Growth of Porphyromonas gingivalis, Treponema denticola, T. pectinovorum, T. socranskii, and T. vincentii in a chemically defined medium

Abstract

A chemically defined medium, OMIZ (Oral Microbiology and Immunology, Zurich)-W1 was developed. Medium OMIZ-W1 supports the long-term proliferation of a wide range of oral anaerobes, including representative strains of four Treponema species and Porphyromonas gingivaIss. High concentrations of ascorbic acid and ammonium ions proved to be important for the growth of these organisms. T. denticola CD-1 grew in the absence of polyamines and long-chain fatty acids, T. pectinovorum and T. socranskii required polyamines, whereas T. vincentii depended on both polyamines and lecithin for growth. Specific requirements for purines and/or pyrimidines were detected, and these requirements couid be used to distinguish Haemophilus-Actinobacilus group organisms. Some strains of P. gingivalis grew without vitamin K, while others were not satisfied by menadione but required its precursor 1,4-dihydroxy-2-naphthoic acid. Protoporphyrin IX or hemin equally satisfied the porphyrin requirements ofP. gingivalis and Bacteroidesforsythus, whereas ferrous sulfate was more efficiently used as a source of iron than was hemin. The cellular cohesiveness of P. gingivalis increased with high concentrations of hemin in the growth medium. PrevoteUa intermedia, B. forsythus, and several strains of P. gingivalis were more fastidious and required a protein or serum supplement to grow in medium OMIZ-W1.

Aspartame as a source of essential phenylalanine for the growth of oral anaerobes

“…The chemically defined medium OMIZ-Wl contains all common amino acids at millimolar concentrations [4]. It is therefore not surprising that growth of the 17 tested strains of oral anaerobes was not affected by the elimination of aspartic acid (Table 1), which can be readily formed from asparagine and glutamic acid.…”

“…The bacterial strains used in this investigation are listed in Table 1. Their origin and cultivation in medium OMIZ-Wl has been described previously [4], with the exception of the non-motile mutants CDK of Treponema denticola CD-1, and 33K of Treponema pectinovorum ATCC 33768.…”

“…These non-motile mutants showed the same response to amino acids and aspartame as the parental strains but, because of their colonial rather than diffuse growth in soft agar, differences were more readily evaluated. Tests were performed in 24-well multidishes (Nunc, Roskilde, Denmark) in soft agar medium OMIZ-Wl (prepared without aspartic acid and phenylalanine); amino acids and/or aspartame (98% purity, Bachem AG; Bubendorf, BL, Switzerland) were added to appropriate wells, and cells suspended in soft agar medium were added and cultured as described [4]. For Bacteroides forsythus FDC 331, which requires a peptide supplement to medium OMIZ-Wl for growth, tests were performed in the presence of both 1 mg/l bovine insulin and 0.1 mM L-threonyl-L-valine (Bachem AG).…”

“…Specific effects of defined nutrients on bacterial pro-256 liferation and community structure are not readily studied in vivo or in the complex media routinely used for in vitro cultivation. However, with the development of the chemically-defined medium OMIZ-Wl [4] it became possible to control the availability of phenylalanine and determined its role for the proliferation of oral anaerobes in vitro.…”

Outer sheath associated proteins of the oral spirochete Treponema maltophilum

“…The following strains were grown anaerobically in OMIZ-PAT medium as described previously [18]: Treponema vincentii LA-1 (ATCC 35580); T. vincentii Ritz A; Treponema denticola CD-1, ATCC 33521, ATCC 35405 T (T = type strain); Treponema pectinovorum ATCC 33768 T Escherichia coli DH5K was used for propagation of recombinant plasmid DNA. Plasmid vector pUC19 (Stratagene, Heidelberg, Germany) was the vector for routine subcloning and DNA sequencing.…”