Previous studies have demonstrated a direct iron-irreversible inhibition of a variety of microorganisms by human apolactoferrin. The present study compared the bactericidal effects of lactoferrin on Streptococcus mutans with the bacteriostatic effects of iron deprivation. Growth (as determined by change in optical density) and macromolecular synthesis, as determined by incorporation of 14Clabeled uracil, thymidine, and lysine, were inhibited by incubation of washed exponential-phase S. mutans NCTC 10449 with purified human apolactoferrin. Similarly, apolactoferrin inhibited glucose uptake and metabolism. Iron-saturated lactoferrin had no effect on bacterial growth or metabolism and was capable of serving as a source of iron in iron-depleted medium. S. mutans failed to grow, and there was no indication of macromolecular synthesis in iron-depleted partially defined medium; however, glucose metabolism continued, though at a reduced rate, and viability was retained for 72 h. There was no detectable metabolism of glucose by cells maintained for 18 h in iron-free medium. Metabolism was restored by transfer of iron-depleted S. mutans to iron-complete medium. This was in contrast to the irreversible inhibition by lactoferrin after 1 h of incubation. Inhibition could not be reversed by removal of cell surface-associated lactoferrin as detected by rhodamine isothiocyanate-labeled antilactoferrin. This inhibition of metabolism and rapid loss in viability observed with lactoferrin treatment suggest that lactoferrin has a direct bactericidal effect on S. mutans that cannot be attributed to simple iron deprivation.
E. coli O157:H7 is a pathogen that can be present in sewage contaminated waters. This organism poses a health risk for humans who come in contact with these waters via drinking, swimming, or shellfish consumption. A risk assessment model is needed to evaluate or quantify this risk. One possibility is the use of a computer model to simulate the fate and transport of E. coli O157:H7 downstream from a discharge point [e.g., a separate sanitary sewer overflow (SSO)]. However, this computer model would require input data regarding characteristics of this organism, which have not been previously available. One necessary input parameter is the rate at which die off of this organism occurs in a stream or river environment. Several studies were conducted to evaluate the die‐off rate of E. coli O157:H7 in an SSO impacted stream. Indicator bacteria (total coliforms, E. coli, and enterococci) were evaluated simultaneously. The results suggest that E. coli O157:H7 is not persistent — decay rates are high relative to the indicator bacteria. However, the decay plots suggest a biphasic response: initial decay is rapid, followed by an attenuated, slower decay. Hence traditional simulation methods using a single, first‐order decay rate may be inaccurate.
Lactoferrin is an iron-binding protein that is bactericidal against Streptococcus mutans and several other microorganisms. In this study, the influence of several physical conditions as well as the metabolic state of S. mutans on lactoferrin susceptibility were investigated. After exposure to lactoferrin, a 15-min lag period occurred before the initiation of killing, indicating that a two-step process is involved in lactoferrin killing. Cultures harvested during the early exponential phase were very sensitive to lactoferrin, whereas cultures harvested in the early stationary phase were markedly more resistant. The rate of killing was dependent on temperature; there was no loss of viability at 2 degrees C. Killing occurred at pH 5.0 to 6.0 in water and 20 mM glycine, but did not occur at any pH in 50 mM sodium phosphate or N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid (HEPES) buffer. Addition of exogenous ferrous or ferric ions did not reverse or prevent lactoferrin killing, nor did addition of 1 mM magnesium chloride.
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