The Influence of Hyaluronidase and Hyaluronidase Substrates on Penetration of Cultured Cells by Eimerian Sporozoites

SYNOPSIS Leighton tube cultures of bovine embryonic kidney cells were inoculated with Eimeria adenoeides sporozoites suspended in media containing either hyaluronidase, hyaluronidase substrates (chondroitin sulfate and hyaluronic acid) or Ficoll. After 1 hr at 41 C, coverslips were removed and cells were fixed and stained. Hyaluronidase (1 and 10 mg/ml) did not increase the number of intracellular sporozoites. Chondroitin sulfate (1 and 10 mg/ml) and hyaluronic acid (1 mg/ml) did not reduce the number of intracellular sporozoites. However, the number was reduced when the media contained either chondroitin sulfate (100 mg/ml) or hyaluronic acid (5 mg/ml), which were quite viscous.
Ficoll (117 mg/ml), which produced the same viscosity as 5 mg hyaluronic acid/ml, also reduced the number of intracellular sporozoites. This finding circumstantially indicates that sporozoites may be physically inhibited from entering cells by the high viscosity of the substrates.
Biochemical tests, which detected as little as 0.2 μg of known hyaluronidase, failed to detect hyaluronidase activity in excysted intact or fragmented E. adenoeides sporozoites or in sporozoites within E. tenella oocysts.