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Inhibition of Mastitic Bacteria by Bovine Milk Apo-Lactoferrin Evaluated by In Vitro Microassay of Bacterial Growth¹

National Animal Disease Center, USDA, Ames, IA 40010

K. Larry Smith

Department of Dairy Science, Ohio Agricultural Research and Development Center, The Ohio State University, Wooster 44691

ABSTRACT

An in vitro microassay was developed to evaluate antimicrobial properties of bovine apo-lactoferrin. The growth of coliform, staphylococcal, and streptococcal bacterial strains in a defined synthetic medium was inhibited by bovine apolactoferrin (.5 to 30.0 mg/ml). Addition of iron-saturated lactoferrin to the synthetic medium did not inhibit growth of test strains. Inhibition by apo-lactoferrin was greater for coliform than Gram-positive strains for all concentrations of apo-lactoferrin evaluated. No concentration of apo-lactoferrin proved bactericidal for either coliform or Gram-positive strains. Inhibition of two coliform strains by apo-lactoferrin (10 mg/ml) was abolished by addition of ferric iron to the assay system, indicating an iron-dependent nature of apo-lactoferrin induced inhibition of bacteria. Bicarbonate supplementation of the growth system containing apo-lactoferrin (1 mg/ml) increased inhibition of three coliform strains by apo-lactoferrin. Addition of increasing concentrations of citrate (2.0 mg/ml) to an assay system containing apo-lactoferrin (5 mg/ml) resulted in a concomitant reduction of growth inhibition of three coliform strains. These data indicate a potential relationship between the molar ratio of citrate to lactoferrin of the lacteal secretion and its capacity to inhibit coliform strains associated with mastitis.

¹ Salaries and research support provided by State and Federal Funds appropriated to the Ohio Agricultural Research and Development Center, The Ohio State University. Journal Article No. 70–83.

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