considered to be groups at low risk. The North American Caucasians were found to have a flora composition intermediate between these two groups. Fifteen bacterial taxa from the human fecal flora were significantly associated with high risk of colon cancer, and five were significantly associated with low risk of colon cancer. Total concentrations of Bacteroides species and, surprisingly, Bifidobacterium species were generally positively associated with increased risk of colon cancer. Some Lactobacillus species and Eubacterium aerofaciens, which also produces major amounts of lactic acid, showed closest associations with low risk of colon cancer.

Intestinal floras of populations that have a high risk of colon cancer.

A total of 78 bacteriological samples were taken from the supragingival tooth surface after superficial cleaning with toothpicks or from the periodontal sulci of 42 affected sites in 21 adolescents or young adults with severe generalized periodontitis. Of 190 bacterial species, subspecies, or serotypes detected among 2,723 isolates, 11 species exceeded 1% of the subgingival flora and were most closely associated with the diseased sulci. Eleven others were also sufficiently frequent to be suspect agents of tissue destruction. Many of these species are known pathogens of other body sites. In addition, 10 species of Treponema were isolated. One of these and the "large treponeme" were also more closely associated with severe periodontitis than they were with healthy sites or gingivitis. There were highly significant differences between the composition of the flora of the affected sulci and the flora of (i) the adjacent supragingival tooth surface, (ii) the gingival crevice of periodontally healthy people, and (iii) sites with a gingival index score of 0 or 2 in experimental gingivitis studies. The floras of different individuals were also significantly different. There was no statistically detectable effect of sampling per se upon the composition of the flora of subsequent samples from the same sites. The composition of the supragingival flora of the patients with severe generalized periodontitis that had serum antibody to Actinobacillus actinomycetemcomitans was significantly different from the supragingival flora of patients without this serum antibody. However, there was no statistically significant difference in the composition of their subgingival floras.

/pdf/bacteriology-of-severe-periodontitis-in-young-adult-humans-3y14w11y5x.pdf

Bacteriology of severe periodontitis in young adult humans.

Statistical comparisons of the floras associated with juvenile periodontitis, severe periodontitis, and moderate periodontitis indicated that differences in the bacterial compositions of affected sites in these populations were not statistically significant. The subgingival flora of affected juvenile periodontitis sites was statistically significantly different from the adjacent supragingival flora and from the subgingival floras of people with healthy gingiva and of children with developing (experimental) gingivitis. However, the subgingival flora of affected juvenile periodontitis sites was not significantly different from the flora of sites with gingival index scores of 1 or 2 in adults with developing (experimental) gingivitis. Of 357 bacterial taxa among over 18,000 isolates, 54 non-treponemal species, 2 treponemal species, and mycoplasma were most associated with diseased periodontal sulci. These species comprised an increasing proportion of the flora during developing gingivitis and constituted over half of the cultivable flora of diseased sites.

https://iai.asm.org/content/iai/48/2/507.full.pdf

Comparative bacteriology of juvenile periodontitis.

A total of 171 taxa was represented among 1,900 bacterial isolates from 60 samples of sites affected with moderate periodontitis in 22 mature adult humans. The composition of the subgingival sulcus flora was statistically significantly different from that of the adjacent supragingival flora and the subgingival flora of 14 people with healthy gingiva, but was not significantly different from that of sulci affected with severe periodontitis in 21 young human adults. The sulcus floras of moderate periodontitis and severe periodontitis shared many of their predominant bacterial species, but there were differences in the relative proportions of some of these species. Similar relationships were found for seven taxa of treponemes that were cultured from the samples.

https://iai.asm.org/content/iai/42/2/510.full.pdf

Bacteriology of moderate (chronic) periodontitis in mature adult humans.

A polyacrylamide slab gel electrophoresis procedure was used to compare cellular proteins from bacterial isolates of gingival crevice floras. Isolates with identical protein patterns consistently were shown to be members of the same species. When used to screen isolates, the procedure reduced total analytical time and expense without sacrificing accuracy, and it provided additional verification of the identity of strains characterized by conventional phenotypic tests. Images

Polyacrylamide Slab Gel Electrophoresis of Soluble Proteins for Studies of Bacterial Floras

The soluble cellular proteins of strains of Peptococcus magnus and Peptostreptococcus micros were examined by polyacrylamide gel electrophoresis. The protein patterns were distinctive and repeatable, and the two species could be separated redily. The protein pattern of strain ATCC 23195, the type strain of Peptococcus glycinophilus (Cardon and Barker 1946) Douglas 1957, was identical to that of strain ATCC 33270 (= VPI 5464), the type strain of Peptostreptococcus micros (Prevot 1933) Smith 1957. Because these two strains were 84% homologous as determined by deoxyribonucleic acid-deoxyribonucleic acid homology experiments, the name Peptococcus glycinophilus is a later subjective synonym of Peptostreptococcus micros. Preliminary results indicated that ”Peptococcus variabilis“ (Foubert and Douglas 1948) Douglas 1957 may be a valid species, but reinstatement is not proposed at this time.

Synonomy of Peptococcus glycinophilus (Cardon and Barker 1946) Douglas 1957 with Peptostreptococcus micros (Prévot 1933) Smith 1957 and Electrophoretic Differentiation of Peptostreptococcus micros from Peptococcus magnus (Prévot 1933) Holdeman and Moore 1972

The soluble cellular proteins of strains of Peptococcus magnus and Peptostreptococcus micros were examined by polyacrylamide gel electrophoresis. The protein patterns were distinctive and repeatable, and the two species could be separated readily. The protein pattern of strain ATCC 23195, the type strain of Peptococcus glycinophilus (Cardon and Barker 1946) Douglas 1957, was identical to that of strain ATCC 33270 (= VPI 5464), the type strain of Peptostreptococcus micros (Prevot 1933) Smith 1957. Because these two strains were 84% homologous as determined by deoxyribonucleic acid-deoxyribonucleic acid homology experiments, the name Peptococcus glycinophilus is a later subjective synonym of Peptostreptococcus micros. Preliminary results indicated that “Pep tococcus variabilis” (Foubert and Douglas 1948) Douglas 1957 may be a valid species, but reinstatement is not proposed at this time.

Synonomy of Peptococcus glycinophilus (Cardon and Barker 1946) Douglas 1957 with Peptostreptococcus micros (PrCvot 1933) Smith 1957 and Electrophoretic Differentiation of Peptostreptococcus micros from Peptococcus magnus (PrCvot

D. E. Hash

Papers

Polyacrylamide Slab Gel Electrophoresis of Soluble Proteins for Studies of Bacterial Floras

Synonomy of Peptococcus glycinophilus (Cardon and Barker 1946) Douglas 1957 with Peptostreptococcus micros (Prévot 1933) Smith 1957 and Electrophoretic Differentiation of Peptostreptococcus micros from Peptococcus magnus (Prévot 1933) Holdeman and Moore 1972

Synonomy of Peptococcus glycinophilus (Cardon and Barker 1946) Douglas 1957 with Peptostreptococcus micros (PrCvot 1933) Smith 1957 and Electrophoretic Differentiation of Peptostreptococcus micros from Peptococcus magnus (PrCvot