1- Fish for in vivo treatment:
A total number of 50 sea bream, from a naturally infested farm with protozoan Trichodina sp. and monogenetic trematode Furnestinia with an average body weight of 150 ±20 gm and an average length of 14 ±2 cm, were collected from a private fish farm in Diba triangle in Damietta Governorate. Fish were acclimatized to laboratory conditions at a water temperature 25 ±1 C º for one week before launching the experimental infection.
Clinical and post-mortem examination:
External examination of fish samples were performed to record clinical abnormalities and post-mortem changes according to (Noga, 2010)
Parasitological examination and identification:
Fish were examined externally with naked eye and with aid of magnifying glass for detection of parasitism according to (Noga, 1996). Mucous smears were immediately prepared from the skin and fins with the aid of microscopic slides and covers then examined with stereo microscope at 10 ,40 and 100 X magnification for detection of external parasites .Fish were euthanized and gills were carefully removed and placed in separate petri dish containing sterilized marine water to remove any excess gill mucus, and they were examined for parasites under a stereo microscope. All detected parasites; were collected, examined, fixed and stained according to (Lucky, 1977, Paperna and Laurencin, 1979) then identified according to (Lom, 1995, Paperna, 1996)
Vaccium210 is a commercial preparation of herbal extracts origanum oil (Carvacrol and Thymol) and Capsicum oil, produced by Slant Kim Company, Germany.
Experiment 1 :In vitro effect of Vaccium210 on Trichodina sp.
The experiment was designed according to (Ling et al., 2012, Fridman et al., 2014) as Infested Fish with Trichodina sp. were anaesthetised in 0.025% clove oil (Kildea et al., 2004). Mucus scraps from body surface were collected in small petri dish that was examined under stereo microscope to confirm parasitic existence and parasite livability. Afterwards, 20 ±1 parasites were transferred in each well of 96 well plate containing 0.5 mL filtered marine water from same fish source filtered through a filter with a pore size of 100 parasites /microscopic field.