Neobenedenia sp.

Trujillo-Gonzalez ́, Alejandro, Constantinoiu, Constantin C., Rowe, Richard & Hutson, Kate S., 2015, Tracking transparent monogenean parasites on fish from infection to maturity, International Journal for Parasitology: Parasites and Wildlife 4 (3), pp. 316-322 : 317

2.1. Source of fi sh and Neobenedenia sp.

Fifty hatchery reared L. calcarifer (150 ± 30 LT mm) were maintained in 100 L fresh water aquaria at the Marine Parasitology Laboratory, James Cook University. Fish had not been previously exposed to Neobenedenia . Fish were acclimated to sea water 24 h prior to experiments by increasing salinity to 10, 20, 30 and 35 ppt over 2 h intervals. Fish were fed until satiation every two days (~ 1 g per fish) with pellets formulated for L. calcarifer (Ridley Aqua- Feed™). Parasite eggs were sourced from an experimental infection in the laboratory, which was established using methods previously described ( Militz et al., 2013). Neobenedenia sp. investigated in this study is presently unidentified given the absence of diagnostic criteria to differentiate between geographical/host isolates and species ( Whittington, 2004, 2012). Phylogenetic analysis of approximately 12 Neobenedenia spp. isolates collected from multiple fish hosts in northern Australia is ongoing and may provide species level-clarification (Brazenor, unpublished data). Meanwhile, representative specimens mounted on slides were accessioned in the South Australian Museum, Australia (SAMA) in the Australian Helminth Collection (AHC); SAMA AHC 35461 (see Hutson et al., 2012). Parasite eggs were collected daily and held in Petri dishes with fresh sea water. Newly hatched oncomiracidia (<3 h old) were gently aspirated with a pipette and used in the experiments described below.