Hepatozoon clamatae

Ferguson, Laura V., Hillier, N. Kirk & Smith, Todd G., 2013, Influence of Hepatozoon parasites on host-seeking and host-choice behaviour of the mosquitoes Culex territans and Culex pipiens, International Journal for Parasitology: Parasites and Wildlife 2 (1), pp. 69-76 : 70

2.3. Inoculation of mosquitoes and frogs with H. clamatae

To infect mosquitoes that would later be fed to frogs, two green frogs naturally infected with H. clamatae were placed in a 30 × 30 × 30 cm Plexiglas feeding cage containing 20 adult C. territans . Frogs were placed in the cage at dusk and left for three hours. Blood-fed mosquitoes, identified by their swollen, red abdomens, were removed from the feeding cage, placed in a holding container and provided with water and 10% sucrose. Mosquitoes were maintained for 30 days until parasites were mature. Six metamorph green frogs, three males and three females, were inoculated by force-feeding with infected mosquitoes, washed down with phosphate buffered saline ( PBS) (8.0 g NaCl/L, 0.20 g KH 2 PO 4 /L, 1.44 g Na 2 HPO 4 /L and 0.24 g KCl/L, pH to 7.4). Blood was obtained from each of the remaining frogs 35 days post inoculation and examined for H. clamatae in the same manner as previously described. Three out of six frogs became infected and were used in interrupted paired-choice and uninterrupted trials .