Antimicrobial activity in the egg wax of the African cattle tick Amblyomma hebraeum (Acari: Ixodidae)

Eggs of the tick Amblyomma hebraeum Koch (Acari: Ixodidae) inhibited the growth of Escherichia coli and Serratia marcescens (Gram-negative bacteria) in solid culture, but not the growth of Staphylococcus epidermidis, and only marginally the growth of Bacillus subtilis (Gram-positive bacteria). When egg wax was extracted with chloroform/methanol (2:1), the extract contained antibacterial activity, but the denuded eggs did not. When assayed against bacteria in liquid culture, the aqueous phase inhibited the growth of S. epidermidis. However, the activity against E. coli was lost during extraction. The antimicrobial component of the aqueous phase was heat stable (100 degrees C for 10 min), resistant to proteinase K (15 min at 55 degrees C) and to pronase (30 min at 37 degrees C). The antibacterial activity in the aqueous phase increased the permeability of the cell membrane of susceptible bacterial cells within 30 min. However, lysis of the cells was detected by optical density measurements (OD600 nm) only after 1.5 h. The most evident cytological changes observed by transmission electron microscopy were a thickening of the cell wall and the appearance of numerous electron lucent areas within the cytoplasm of treated bacteria. Gene's organ, the egg-waxing organ in ticks, grew enormously during the first 16 days post-engorgement, and gained antimicrobial activity by day 10 (when oviposition began). This suggests that Gene's organ is the major source of the antibacterial substance in the egg wax. The vitellogenic hormone in A. hebraeum, 20-hydroxyecdysone, when injected into recently engorged females, did not stimulate growth of Gene's organ or precocious secretion of antimicrobial activity.